Opportunities for Postdoctoral Fellows in Electron Cryomicroscopy and Structural Biology Prof. David DeRosier (derosier-at-brandeis.edu). Rosenstiel Basic Medical Sciences Research Center Brandeis University
I have a postdoctoral position to study the structure of large macromolecular machines. The position is fully funded for at least two years, and we will pay competitive salaries. Write me if you or someone you know is interested. Here is a brief description of the available projects:
The cytoskeleton: The actin cytoskeleton is a set of cellular machines responsible for many of the dynamic capabilities of eukaryotic cells. Actin bundles are a key elements used to shape cells, generate filopodia, and order the cell cytoplasm. The project is aimed at determining the structure of the actin bundle in the brush border cells, as a model system. These bundles are the best characterized and most tractable bundles. The approach is two pronged beginning with in vitro studies of the components (actin, fimbrin and villin) and leading to in vivo studies of intact bundles. The tool we use is electron cryomicroscopy and image analysis.
The bacterial flagellum: The flagellum is a several machines in one. It is composed of over a dozen different proteins, with one set making up the rotary motor, another the drive train, another a bushing and seal, and another the flagellar export apparatus. We determining the structures of these components using electron cryomicroscopy and image analysis, and we are correlating structural features with the primary sequences of the component proteins.
The bacterial signaling complex: The bacterial cell senses the environment and transmits a signal to the flagellum. We are using electron cryomicroscopy and image analysis to determine the structure of a 1.4 megadalton complex of the cytoplasmic domain of a chemoreceptor, a kinase, and an adaptor molecule. The complex is highly active enzymatically, but has an unusual stoichiometry, which suggests signal integration from several receptors into a single kinase.
Professor David J. DeRosier Abraham S. and Gertrude Burg Chair of Life Sciences W.M. Keck Institute for Cellular Visualization Rosenstiel Basic Medical Sciences Research Center MS029 Brandeis University 415 South Street Waltham, MA 02454-9110 Telephone: 781-736-2494 FAX: 781-736-2405 email: derosier-at-brandeis.edu
I need to find someone in the Washington, DC area (or close by) who repairs RMC ultramicrotomes. We have an MT-7 that has a very fine chatter visible only under the beam.
If you know of anyone/company that does this type or work, please let me know.
I don't mind chatter, just not on my sections.
Paula :-)
p.s. hey sectioner, sectioner.....Breathe!
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We are looking for a specimen rotating holder for a JEOL 100CX part# IEM100/200CX-SRH. Also for the JEOL 100CX we are in need of a dispensing magazine, receiving magazine and/or cassette film holders. If you are looking for a home for any of these items please contact me.
Thank you,
Ruth Yamawaki Department of Comparative Medicine Stanford University (650) 723-3457 FAX (650) 498-6259
Hello All I need to purchase a new 3mm / 5mm Diamond knife for ultrathin sections of calcified tissue and polyurethane samples in Spurs resin. The prices from most of the suppliers are horrendous, as you well know. Does anyone out there have any of these lying around which are not used? As a last resort we'll consider a good, used one.Any brand name would do. And where can I get my "old" one re-sharpened at a reasonable price? Thanks and regards Nazlia
I am looking for a high end deconvolution program that would also include image acquisition capabilities. I colleague would like to do both with a single software package.
Any suggestions would be appreciated. Thank you.
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8 Canada
Paula: For many years we have relied on two firms for ultramicrotome maintenance and repair work. Both firms we found could be counted on for prompt and excellent work. For our MT-series ultramicrotomes we have engaged the services of Bill McGee of Microtome Service Co. (315)-451-1404. For our Reichert and LKB instruments we relied on Helmut Ptzig and Mario Saccoccio of Microscopical Optical Consulting Inc. (914) 268-6450 or e-mail MOCLeica-at-aol.com.
Its possible that the chatter is not being generated from the ultramicrotome, but from some outside source of vibration. If so, you may want to install an anti-vibration platform. Regards, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences State University of New York-Binghamton Binghamton, NY 13902-6000
I have inherited a 1980s JEOL X-Ray Counting Unit Type XCU, which I want to press into service as a ratemeter for wds alignment purposes.
Is anyone out there sufficiently familiar with this model and sufficiently generous with their time to reply to me off-list so that I can be enlightened?
thanks
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Diamond knife manufacturers are mostly located in Switzerland and USA and they will tell you that prices have remained constant or decreased over the past 20 years. But you are right too, countries whose currencies have not kept pace with the US$, the Swiss Franc and the Euro find diamond knives and many other imported items very expensive indeed. The Australian dollar 2 years ago was equal to US 65 cents. Now it is 48.5 cents; a decline of 25% at a time of lower inflation here than in the USA. When our business was started 22 years ago we received US$1.20 for one A$. This topic is peripheral to the listserver, but the exchange rate woes do affect numerous microscopists. There is nothing we can do, but to encourage microscopists in the high exchange rate countries to come and visit our country. Australia is cheap for you to travel in, as our $ buys as much of most Australian made goods as the US$ buys in the USA. We have an excellent conference planned in February 2002 in Adelaide. Spend your money here and support our A$ ! Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, October 02, 2001 8:42 PM, Nazlia [SMTP:ctssamodien-at-samiot.uct.ac.za] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello All } I need to purchase a new 3mm / 5mm Diamond knife for ultrathin } sections of calcified tissue and polyurethane samples in Spurs resin. } The prices from most of the suppliers are horrendous, as you well } know. Does anyone out there have any of these lying around which are } not used? As a last resort we'll consider a good, used one.Any brand } name would do. And where can I get my "old" one re-sharpened at a } reasonable price? } Thanks and regards } Nazlia
I can't find compagnies which have spin-polarization detectors (magnetic microstructures in SEM) in their sales program. Does anyone out there can help.
Ing. Rudy De Vos K.U.Leuven - Department MTM Kasteelpark Arenberg 44 B - 3001 Heverlee Belgium
} Good Morning, } Does anyone out there know why you add ammonium chloride to a pre } incubating step? Caroline Miller ****************** It quenches fluorescence from unreacted aldehydes.
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
you are trying to bind up all the free aldehyde groups left over from fixation with aldehydes so they don't react with your antibodies. glycine works too. tom
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear listers, Could someone please send me the confocal istserve URL for subscription to that list. Many thanks, Jeremy jb_sanderson-at-yahoo.com
____________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk or your free -at-yahoo.ie address at http://mail.yahoo.ie
Anyone know of a good repair facility in PA or close by for a Bausch & Lomb Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads.
Dave Stadden Armstrong World Industries, Inc. 717-396-5109 drstadden-at-armstrong.com
Does anyone give me informations about Oxford INCA 100 EDS file format? Any reference for an INCA {--} EMS spectra converter?
P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
As has been pointed out, ammonium chloride reduces aldehyde double bonds so they won't react with your antibodies. Another advantage of reducing double bonds is to cut down the amount of autofluorescence in your specimen (in other words, places that fluoresce nonspecifically, because of double bonds). Ammonium chloride is relatively gentle in this regard. If your specimen has strong autofluorescence, you may need to resort to a more drastic procedure involving sodium borohydride (which is toxic, so has to be handled carefully). A good article that deals with sodium borohydride and bond reduction is:
Eldred et al, 1983, Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry, J Histochem Cytochem 31:285-292.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Is there a procedure for determining the shading of a fluorescence microscope image? I assume that one needs to take an image of a uniform object, but are there standard objects? Will a volume object work, or must I use a surface object. Would a front-surface mirror work, assuming I could measure the leakage through the barrier filter? Everett Ramer
To my understanding (which is admittedly weak in this area) the preferred method for visualizing membrane structure is to use freeze-fracture, then carbon replica, then TEM. Does anyone have an opinion as to whether high resolution FE-SEM of metal-coated samples would be as effective in this regard as viewing a replica in the TEM.
Bob
Robert R. Wise, Ph.D. University of Wisconsin-Oshkosh
Currently on sabbatical at UW-Madison Botany Department 430 Lincoln Drive Madison, WI 53706 email: wise-at-uwosh.edu phone: 608-262-4288 fax: 608-262-7509
TekNet has fixed several of our binocs. Email Jon Petz and give him your model info, etc., to see if he can work on it. He's in your area.
jon-at-teknetinc.com
Sara
On Wed, 3 Oct 2001 David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com wrote:
} Date: Wed, 3 Oct 2001 12:01:31 -0400 } From: David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com } To: Microscopy-at-sparc5.microscopy.com } Subject: East coast repair for B&L scope? } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } } } Anyone know of a good repair facility in PA or close by for a Bausch & Lomb } Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads. } } Dave Stadden } Armstrong World Industries, Inc. } 717-396-5109 } drstadden-at-armstrong.com } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Try the Leica Inc. facility in Buffalo NY and ask for Ken Brown. I think he can help you. The phone number is (716)686-3000 or if needed the main Leica # 1-800-248-0123. Leica owns B&L and handles parts for these microscopes.
Dave Joswiak Dept. of Astronomy University of Washington Seatle, WA 98195 (206)543-7702
On Wed, 3 Oct 2001 David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Anyone know of a good repair facility in PA or close by for a Bausch & Lomb } Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads. } } Dave Stadden } Armstrong World Industries, Inc. } 717-396-5109 } drstadden-at-armstrong.com } } } }
In INCA select the spectrum in any of the Navigator windows (Acquire, Confirm, Report etc.), right mouse click, then select 'Export'. You have the choice of image and file types inculding EMSA format.
Ron
On Wed, 3 Oct 2001 19:19:49 +0200 FABBRI {fabbri-at-cigssrv1.unimo.it} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anyone give me informations about Oxford INCA 100 EDS file format? } Any reference for an INCA {--} EMS spectra converter? } } P.L. Fabbri } } --------------------------------------------- } Dr. Pier Luigi Fabbri } C.I.G.S. - Centro Interdip. Grandi Strumenti } Università di Modena } via Campi 213/A - 41100 Modena, Italy } Tel +39-059-2055231 / +39-059-370551 } Fax +39-059-370551 } E-mail: fabbri-at-mail.cigs.unimo.it } --------------------------------------------- } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
We have been very happy with Paul Musci of M&R Optical. He is a former B&L serviceman. He can be reached at:
PO Box 515 Charlton City, Massachusetts 01508 508-248-4658
Helen Mayer UCAR Carbon Parma, OH
-----Original Message----- } From: "David_R_Stadden-at-armstrong.com"-at-sparc5.microscopy.com [mailto:"David_R_Stadden-at-armstrong.com"-at-sparc5.microscopy.com] Sent: Wednesday, October 03, 2001 12:02 PM To: Microscopy-at-sparc5.microscopy.com
Anyone know of a good repair facility in PA or close by for a Bausch & Lomb Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads.
Dave Stadden Armstrong World Industries, Inc. 717-396-5109 drstadden-at-armstrong.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We still have a few spots available for our Microwave Specimen preparation workshop. The information follows. Please let me know immediately if you would like to attend. Thanks, Debby
===========================
The workshop will be held October 21, 22, and 23 in the Life Science Microscopy Facility at Purdue University, Wst Lafayette, Indiana, and run by Richard Giberson of Ted Pella, Inc.
Sunday Oct 21: introductory session starting at 2:00pm Monday Oct 22: Hands-on prep for TEM and SEM Tuesday Oct 23: Hands-on prep for ICC including block prep and ICC protocol.
We will have two microwave ovens, 2 TEM's, 1 SEM and 4 microtomes available during this time so there should be plenty of opportunity for examining the final products.
Cost for the workshop will be $350. This registration cost will include the book " Microwave Techniques and Protocols" by Giberson and Demaree, meals from dinner on Oct 21 through lunch on Oct 23, and most materials. Participants will have to provide their own resins if using ones other than Epon generic, Spurr's, or LR White. Also participants will have to provide their own specimen material (call and we will try to obtain it here if possible) and diamond knives for microtoming. Check with us if you need any other special reagents to determine if we can provide them.
The workshop will be limited to 12 participants. Let me know immediately if you would like to register and I will send info on where to send the registration checks. Please note that once you have registered the registration costs can only be refunded if we can fill the spot. All fees will be refunded if the workshop must be cancelled for any reason.
Participants will also have to provide their own transportation and hotel rooms. We have a block of rooms reserved at the University Inn. The Purdue rate is $70 for 1 person and $77/room for 2 people. We will be happy to try to match up participants who would like to share a room. Contact me for reservation information.
Those who plan to drive in can contact me for instructions. West lafayette is reached via I-65 from Indianapolis (~1 1/4 hr northwest) or Chicago (2+ hr. southeast) and via I-74 from central Illinois (~1 1/2h northwest from Champagne-Urbana).
Those who plan to fly can reach us by flying into Indianapolis and taking ground transportation to campus (~ 1 1/2 hr.. and $20 one-way). Northwest Airlines has commuter flights directly into the Lafayette/Purdue airport from Detroit. Note that we stay on Eastern Standard time all year round so are on Central time in the summer and Eastern time in the winter (at least we don't have to remember to change our clocks!). Planes presently fly into Lafayette leaving Detroit at 5:10 Saturday nights. There are only evening flights coming in on Sunday. Flights to Detroit leave at 6:01 in the evening or 7:15 in the morning during the week.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
On Wednesday, September 26, 2001 8:48 AM, Tindall, Randy D. {TindallR-at-missouri.edu} wrote: } Hi Debby, } } I'd like to receive information about this workshop. (Lou Ross forwarded } your message to me.) I doubt if I can get funding to make it, but I may } try. } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } }
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA22214 for dist-Microscopy; Thu, 4 Oct 2001 10:30:25 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA22206 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 4 Oct 2001 10:29:54 -0500 (CDT) Received: from umc-mail01.missouri.edu (umc-mail01.missouri.edu [128.206.10.216]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA22199 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 4 Oct 2001 10:29:43 -0500 (CDT) Received: from [128.206.78.175] (mu-078175.dhcp.missouri.edu [128.206.78.175]) by umc-mail01.missouri.edu with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2653.13) id 4GNN209J; Thu, 4 Oct 2001 10:26:31 -0500 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Sender: RossLM-at-pop.email.missouri.edu Message-Id: {v04011700b7e21dd57438-at-[128.206.78.175]}
Hi,
The Microbeam Analysis Society (not MSA) membership renewal packets were mailed out late last week. Inside the packet you will find a membership renewal form, a M&M Journal subscription form and a ballot for 2002 Executive Council positions. There is a deadline for the 2002 M&M Journal subscription of December 15, 2001 and the ballots must be postmarked by December 31, 2001. Everything can be returned in the envelope provided. Please take a few minutes to fill out the forms and ballot and drop them in the mail as soon as possible. This will help to avoid any future problems.
If you have not received your packet in the next few days, have any questions, or if you are not a member of the Microbeam Analysis Society and would like to join, please contact me at the email addresses or phone numbers below.
Thank you for your co-operation. Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.bioterc.missouri.edu/emc
Hello everybody Can you recommend any references that might have total emitted electron coefficient for different materials as a function of beam energy? I am interested in oxides, particularly in materials like PZT and BaTiO3. Thanks in advance Sergei
-- Sergei V. Kalinin Dept. Mat. Sci. Eng. University of Pennsylvania, 3231 Walnut St, Philadelphia PA 19104 Tel: (215) 898-3446 Fax: (215) 573-2128 E-mail: sergei2-at-seas.upenn.edu URL: http://www.seas.upenn.edu/~sergei2
A colleage of mine is inquiring if anyone knows of a person who can do routine servicing of MT2-B Porter Bloom microtomes in Central States area (central Illinois, St. Louis, Mo).
The only name they have is Bill McGee from New York. They are worried that getting someone from NY area would be very costly.
Respond directly to Dr. Chang at: Chang.Nada-at-uis.edu
OR, I will forward any responses.
Thank you.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
I made a drastic mistake in my earlier email announcement. Ballots must be postmarked by DECEMBER 1, 2001 to be counted. I apologize for the wrong date. All the more incentive to return your application, subscription and ballot as soon as possible.
Thanks, Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) www.microanalysis.org
Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.biotech.missouri.edu/emc
Food Structure & Functionality Symposium 2002 May 5 to 8, 2002, Palais des Congres de Montreal, Montreal, Quebec, Canada.
An international symposium leading Food Structure & Functionality studies through the 21st century "webaddress at the AOCS site (http://www.aocs.org/member/division/fsff/index.htm)" Held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
The mandate of the Food Structure & Functionality Forum : "To promote global collaboration between Food and Agriculture professionals in Structure and Functionality disciplines by facilitating and providing a forum for exchange of knowledge, expertise and research findings".
The symposium has two themes: * new and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Program Schedule Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl)
Monday, May 6th Opening of symposium Plenary Speaker Dairy Applications Session. Chairs: Mark Auty (mauty-at-moorepark.teagasc.ie) and Harjinder Singh (H.Singh-at-massey.ac.nz) Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl) and David Pechak (Dpechak-at-kraft.com)
Dedicated Poster Session Division Board Meeting
Tuesday, May 7th Agricultural Applications of Microscopy and Imaging Session./ joint with Feed Microscopy Division. Topic/Tentative title: New Microscopic Techniques for Identifying Food/Feed Constituents and Contaminants. contacts: Mark Auty (mauty-at-moorepark.teagasc.ie) and Kim Koch Division Luncheon and round table (expert) discussion. Topic TBA Microbiology and Food Session. Chairs Judy Arnold (jarnold-at-saa.ars.usda.gov) and Ida Yates (iyates-at-ars.usda.gov)
Division Members Meeting
Wednesday, May 8th Ingredients and Food Processing Session. Chairs: Diana Kittleson (dkittleson-at-pillsbury.com) and Bernhard Tauscher (bernhard.tauscher-at-bfe.uni-karlsruhe.de)
New Methods and Techniques for Food Structure and Functionality Analysis Session. Chairs: Kathy Groves (Kgroves-at-LFRA.co.uk) and Maud Langton (maud.langton-at-sik.se)
Closure of Symposium.
¯------------------------------------------------------------------------------------------------------------------ For more information visit the websites mentioned above, contact any of the session chairs listed above or myself.
Paula Allan Wojtas, Chair, Food Structure and Functionality Forum - a Division of AOCS
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Hi, We are trying to locate Mark Rigler of Materials Analytical Services. Have they gone out of business? Their 800# says "all circuits are busy now"...it has been like that for days. thanks for your help, Beth
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
Hi everybody, I am the operator of the SEM for my university and I am trying to shop for these two accesorries 1. Coater 2. EDX accesories The SEM is a LEO 1400. Soji
Hi All: I am looking for suggestions from experienced microscopists for voltage and current settings for the final cleaning to remove ion milling damage on Silicon and GaAs samples for High Resolution microscopy using a Gatan 600 ion mill. Presently we use the standard 5Kv, 0.5 milliamps/gun with "splotchy" and/or inconsistent results ( we are trying "tweak" our settings in order to reduce/eliminate the amorphous regions). Thanks in advance, Mike UT Arlington
The lowest you can go on the Duomill is about 10 degrees on the top side (with no top plate) and 12 degrees on the bottom side. With normal guns, you can go to about 2.5 - 3 kV with 2.5 mA.
If you insist on ion milling and you can only use the duomill, you can buy the PIPS sample holders and a pedestal from Gatan that can be used in the duomill to go to low angles. The lowest angles on dimple samples will be about 4 degrees on the dimpled side (assuming single sided dimpling). This should help you quite a bit. You will not have a lot of beam current to work with, but it should give you what you want with a little experimentation.
Now if you want no amorphous areas at all, (and I do mean none) you should use the small angle cleavage technique. Check out the Microcleave(TM) Kit from South Bay Technology on their web site. They have a couple of my PDF files that illustrate the quality of the samples that you can get.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Friday, October 05, 2001 3:47 AM To: listserver
Hi All: I am looking for suggestions from experienced microscopists for voltage and current settings for the final cleaning to remove ion milling damage on Silicon and GaAs samples for High Resolution microscopy using a Gatan 600 ion mill. Presently we use the standard 5Kv, 0.5 milliamps/gun with "splotchy" and/or inconsistent results ( we are trying "tweak" our settings in order to reduce/eliminate the amorphous regions). Thanks in advance, Mike UT Arlington
I look after a JEOL 840 with EDS and 3 x WDS in a small department.
I usually put it in standby and there's no useage while I'm away, but I'm being urged to train a deputy to mind it and help/supervise users.
But when I think about the possibility of returning to find the column and/or the spectros full of oil, or some other nightmare scenario that I (thankfully) haven't yet envisaged, I become apprehensive.
How do other places/people handle the problem of absences of the primary/sole caregiver?
thanks
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Does anyone have any information on Schiff's reagent that contains thionine or any other blue stain. If so who is the manufacturer, who is the distributor, what's the cost, what's the shelf life, etc., etc.. I am looking for an alternative Feulgen stain for DNA ploidy analysis via Image Analysis.
Thank you in advance.
Donald G. Awbrey, HT(ASCP) QIHC Image Analysis / Electron Microscopy donaldawbrey-at-texashealth.org donaldawbrey-at-hotmail.com
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, October 06, 2001 2:47 AM
Dear Listers,
I have a JEOL 35C with a defective high voltage cable. It appears to be "open" at the feedthrough to the gun. As the SEM is quite old JEOL no longer supplies replacement cables.
Morning Don, But the Feulgen Schiff has been proved to be stoichiometric! There are alternatives, but you would have to prepare them yourself and insure that the stoichiometry holds, but, then, I have always made my own Schiff reagent anyway. (Method on request, but it is readily available.) [A.G.E. Pearse, Histochemistry, Theoretical and Applied, (4th Ed.), Churchill Livingstone, New York, 1985, Vol II, ISBN: 0-443-02997-0] always has plentiful methods, and the most recent edition (Vol II) not only has many methods [including alternatives to Schiff!] but a DEEP discussion of the entire Feulgen-DNA methodology and history. Hope this helps.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Awbrey, Donald } Sent: Friday, October 5, 2001 6:41 PM } To: 'microscopy-at-msa.microscopy.com' } Subject: Schiff Reagent for Image Analysis } } ------------------------------------------------------------------------ } 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 histo/micro netters, } } Does anyone have any information on Schiff's reagent that contains } thionine } or any other blue stain. If so who is the manufacturer, who is the } distributor, what's the cost, what's the shelf life, etc., etc.. I am } looking for an alternative Feulgen stain for DNA ploidy analysis via Image } Analysis. } } Thank you in advance. } } } } Donald G. Awbrey, HT(ASCP) QIHC } Image Analysis / Electron Microscopy } donaldawbrey-at-texashealth.org } donaldawbrey-at-hotmail.com } } }
We recently purchased a DVD RAM drive to assist with image storage. The problem is that our IT department and local venders don't know much about the media (The actual disks). We have two choices for DVD RAM disks: authoring or general use. Has anyone out there figured out which is the best for general microscopy images. The disks will travel between Mac's, PC's and LINUX/UNIX operating systems.
TIA,
Jon Ekman Florida State University Biological Science Imaging Resource 119 Bio Unit I, 4370 Tallahassee, FL 32306 tel: 850.644.6519 fax: 850.644.0481
John H. L. Watson Memorial Student Scholarships 15th International Congress on Electron Microscopy (ICEM-15), Durban, South Africa, Sep 1-6, 2002
In memory of John H. L. Watson, the Microscopy Society of America (MSA) has established a Travel Scholarship Award to encourage graduate students and post-doctoral research assistants who are MSA members in good standing, to attend international microscopy meetings. (Note: In the future, The John L. Watson Memorial Student Scholarships should also be available to enable young researchers from Latin America and South America to attend appropriate international meetings hosted by the MSA in the USA).
The MSA will pay from $500 to $1250 towards travel and accommodation to allow graduate students or post-doctoral research assistants (with Doctoral degrees awarded on or after January 1, 1998) who are members of the MSA to attend the 15th International Congress on Electron Microscopy (ICEM-15), in Durban, South Africa, September 1-6, 2002. The level of the monetary award will be based on the scientific content of the paper to be presented and on the number of successful applicants.
Applicants should forward to the MSA Awards Committee Chair, Dr. J. Bentley, MS-6136 4500-S, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Oak Ridge, TN 37831-6136: (i) their full name, age, address and details of academic and/or professional status, (ii) a letter of support from their advisor/supervisor that also attests to the eligibility of the applicant, and (iii) an "advanced" copy of their 2-page paper/extended abstract intended for submission to ICEM-15, on which the applicant is the lead author.
The deadline for submission is December 15, 2001. This date will allow time for evaluation of the paper by the MSA Awards Committee, confirmation with the MSA Business Office that the applicant is a member in good standing and has renewed their dues for 2002, selection/approval by MSA President, and notification of successful and unsuccessful applicants in time to meet the submission deadline of papers to ICEM-15 (February 1, 2002). Successful applicants will be required to submit an appropriate paper/extended abstract in sufficient time and of sufficient scientific content to be published in the ICEM-15 proceedings. Unsuccessful applicants will be encouraged to submit an abstract to ICEM-15, but will receive no financial support from this award.
__________________________________________________________________________ Jim Bentley Microscopy Microanalysis Microstructures Group Metals and Ceramics Division Bldg. 4500-S, MS-6136 Oak Ridge National Laboratory PO Box 2008 Oak Ridge, TN 37831-6136, USA
Tel: (865)574-5067 Fax: (865)241-3650 bentleyj-at-ornl.gov express mail use "1 Bethel Valley Rd" instead of PO Box. ** Note the new group name, mail-stop, fax, and area code **
General use media is all that you need. There are two types, Type 1 5.2GB and Type 2 2.6GB. Type 1 is double sided and can only be used in its container. Type 2 can be used in the container or removed for reading in DVD-ROM drives.
Check your drive and media carefully. I've used Panasonic and Pioneer drives, TDK and Maxell media. When writing data to the media, I had no problem. When reading the data from the media, many files were corrupted. Any file larger than about 1MB was likely to be damaged. Be sure to perform a verify after writing. Routine scandisk is also a good idea.
I don't use DVD-RAM any more since I cannot trust them. I switched to DVD-R and DVD-RW. No problems with this product.
gary g.
At 08:50 AM 10/6/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm particularly interested in an electron microscopy application, hope someone out there can help or point me in the right direction.
What I want to do is follow the route of a phagocytosed bacterium from the cell surface attaching initially to the cell surface or pseudopods to being engulfed right through to it's final destination within a lysosome and all the intervening steps in between. I have a very good antibody to a cell surface antigen on the bacteria which has been shown to work well in an immunogold labelling previously so I'd like to try and repeat this.
As I have no experience with TEM or indeed preparation of the samples I would appreciate very much any useful comments or protocols on this matter. I intend to grow the cells in flasks then add the bacteria and take a variety of time points to catch the range of stages of the bacteria through the cell - is there perhaps a better way to setup this experiment ? How might I collect the sampes, ie cell scraping or limited ezymatic digestion to get the cells of the cell culture plat ? I really want to preserve cell morphology as much as possible and the show the very early contacts between macrophage & bacterium at the cell surface so am concerned about the best way to harvest the cells, ie enzymatic harvseting or cell scraping may be worrysome. Is it possibe to process the cells for TEM & immunogold analysis whilst still attached to the plastic of a cell culture flask (ie break the flask open and somehow embed/ process the cells whilst still in-situ ?).
Any useful comments would be greatly appreciated !
In Europe when I was hopping from country to country I found that the organisations that work with X-ray sets in hospitals etc are more than capable of fixing a high voltage cable. We have even replaced the complete cable simply taking the two ends from the EM cable.
Hope this helps?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
You bring up a very good issue of removing amorphous damage from specimens which has become of increasing interest in HREM applications. Removing amorphous damage from specimens has been most recently addressed with the advent of low energy ion milling. This is a technique where the final milling stage is done at ~ 100-500eV. This is based on technology developed by Dr. Arpad Barna in Budapest and has shown some tremendous results. We market the aptly named "Gentle Mill" and you can find results for both silicon and GaAs on our website at www.southbaytech.com. Go to the search function and enter "gentle" and that will bring you to the right page. You need to register to download the PDF - if that's a bother, just send me an email and I will email the file to you.
I am sorry for the somewhat commercial nature of the response, but I do think it appropriately addresses the issue. I hope it helps.
Best regards-
David
"Walck, Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The lowest you can go on the Duomill is about 10 degrees on the top side (with no top plate) and 12 degrees on the bottom side. With normal guns, you can go to about 2.5 - 3 kV with 2.5 mA. } } If you insist on ion milling and you can only use the duomill, you can buy the PIPS sample holders and a pedestal from Gatan that can be used in the duomill to go to low angles. The lowest angles on dimple samples will be about 4 degrees on the dimpled side (assuming single sided dimpling). This should help you quite a bit. You will not have a lot of beam current to work with, but it should give you what you want with a little experimentation. } } Now if you want no amorphous areas at all, (and I do mean none) you should use the small angle cleavage technique. Check out the Microcleave(TM) Kit from South Bay Technology on their web site. They have a couple of my PDF files that illustrate the quality of the samples that you can get. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } P. O. Box 11472 (letters) } Guys Run Rd. (packages) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8515 (fax) } } -----Original Message----- } } From: coviello [mailto:coviello-at-mae.uta.edu] } Sent: Friday, October 05, 2001 3:47 AM } To: listserver } Subject: TEM-HRTEM samples with Gatan 600 ion mill } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi All: } I am looking for suggestions from experienced microscopists for voltage } and current settings for the final cleaning to remove ion milling damage } on Silicon and GaAs samples for High Resolution microscopy using a Gatan } 600 ion mill. Presently we use the standard 5Kv, 0.5 milliamps/gun with } "splotchy" and/or inconsistent results ( we are trying "tweak" our } settings in order to reduce/eliminate the amorphous regions). } Thanks in advance, } Mike } UT Arlington
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
We are using magneto-optical disks (MO-disk) for 5+ years. Since that time we had 1 disk damaged (and successfully recovered) out of few hundreds which is in circulation currently. Each disk is 2.6 or 5.2 Gb and 9-something available now. The disk uses FAT or NTFS format. DVD format is not established well and a subject of frequent changes over last few years, so I would avoid it unless they establish standard and will follow it. "FAT"- disks are readable from any platform and no software necessary to operate the drive (it's SCSII) - it's visible on desktop immediately after connection.
Sergey
At 01:29 PM 10/6/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Dear All, We recently purchased a new Leica EM Stainer to replace our aging (but otherwise pretty trusty) LKB Ultrostainer and have some misgivings about our new acquisition. Until the company has had a bit longer to try to sort the problems I do not want to go into specifics here but despite of spending several months trying to sort out theses 'issues' we are a long way from being happy with the instrument - this is despite it having been back to the factory once.
I would really like to know if other owners and users of the new EM Stain have experienced major problems with this piece of equipment - or if its just our one (or us!). If your EM Stain has proved totally satisfactory, I'd like to hear from you too...
Best regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
Nextek Inc. is seeking a qualified individual to join our analytical laboratory team as a Materials/Failure Analysis Engineer. The ideal candidate will have a B.S. in Materials Science, Chemistry, Engineering or equivalent, and a minimum 3 years experience in the micro-electronic field; or 5 or more years hands-on lab experience in the micro-electronic materials/failure analysis field. Knowledge in cross-sectional and destructive analysis of micro-electronic assemblies and devices, and experience with analytical instrumentation such as; SEM/EDS, Micro-FTIR, Thermal Analysis, Acoustic and Optical Microscopy is a plus.
To learn more about Nextek Inc. visit our web site at http://www.nextekinc.com/. For more information about this position, call or email the following:
Wade McFaddin (256) 772-1995 ext.1064 wade.mcfaddin-at-nextekinc.com or Jim Chiang (256) 772-1995 ext. 1029 james.chiang-at-nextekinc.com
Dear All: I am looking for a monoclonal anti pancreatic amylase antibody. Does anyone out there know a source. Thank you in advance.
Hong ------------ Hong Yi Emory Neurology Microscopy Core Laboratory Emory University Department of Neurology 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
Many years ago, when (or even before) I was an undergraduate (that would place it in the early '60s), my aunt managed the analytical lab at a company that produced non-ferrous metals (principally, I think, bronzes and brasses). As one does in such situations, she invited me to tour her labs one day.
One of the instruments I was shown was called a "Beta Probe". A piece of the metal was ground and polished, and was then placed face down on an aperture in the top plate, with what I now realize was an O-ring seal. A few buttons were pressed, and after a little while some of the electronic counters of the day (neon tubes with 10 electrodes) began spinning round. My memory is that the operator had some range of ratios between the recorded numbers within which he (it was, of course, a "he" in those days, even though my aunt was the manager) could pass the sample as meeting specifications.
I surmise that this was an electron milli-probe (if you'll forgive my coining of the term!) with preset spectrometers tuned to the elements of interest in the specification. I would very much like to find out more about this device. Happily, my aunt is still available to be asked, but she knows nothing of the operation of the instrument. The people who ran it have either died, or lost touch (the company failed twenty years ago).
If anyone can give me more information about this instrument, I would be very interested, and most grateful.
The University of New Hampshire is looking for a Director of the Instrumentation Center. For more information please go to our website at:
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Analytical services at the UIC include: Elemental Analysis; Gas Chromatography-Mass Spectrometry (GC-MS); High-Field Nuclear Magnetic Resonance (FTNMR - 90, 360, 400, and 500 MHz); Ultra-Violet, Visible, and Near-Infrared Spectrophotometry (UV-Vis-NIR); Infrared Spectroscopy (FTIR); Optical Rotation; X-Ray Photoelectron Spectroscopy; Electron Microscopy including Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) with Energy Dispersive Spectroscopy (EDS).
RESPONSIBILITIES: Reporting to the Vice President for Research and Public Service, the University Instrumentation Center Director will be responsible for leading an emerging research and teaching oriented service center toward excellence; duties to include:
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Supporting education and training of staff, university researchers/clients, and students;
Securing funding through external and internal sources in order to obtain new instruments and upgrade existing equipment;
Promoting UNH outreach mission by developing knowledge and awareness of other laboratories in the state and region, for purposes of forming partnerships, referral relationships, information networks, etc. Serving as a resource for state government and business and industry.
QUALIFICATIONS:
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Minimum of three years managerial experience.
Demonstrated experience with operation of analytical instruments and application to research problems.
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STARTING DATE: Negotiable, as early as 12-1-01
SALARY: Competitive and commensurate with experience and local area.
APPLICATION DEADLINE: Open until filled.
APPLICATION PROCEDURE: Applicants should send a letter of intent, CV/resume, and the names, addresses, and phone numbers of three references to:
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University of New Hampshire
Durham, NH 03824
Electronic resumes and supporting documentation will be accepted. Please e-mail your package to: diana.couture-at-unh.edu
Can anyone offer any ideas on the best way to proceed with removing the effects of charging on an image? I am referring to the streak across the image manifested as a change in density. As an example, have a look at this uncoated mite:
http://katie.ucdavis.edu/pics/sucker2.jpg
The nature of this projects requires that we place the live mite in the scope and get a picture in the first 5 minutes. This picture was taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor for publication. I am looking for some help in eliminating some of the charging artifacts without losing too much detail. Any help will be appreciated.
We are using PS 6 and I have the IPTK plugins. I made some progress using a Gaussian filter to produce a very fuzzy duplicate image, invert the image, add it to a layer and then adjust the opacity to around 50%. This was only partially successful. Any help will be appreciated.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
I would be interested to hear of any IP solution you come up with.
However, I would think that running the scope in environmental mode at higher voltage ought to be able to give you a satisfactory image.
We have a Hitachi S2460N here. We routinely run 40 Pa of Helium to eliminate charging. At more than 10 kV, we can get a suitable image off of our Robinson BSE. Our Oxford Tetra(tm) allows us to drop the voltage further. I will forward you an image of a bug from our scope under separate cover.
Warren
At 09:29 AM 10/9/2001 -0700, you wrote:
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
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In a message dated 10/9/01 12:45:01 PM, raharris-at-ucdavis.edu writes:
} We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated.
Another approach you might try is to duplicate the image, smooth the copy with a Gaussian blur, and then divide by it. The size of the blur to use depends on the size of the bright streaks, and these are not consistent in your image, but values in the range of 2-4 pixels seem to work fairly well.
Hi Rick, If the image is saturated with white pixels ("blown out highlights") then your are pretty stuck. There isn't anything that can be done to put information in where it doesn't exist. However, in order to make the most out of a bad situation, you can use the levels and curves options to adjust the brightness and contrast for a reasonable image. I prefer to use adjustment layers to do this, that way I can edit and tweak the settings without permanently changing the core data. Your best best is to avoid/minimize saturation when you capture your images. A "wet" ESEM is fantastic for this. As you probably know, try to minimize spot size, current, and voltage to minimize charging artifacts, and choose brightness and contrast settings that don't saturate the image. If you want to see what I mean by adjustment layers using curves and levels, you can send me a file, and I'll set it up for you, and send it back for you to look at. Good luck.
-Brad
---------- From: Rick Harris Sent: Tuesday, October 9, 2001 9:29 AM To: Microscopy-at-sparc5.microscopy.com Subject: Photoshop and SEM charging
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Can anyone offer any ideas on the best way to proceed with removing the effects of charging on an image? I am referring to the streak across the image manifested as a change in density. As an example, have a look at this uncoated mite:
http://katie.ucdavis.edu/pics/sucker2.jpg
The nature of this projects requires that we place the live mite in the scope and get a picture in the first 5 minutes. This picture was taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor
for publication. I am looking for some help in eliminating some of the charging artifacts without losing too much detail. Any help will be appreciated.
We are using PS 6 and I have the IPTK plugins. I made some progress using a Gaussian filter to produce a very fuzzy duplicate image, invert the image, add it to a layer and then adjust the opacity to around 50%. This was only partially successful. Any help will be appreciated.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Rick, My experience is that live insects don't generally charge because the evaporating water carries the charge away. From the looks of the image, I think you were using a lot of beam current. Cur that back a whole lot and the charging will probably go away. Also, the horizontal scan speed has a much greater effect on charging than the vertical speed. If you run a faster horizontal rate and more lines (slower vertical speed), this will help.
Ken Converse owner Quality Images third party SEM service Delta, PA
Rick Harris wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone offer any ideas on the best way to proceed with removing } the effects of charging on an image? I am referring to the streak } across the image manifested as a change in density. As an example, } have a look at this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in } the scope and get a picture in the first 5 minutes. This picture was } taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but } is too poor for publication. I am looking for some help in } eliminating some of the charging artifacts without losing too much } detail. Any help will be appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress } using a Gaussian filter to produce a very fuzzy duplicate image, } invert the image, add it to a layer and then adjust the opacity to } around 50%. This was only partially successful. Any help will be } appreciated. } } } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu } } } }
Our facility is looking to purchase a dry 1x objective for a Nikon Microphot light microscope. I have contacted a few vendors and have been told that the microscope is out of date and it will be impossible to locate such an objective. Any help in locating this item will be appreciated.
Twin City International Inc. made a beta backscatter instrument called the Betascope. It used an isotope source and a Geiger-Mueller tube to measure the thickness of metal films after a simple calibration with known thickness standards. I don't know if they are still in business.
-----Original Message----- } From: Tony Garratt-Reed [mailto:tonygr-at-mit.edu] Sent: Tuesday, October 09, 2001 7:59 AM To: microscopy-at-sparc5.microscopy.com
Hi, Listers!
Many years ago, when (or even before) I was an undergraduate (that would place it in the early '60s), my aunt managed the analytical lab at a company that produced non-ferrous metals (principally, I think, bronzes and brasses). As one does in such situations, she invited me to tour her labs one day.
One of the instruments I was shown was called a "Beta Probe". A piece of the metal was ground and polished, and was then placed face down on an aperture in the top plate, with what I now realize was an O-ring seal. A few buttons were pressed, and after a little while some of the electronic counters of the day (neon tubes with 10 electrodes) began spinning round. My memory is that the operator had some range of ratios between the recorded numbers within which he (it was, of course, a "he" in those days, even though my aunt was the manager) could pass the sample as meeting specifications.
I surmise that this was an electron milli-probe (if you'll forgive my coining of the term!) with preset spectrometers tuned to the elements of interest in the specification. I would very much like to find out more about this device. Happily, my aunt is still available to be asked, but she knows nothing of the operation of the instrument. The people who ran it have either died, or lost touch (the company failed twenty years ago).
If anyone can give me more information about this instrument, I would be very interested, and most grateful.
Does anyone know of a dye that binds to cellulose (or pectins) and is fluorescent? I am looking for an alternative to calcofluor. I wish to stain 1 micron thick LR-white sections of Vicia faba cotyledons.
Hi there, I know this question has been asked lots, but i never needed to pay any attention till now, so my apologies. My institute wishes to purchase a very good digital camera for light / fluorescence microscopy to replace (compliment?) the 35 mm film cameras that we currently use. Part of the requirements is that the camera be able to be taken off the scope and have a regular lens fitted for other high / professional quality images (such as whole plants, animals etc.) So the question I have to ask is, is this a reasonable task for a system? Further to that, what do current users of digital cameras for microscopy see as an absolute requirement and what is nice to have?
A short list of cameras suggested to us locally are: Canon D30 3.3 megapixel, Fuji/Nikon S1 3.3mp, Nikon DiX 5.5 mp. Any comments on those (good or bad)? Any other recommendations?
John Blinco Western Australian State Agricultural Biotechnology Center
Mark Congo red would be a suitable alternative. Excite with green and view in orange/red - Rhodamine -type filter set.
You could also try a fluorescent pseudo-schiff reaction using periodic acid or sodium m-periodate oxidation followed by staining with a carbohydrazide derivative of a fluorescent dye - e.g. Lucifer Yellow CH.
A dye suitable for uronic acid polysaccharides such as pectin is Coriphosphine O CI No. 46020. Again this works with Rhodamine type filter sets
Chris
Date sent: Wed, 10 Oct 2001 12:21:11 +1000 } From: MARK JEFFREY TALBOT {mark.talbot-at-studentmail.newcastle.edu.au}
The Hitachi S3500N has the N-SEM mode which should give you a reasonable image in backscatter, with appropriate WD and pressure settings. Also the 3500 has an integrate capture mode to do multiple rapid scans which can help reduce charging.
David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/micros_e.html
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: October 9, 2001 1:30 PM To: Microscopy-at-sparc5.microscopy.com
Can anyone offer any ideas on the best way to proceed with removing the effects of charging on an image? I am referring to the streak across the image manifested as a change in density. As an example, have a look at this uncoated mite:
http://katie.ucdavis.edu/pics/sucker2.jpg
The nature of this projects requires that we place the live mite in the scope and get a picture in the first 5 minutes. This picture was taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor for publication. I am looking for some help in eliminating some of the charging artifacts without losing too much detail. Any help will be appreciated.
We are using PS 6 and I have the IPTK plugins. I made some progress using a Gaussian filter to produce a very fuzzy duplicate image, invert the image, add it to a layer and then adjust the opacity to around 50%. This was only partially successful. Any help will be appreciated.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
One of those answering Rick Harris' question on charging remarked about using helium in the variable pressure mode. What other gasses are used for the VPSEM and ESEM? What considerations are taken into acccount in choosing a vent gas? Is there a paper somewhere that answers these questions?
We use Congo Red and image it in the rhodamine channel...
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One of our pharmacology researchers is making nanoparticles ranging in sizes from 50nm up to 250nm. The material is polylactic-acid-coglycolic-acid (PLGA). He's looking for a lipophilic electron dense particle, dye or stain that could be incorporated into the particles during their formation and which could be seen in the TEM sections. The material has to be able to be suspended or dissolved in an organic solvent.
For our part here are some of the things we've thought of using: Ruthenium Red powder, colloidal gold in 5 to 10nm range, phosphotungstic acid in ethanol.
His material will also melt at 45 degrees centigrade. So I embedded in Unicryl, UV polymerization at 4 degrees centigrade. Does anyone know if there is still heat produced during the polymerization?
This seems to be in the realm of polymer chemistry and materials science. I'd appreciate any and all help, Thanks.
We run a Hitachi S2250N. Rick, what gas, pressure and WD were you using? Helium is the best, as Warren says, but if you dont have a tank handy, water vapour (for a minimal system - just connect the gas inlet to a half-full flask of water!) is almost as good for imaging. And I would also suggest a higher voltage - 5kV minumum, 10-15 for comfort, if you want to work quickly at low magnifications and can afford to miss a bit of surface detail. On something with as many projections as a mite, utilising the VP system to eliminate charging will let you work much more rapidly than relying on low kV alone. Its not very PC, but we would tend to start at 20kV and work down....
regards Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
} } } Warren E Straszheim {wesaia-at-iastate.edu} 10/10/01 03:30AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would be interested to hear of any IP solution you come up with.
However, I would think that running the scope in environmental mode at higher voltage ought to be able to give you a satisfactory image.
We have a Hitachi S2460N here. We routinely run 40 Pa of Helium to eliminate charging. At more than 10 kV, we can get a suitable image off of our Robinson BSE. Our Oxford Tetra(tm) allows us to drop the voltage further. I will forward you an image of a bug from our scope under separate cover.
Warren
At 09:29 AM 10/9/2001 -0700, you wrote:
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I'm not an expert at this, but I do play with images in photoshop for my own amusement.
I played with your image in Photoshop and the best I could do was reduce the difference in darkness between the top and the bottom of the picture by lasooing the top portion of the picture (at 200X mag.) with the lasoo tool and then using the brightness-contrast setting under Image, Adjust, to get the top and bottom portions of the photo to match in brightness. This still left the lines, which I tried the gausian blurr on, but it couldn't be blurred much before it looked out of place. (like a blurry band in the middle of an otherwise crisp the picture.)
This is just another trick you might try. Good luck.
Karen Pawlowski
Rick Harris wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu
} Dear Listers, } } Thank you to all who have responded to my request for high voltage cable repair. } } The results were: } } 1. Fix it yourself. } 2. Buy another JEOL 35C for the cable. } 3. Have a third party repair the cable. } } The consensus is that the best third party repair is: } } } Dielectric Sciences Inc. } } 88 Turnpike Road } } Chelmsford, MA 01824 } } Phone: 978 250-1507, FAX: 978 250-1699 } } } Best Regards, } } Earl Weltmer
We need some help for solving a problem related to the deformation of the image taken with our SEM . We have already asked a number of people about this but, it seems the solution is not so straightforward.
We have a JEOL SEM 35C, from the early 804s. Recently, we installed the SemAfore digital acquisition system which gets the electronic signal directly out of the column and sends it to the computer. The CRT4s show the same analogic image as always.
The distorsion of the image appears at the beginning of every scanline ramp (lefthand side on the image). Two micrographs digitally taken from latex microspheres give the idea what the problem looks like: the same feature (a group of microspheres) is taken positioning them at the very left-hand side of the image, then moved to the right-hand side of the same CRT screen. It is clear that the distortion on the shape of the spheres appears on the first image (on the second one, you see a very very little deformation to the opposite direction). Find both micrographs at:
http://www.ceride.gov.ar/servicios/sem/kuqui1.jpg
http://www.ceride.gov.ar/servicios/sem/kuqui2.jpg
The technicians at our institute tested with the oscilloscope the shape of the signal out of the scan generator unit. See, at:
http://www.ceride.gov.ar/servicios/sem/ the file: dibujo_mail.bmp
a drawing is seen: in blue the theoreticall scan generator ramp, while in magenta the real one as shown on the oscilloscope. The scanning ramp showes a distortion on the shape starting from the begining up to the 8% of the leght of the ramp, along a scanline (see the magenta dip inside the green square). From that point, the ramp followes the shape that should have up to the end of the scanline. Rounded corners at the beginning and the end of the ramp are commonly found, according to the technicians.
This problem was seen on the CRT screen since long ago, but since the image on the CRT is diminished in size, the defect is not so obvious nor so serious. The digital imaging system gets and showes on the PC screen the complete image, made up by the complete scan along a line and, along the frame. In these conditions, the distortion shown on the digital images is dramatical.
The technicians say that a sort of over scan made via software, once the image is collected, could help. But, that solution reduces the side of the viewing field and takes extra imaging processing work.
Any suggestion or help will be very much appreciated, thanks in advance.
Silvia Montoro- Nora Pratta Centro Regional de Investigaciones y Desarrollo de Santa Fe (CERIDE) G|emes 3450 3000 Santa Fe Argentina csedax-at-ceride.gov.ar ---------------------------------------------------------------------------- ----
I have unearthed 2 EM film casettes (the boxes that store film in a TEM). They have no identifying marks. They are black, about 5 1/2 x 4 3/8 x 2 1/4 inches. The black metal top that slides out is about 5 1/2 x } 6 7/8 inches. They do not belong to Philips or JEOL microscopes, but I cannot determine where they originated, perhaps Zeiss? or Hitachi? If anyone thinks (s)he can use them, please reply directly, and I will send them to you. Otherwise, they will have to take up some space in the landfill.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Your effect looks like there is not enough fly-back delay for your SEM under your present operating conditions. We can see the same foreshortening and shifting on the monitor of our JEOL 840A as we switch through the various scan speeds. The effect is particularly pronounced at TV-rate scans. It is present, but not so bad, at the SR (super-rapid?) scan rate, and it is not apparent at the Slow-1 or Slow-2 scan settings.
I am not familiar with the SemAfore system. Is is an active system (takes control of the SEM scan) or a passive system (just digitizes the regular SEM scan)? If it is an active system, there should be an adjustment to increase the fly-back delay before the next horizontal scan starts. If they don't provide such an adjustment, they should add one. If it is a passive system, then you should slow down your SEM's scan.
Warren
At 12:27 PM 10/10/2001 -0300, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Dear colleagues, I don't know if this thread has already come up, but here goes: Does anyone have tips on using EM to identify transplanted cultured cells labelled with latex beads in a host animal. Especially to see if synapses are being formed. Would traditional methods destroy the latex? if so, are there other labels for the cultured cells? Thanks in advance, Julie Gross Dept. of Neuroscience UCONN Health Center Farmington, CT 06030
1. Brad Johnson wrote: "Your best bet is to avoid/minimize saturation when you capture your images A "wet" ESEM is fantastic for this. As you probably know, try to minimize spot size, current, and voltage to minimize charging artifacts..."
Your S3500N is well equiped to do this. Consistent with the above suggestions, have you also been playing around with the mites in the variable pressure mode? In VP mode, use your backscattered electron image which inherently shows much less charging artefact thae SE imaging, and by adjusting the pressure you may be able to eliminate charging. In BSE mode, you may need higher kV, like 5-10 kV, to "see" through the gas in the VP mode, and in order for BSE's to be detected by your BSE detector.
If you have the ESED mode on your S3500N, that will enable you to to get a SE-like image in VP mode. Still, at the low mags you are working at, BSE should do fine for sufficent resolution, and may be easier to get rid of charging in that mode.
You may even be able to succeed using BSE image in high vacuum mode, if the mite doesn't dry out too fast..
2. Ken Converse wrote: "Also, the horizontal scan speed has a much greater effect on charging than the vertical speed. If you run a faster horizontal rate and more lines (slower vertical speed), this will help."
Again on your S3500N, you have that Image Integration mode that often can be used to eliminate charging in whatever scope mode you are operating in. Select the slowest line scan rate such that you observe no charging bands on your monitor. If you still see some bands, select next fastest rate. Then go to Image Integration set-up and select the number of frames to add tgether, each recorded at the scan rate you just set, usually something like 4-10 will work.
Hope this helps, and good luck. Let us all know what works for you.
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image?...snip!...
} The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated.
} } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu
} This problem was seen on the CRT screen since long ago, but since the image } on the CRT is diminished in size, the defect is not } so obvious nor so serious. The digital imaging system gets and showes on the } PC screen the complete image, made up by the complete scan along a line and, } along the frame. In these conditions, the distortion shown on the digital } images is dramatical.
I think your clue is in the statement above. If the defect can be seen on the SEM's CRT screen, no mater how small, then your SEM has a scan generator problem. SemaAfore (a JEOL product) is a passive scan digitizer. It can only do as good as the 35C's scan generator. Your 35C's scan generator is producing ramps that are non-linear at the start and end of the ramp. Like the tech said, thats not abnormal.
What's missing from your ramp diagram is where does the SEM starts using the ramp signal for image display. What happens is the ramp is non-linear at the start but the SEM has a little delay after the start to get into the linear range before using the ramp for image display. The horizontal blanking signal defines this region.
You need to look at the horizontal blanking signal to see the real start/end of ramp location. If you are still non-linear in that ramp region, then the tech needs to fix this first on the SEM.
SemAfore might have a software param to add a delay after the horizontal blanking signal. This might fix the acquired image but you are better off fixing the SEM first.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (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
Dear list member, I am looking for an old paper, Th. Koller, et al., Micron 1:110, (1969). This journal is not available in Hopkins. Does anyone would lile to help me? thanks
Chen Chen
Department of Biological Chemistry The Johns Hopkins University School of Medicine 725 N. Wolfe Street Baltimore, Maryland 21218
Rick, I am assuming you want to fix this image rather than go collect another one. It depends on how much time and effort you wish to put into this project. One way, that I have used rather successfully has been a combination of the clone tool with a small brush size and just a little feathering of the edge, brightess/contrast adjustment, and a little spot blurring. Do each change as a separate layer leaving your original intact. It will take some time but I can show you before and after images (much worse than yours since molds tend to charge like heck) that you'd never know where charging.
Dr.Tina S. Schwach, President Microscopy Consulting Services Inc. 651-681-0112
Hello Silvia, We recently had an EDS mapping and imaging upgrade done to our SEM. When the technician installed the imaging hardware and software, there were several adjustments made (using software settings) to select the field of view that was digitally captured. As I watched the field engineer work, it was apparent that the area selected for image capture was smaller than the scanned area. Roughly speaking, it appeared that the scanned area was about 10-15% larger than what was captured. Possibly you could make some adjustments to the image capture area such that your digital image field of view would be the same as your analog, while retaining a larger scanned area. Since every manufacturer has their own way to accomplish these things, you would probably need to get the specifics directly from them. Good luck.
-Brad
---------- From: Nora Pratta- Silvia Montoro Sent: Wednesday, October 10, 2001 8:27 AM To: microscopy-at-sparc5.microscopy.com Cc: csedax-at-ceride.gov.ar Subject: SEM: distortion on digital images
------------------------------------------------------------------------ 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 members of the list,
We need some help for solving a problem related to the deformation of the image taken with our SEM . We have already asked a number of people about this but, it seems the solution is not so straightforward.
We have a JEOL SEM 35C, from the early 804s. Recently, we installed the SemAfore digital acquisition system which gets the electronic signal directly out of the column and sends it to the computer. The CRT4s show the same analogic image as always.
The distorsion of the image appears at the beginning of every scanline ramp (lefthand side on the image). Two micrographs digitally taken from latex microspheres give the idea what the problem looks like: the same feature (a group of microspheres) is taken positioning them at the very left-hand side of the image, then moved to the right-hand side of the same CRT screen. It is clear that the distortion on the shape of the spheres appears on the first image (on the second one, you see a very very little deformation to the opposite direction). Find both micrographs at:
http://www.ceride.gov.ar/servicios/sem/kuqui1.jpg
http://www.ceride.gov.ar/servicios/sem/kuqui2.jpg
The technicians at our institute tested with the oscilloscope the shape of the signal out of the scan generator unit. See, at:
http://www.ceride.gov.ar/servicios/sem/ the file: dibujo_mail.bmp
a drawing is seen: in blue the theoreticall scan generator ramp, while in magenta the real one as shown on the oscilloscope. The scanning ramp showes a distortion on the shape starting from the begining up to the 8% of the leght of the ramp, along a scanline (see the magenta dip inside the green square). From that point, the ramp followes the shape that should have up to the end of the scanline. Rounded corners at the beginning and the end of the ramp are commonly found, according to the technicians.
This problem was seen on the CRT screen since long ago, but since the image on the CRT is diminished in size, the defect is not so obvious nor so serious. The digital imaging system gets and showes on the PC screen the complete image, made up by the complete scan along a line and, along the frame. In these conditions, the distortion shown on the digital images is dramatical.
The technicians say that a sort of over scan made via software, once the image is collected, could help. But, that solution reduces the side of the viewing field and takes extra imaging processing work.
Any suggestion or help will be very much appreciated, thanks in advance.
Silvia Montoro- Nora Pratta Centro Regional de Investigaciones y Desarrollo de Santa Fe (CERIDE) G|emes 3450 3000 Santa Fe Argentina csedax-at-ceride.gov.ar
Dear List, It seems the meter, model 505, for my Edwards penning gauge has bit the dust. Edwards no longer sells nor works on the 505's so getting it repaired doesn't seem to be an option. Would anybody perhaps have one of these "obsolete" meters gathering dust on their shelf? I would appreciate hearing from you. Thanks. Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
I just heard about Two-Photon (or Multi-Photon) excitation microscopy. Does anybody know of a web site that has basic information about this technique, descriptions of applications, and the advantages. Something like "Two-Photon Microscopy for Dummies" would do.
This has always been such a terrific source of information.
Has anyone used the anti-SKI antibody from Santa Cruz? Or any other commercial antibodies to SKI (and/or SKIP)?
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
The distortion is caused by the scan coils "ringing".
When an SEM (or imaging system) acquires an image, it rasters from left to right then restarts at the left side of the screen again.
It is the "retrace", from right to left, at a relatively high speed that causes the scan coils to generate "back emf" or "ringing".
SEM manufacturers usually "blank" ,or turn off, the video during the retrace and part of the first few milliseconds of video. The effect is that you lose part of the image that is distorted.
What can be done to resolve your problem? Slower horizontal scan speed would help. Better yet, ask the vendor to blank the image just as the SEM manufacturers do.
Hope this helps,
Earl
----- Original Message ----- } From: "Johnson, Bradley R" {Bradley.Johnson-at-pnl.gov} To: {microscopy-at-sparc5.microscopy.com} ; "'Nora Pratta- Silvia Montoro'" {csedax-at-ceride.gov.ar} Sent: Wednesday, October 10, 2001 12:59 PM
I would appreciate any information about resins that have been found best for embedding of tissues for confocal microscopy. Of particular interest are lack of autofluorescence and quenching. Thank you Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
Brad, I'm not familiar with the S3500N, but most SEMs have a means of setting your record scan speed and this usually involves up to 3 different settings: 1.) Number of lines per frame (vertical rate), 2.) number of points per line (part of horizontal rate) and 3.) dwell time per point (the other part of horizontal rate) Sometimes the last two are combined in a single adjustment of time per line.
This adjustment is usually different from the viewing scan rate adjustment.
Perhaps someone who is familiar with this SEM can tell you how to make the adjustments
Ken Converse owner Quality Images third party SEM service Delta, PA
Johnson, Bradley R wrote:
} HI Ken, } I read your message, and the comment about adjusting scan speed } interests me. How would one do that? } } -Brad } } } ---------- } } From: Ken Converse } } Sent: Tuesday, October 9, 2001 3:54 PM } } To: Rick Harris; MSA, listserver } } Subject: Re: Photoshop and SEM charging } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Rick, } } My experience is that live insects don't generally charge because the } } evaporating water carries the charge away. From the looks of the image, } } I think you were using a lot of beam current. Cur that back a whole lot } } and the charging will probably go away. Also, the horizontal scan } } speed has a much greater effect on charging than the vertical speed. If } } you run a faster horizontal rate and more lines (slower vertical speed), } } this will help. } } } } Ken Converse } } owner } } Quality Images } } third party SEM service } } Delta, PA } } } } Rick Harris wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Can anyone offer any ideas on the best way to proceed with removing } } } the effects of charging on an image? I am referring to the streak } } } across the image manifested as a change in density. As an example, } } } have a look at this uncoated mite: } } } } } } http://katie.ucdavis.edu/pics/sucker2.jpg } } } } } } The nature of this projects requires that we place the live mite in } } } the scope and get a picture in the first 5 minutes. This picture was } } } taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but } } } is too poor for publication. I am looking for some help in } } } eliminating some of the charging artifacts without losing too much } } } detail. Any help will be appreciated. } } } } } } We are using PS 6 and I have the IPTK plugins. I made some progress } } } using a Gaussian filter to produce a very fuzzy duplicate image, } } } invert the image, add it to a layer and then adjust the opacity to } } } around 50%. This was only partially successful. Any help will be } } } appreciated. } } } } } } } } } } } } Rick A. Harris, Director } } } Microscopy and Imaging Facility } } } Section of Molecular and Cellular Biology } } } 1241 Life Sciences Addition } } } University of California } } } Davis, CA } } } 530 752 2914 } } } 530 754 7536 fax } } } http://katie.ucdavis.edu } } } raharris-at-ucdavis.edu } } } } } } } } } } } }
The distortion you have looks like a problem with the syncronicity of the x-y scan control and beam blanking. As the scan moves across the specimen, the beam physically moves. The electronics of the microscope "blank" the signal as the beam drops down one scan line and returns to the left hand side of the micrograph. If the blanking mechanism is slightly out of sync with the scan coils, the signal turns on while the beam is still moving from left to right. The distortion is alway at the left hand side of the micrograph and appears as a mirror image on the left and right sides of a blur. It usually is worse at faster scan rates. You may be able to get rid of the distortion by using a slower scan speed.
Cold Spring Harbor Laboratory on the north shore of Long Island, New York is seeking an experienced and responsible Biological Microscopy Facility Core Manager for the laboratory's state-of-the-art central microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, and digital imaging. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org -- Dr. David L. Spector Cold Spring Harbor Laboratory One Bungtown Road Cold Spring Harbor, New York 11724 Tel. (516) 367-8456 Fax (516) 367-8876 email: spector-at-cshl.org
Greetings Darrell, There are some good resources for learning more about multiphoton microscopy on the web. Gaining a solid conceptual understanding of the theory requires learning a little (and later, a lot) about the lasers used in this modality of optical sectioning microscopy. Here are some of the best resources I've come across:
http://swehsc.pharmacy.arizona.edu/exppath/micro/confocal.html (a great starting point for links to optical sectioning microscopy)
http://microscopy.fsu.edu/primer/techniques/fluorescence/multiphoton/multiphotonhome.html (has excellent animated renditions of the concepts)
http://www.coherentinc.com/cohrLasersAPPLICATIONS/assets/applets/CLGMPE.pdf (a PDF document from coherent, inc., a provider of lasers for multiphoton microscopy)
and you may find some helpful information and links on our site (I'm presently putting together some pertinent instructional material for our website):
http://www.itg.uiuc.edu/
If you have any specific questions, I'll be happy to try and answer them for you.
-Karl G.
_______________________________________________ Karl Garsha Specialist in Light Microscopy Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
This thread on Photoshop "enhancement" of electronic raw date brings to light an area of constant concern within my laboratory. At what point have we over "enhanced" the image, creating/altering features which potentially do not exist within the sample? It seems clear that any attempt to clone, paint, or copy/ paste fall into the category of digital artistry and should be avoid. In fact, most of the respondents have attempted to address the reduction/elimination of sample charging within the SEM, thus eliminating the need for image manipulation. Due to the highly regulated environment of the pharmaceutics industry we must adhere to strict FDA guidelines so our interpretations tend to be on the extreme side. That being said, my recommendation is to be very careful with the use of photoshop, it is a great product for "manipulating" images but our data should not be subject to artistic license. Switching directions a bit, we do employ filtering routines when analyzing data. We attempt to insert appropriate controls into the method to catch any unanticipated events. I would be very interested in the opinions of others in regards to where they draw the line on image manipulation and how they rationalize their position.
Thank you, jr
} -----Original Message----- } From: Rick Harris [SMTP:raharris-at-ucdavis.edu] } Sent: Tuesday, October 09, 2001 12:30 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Photoshop and SEM charging } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu }
I have a Nikon Labophot II and was recently told that 2.5X was the lowest mag objective that was made. However, I don't know anything about the Microphot. You can post a "want to buy" add for free on labx.com. I had a very good experience doing that a short time ago. You can also do an internet search for used microscopes. You will get a lot of hits. I have found many of the used equipment dealers to be very nice and helpful.
Judy Bowen
P.S. I think you are located at SIU Springfield. I was there from 1987-89. I was Judy Rapp then and did a little EM work. I was a friend of Donna's. Send me an e-mail at jabowen-at-buckman.com if any of this is familiar.
} } Hi, } } Our facility is looking to purchase a dry 1x objective for a Nikon Microphot } light microscope. I have contacted a few vendors and have been told that } the microscope is out of date and it will be impossible to locate such an } objective. Any help in locating this item will be appreciated. } } Thank you in advance for your help } } aruna }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donsheri695-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, October 11, 2001 at 15:06:46 ---------------------------------------------------------------------------
I assume that a good soluble organometal is easier to incorporate than a suspension of powder etc, esp for such small particles (the ruthenium red powder may be larger than your nanoparticles?), but I could easily be wrong. Sounds like you don't want to just react the stain with the particle surface before doing the study, right?
What about tetraethyl lead? Check into the catalysts (e.g. stannous octoate) used to polymerize the plga or other polymers.
I'm interested, and I would appreciate it if you could let me know what is decided upon.
Richard
Richard Thrift Richard_Thrift-at-SkyePharma.com SkyePharma, Inc 10450 Science Center Drive San Diego, CA, 92121 USA
} } } {"tbargar-at-unmc.edu"-at-sparc5.microscopy.com} 10/10/01 6:19:05 AM } } } Hi everyone,
One of our pharmacology researchers is making nanoparticles ranging in sizes from 50nm up to 250nm. The material is polylactic-acid-coglycolic-acid (PLGA). He's looking for a lipophilic electron dense particle, dye or stain that could be incorporated into the particles during their formation and which could be seen in the TEM sections. The material has to be able to be suspended or dissolved in an organic solvent.
For our part here are some of the things we've thought of using: Ruthenium Red powder, colloidal gold in 5 to 10nm range, phosphotungstic acid in ethanol.
His material will also melt at 45 degrees centigrade. So I embedded in Unicryl, UV polymerization at 4 degrees centigrade. Does anyone know if there is still heat produced during the polymerization?
This seems to be in the realm of polymer chemistry and materials science. I'd appreciate any and all help, Thanks.
Try "M & R Optical" -- Sales, Service and Calibration of all Makes and Models. The persons name is Paul Musci. He services all of our optical microscopes (all of which are older pieces of equipment).
Here is the pertinent information: Paul Musci 64 Osgood Rd. P.O. Box 515 Charlton City ,MA 01508
Tel. (508)248-5684 Fax.(508)248-4658
OR PERHAPS you could try--} A place called PYE Metallurgical Consulting, INC. One of the things this business specializes in is used and refurbished equipment. Perhaps (maybe) they could get this objective for you, or at least point you in the right direction......
This sounds reminiscent of a beta backscatter detector we used to have. Except I think we only used it for coating thickness measurements (I never used it personally). You might want to check out ASTM B-567 Standard Test Method for Measurement of Coating Thickness by the Beta Backscatter Method.
Diane Ciaburri General Dynamics Pittsfield MA
} Tony Garratt-Reed {tonygr-at-mit.edu} } 10/09/01 10:59 AM } } } Hi, Listers! } } Many years ago, when (or even before) I was an undergraduate (that would } place it in the early '60s), my aunt managed the analytical lab at a } company that produced non-ferrous metals (principally, I think, bronzes } and brasses). As one does in such situations, she invited me to tour her labs } one day. } } One of the instruments I was shown was called a "Beta Probe". A piece of } the metal was ground and polished, and was then placed face down on an } aperture in the top plate, with what I now realize was an O-ring seal. A } few buttons were pressed, and after a little while some of the electronic } counters of the day (neon tubes with 10 electrodes) began spinning round. } My memory is that the operator had some range of ratios between the } recorded numbers within which he (it was, of course, a "he" in those days, } even though my aunt was the manager) could pass the sample as meeting } specifications. } } I surmise that this was an electron milli-probe (if you'll forgive my } coining of the term!) with preset spectrometers tuned to the elements of } interest in the specification. I would very much like to find out more } about this device. Happily, my aunt is still available to be asked, but } she knows nothing of the operation of the instrument. The people who ran } it have either died, or lost touch (the company failed twenty years ago). } } If anyone can give me more information about this instrument, I would be } very interested, and most grateful. } } Thanks, } } Tony } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } * Anthony J. Garratt-Reed M.A., D.Phil. } * MIT, Room 13-1027 } * 77 Massachusetts Avenue } * Cambridge, MA 02139-4307 } * USA } * Phone: (617) 253-4622 } * Fax: (617) 258-6478 } *
{ { What about tetraethyl lead? Check into the catalysts (e.g. stannous octoate) used to polymerize the plga or other polymers. } }
I made a similar suggestion about 30 years ago, and was come down upon like a ton of bricks. Tetraethyl lead is not only extremely poisonous, it is rapidly absorbed by the skin. I understand that in companies where it is used, they have a paraffin shower, and the person is thrown under that clothes and all, no time to muck about.
Because of this, I looked up the work of J.Smid at Syracuse, NY, who uses bismuth compounds for the same purpose. From this work I learned about triphenyl bismuth, which appears to be a remarkably innocuous compound (remember the old days, when people used "bismuth" as an indigestion remedy?)
One of his published papers is:
Rawls HR, Granier RJ, Smid J, et al. Thermomechanical investigation of poly(methylmethacrylate) containing an organobismuth radiopacifying additive J BIOMED MATER RES 31 (3): 339-343 JUL 1996
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } ...
Know that you can mask this image with a "gray" and set transparency such that the image will remain unchanged. I see several places in the original image where the horizontal banding can be sampled as a row of vertical pixels which range in brightness and is representative of the charging effect. Now apply this range of grays to the gray mask. If the sampling of banded grays includes 'white', then begin with a white mask and paint the banding across the mask and apply the mask with varying transparency. Not having tried this, it is possible the opposite effect might happen ... if so, then invert the mask.
Hi, Does anyone know if there was an accessory for the WILD M5 stereo microscope that provided a (small) range of continuously variable magnification. The feature is needed to get an exact overlay of video camera images from two different microscopes.
Thanks,
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
I'd love to pick up some used copies of SEM-related texts , such as "Electron Microscopy" by Bozzola and Russell and "Scanning Electron Microscopy and X-Ray Microanalysis" by Goldstein, Newberg, Joy), or other similar textbooks. If anyone has spare copies, please contact me in private e-mail.
Does any one use any other service companies for scope service of their Philips EM400T TEM?
Other than FEI.
Joanne Crudele
-----Original Message----- } From: McFaddin, Wade [SMTP:Wade.McFaddin-at-nextekinc.com] Sent: Tuesday, October 09, 2001 7:59 AM To: Microscopy-at-sparc5.microscopy.com
Nextek Inc. is seeking a qualified individual to join our analytical laboratory team as a Materials/Failure Analysis Engineer. The ideal candidate will have a B.S. in Materials Science, Chemistry, Engineering or equivalent, and a minimum 3 years experience in the micro-electronic field; or 5 or more years hands-on lab experience in the micro-electronic materials/failure analysis field. Knowledge in cross-sectional and destructive analysis of micro-electronic assemblies and devices, and experience with analytical instrumentation such as; SEM/EDS, Micro-FTIR, Thermal Analysis, Acoustic and Optical Microscopy is a plus.
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We are going to purchase a CCD camera for our CM20 STEM. I need comment on the AltraScan 1000 made by Gatan and KeenView DUAL DOCU IMAGING SYSTEM made by SIS. Any opinion is welcome.
We have a technician job, mostly related to repair and maintenance of our scopes and other equipment. Salary is $20k to $30k per year. It will be posted next week.
Applications should be done directly to the Univ. See their job website for the posting: http://www1.umn.edu/ohr/employ.html
The basic info is as follows:
Assist in an analytical laboratory consisting of electron microscopes, Ion Beam Accelerator/spectrometer, X-ray diffraction equipment, scanning probe microscopes, optical microscopes and other instrumentation.
0 50% Daily maintenance of above mentioned instruments and associated sample preparation equipment
0 35% Troubleshooting and repair of instruments, vacuum equipment, electronics and computers
0 15% Keeping labs clean, organized and adequately supplied
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Hello, I've been trying to carbon coat some samples for SEM in a denton vacuum evaporator. I used C-rods, one thinned from the original 4mm diameter to about 1mm for a short distance (1cm) touching a flat C-rod. I manage a few times to get 12-15 nm, but often the rod breaks, or shorts out before I can get even 5 nm deposited. I like using the high vacuum on the evaporator as you get a much better coating than from our sputter coater.
Can anyone send me some pointers as to how I can get a more controlled coating with one run of the evaporator? I have to almost go to maximum power to start getting deposition registered on the monitor. I tried changing the spring so it has a more even gentler push on the sharpened rod.
It's kind of strange that you have a problem. Your setup looks like OK. I would recommend the following:
-try 5 mm instead 10 mm for 1 mm dia part. -play with spring pressure and check that spring has enough "power" when the rods will short at the end of evaporation (very usual problem). -increase current slowly with a few stops. I would suggest 10A/10-20 sec is OK. -you may try graphite or carbon. Carbon is stronger mechanically and evaporated slower and with higher current than graphite. -both rods should be CO-AXIAL. -1 mm "tip" should be sharpened to the conical shape approx 90 deg.
Personally, I prefer 6 mm carbon rods. One of them - with 1 x5 mm tip on the end:
------------------I /-------------- I {== 6 mm dia ------------------I \-------------- -1x5mm
I wish you luck. Sergey
At 03:30 PM 10/12/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
A plastic part is steam sterilized and can have a residual water drop in a small depression in the part. All the water has to be removed before the mfg process can continue. The proposed solution is to apply pressurized air (or some other gas) to remove the water drop. However, when a jet of pressurized air is applied it will blow out most of the water but leave behind small 20-50 micrometer microdroplets. These are still a problem.
I thought that I could use a dye in the test droplet of water applied to the part before it is subjected to the proposed air blast in an experimental test setup. The idea would be that if any microdroplets remained I would be able to see them under a simple macroscope. However, for such things as food dye, when it dries, the color remains and I can't tell if it is wet or dry.
One of the acceptable situation may be that most of the droplet is blown off and the remaining microdroplets dry within seconds using heat lamp, dry air, low RH in the test area, etc. So what I'm looking for is a dye that is one color when wet and either another color when dry or no color at all (clear, or white powder). My first attempts have been with cobalt chloride but driving off all the mositure is difficult. Another possibility is using a fluorescent dye that fluoresces one color when wet and has a very much reduced fluorescence when dry or better yet a different color.
I certainly appreciate you help and all suggestions will be considered, even if you have a totally different idea on how to determine when all the water is gone.
A couple of additional points: Some evaporator give a choice of voltages and this is a important feature. For metal evaporation 10V gives better control then a higher voltage, but this is not suitable for carbon rods. Graphite and even more so carbon rods, are poorer conductors than metal wire or foils, consequently evaporation is much more likely to fail at the lower voltages. Check your evaporator and if you are using something like 10 volts, then see if you can tap the transformer at a higher voltage setting, I recommend 25 or 30V. Obviously the amps required for evaporation will change. If for daily operations changing the voltage is troublesome, then I would rather put up with a touchy control for metal evaporation then with carbon evaporation failures, i.e. rather use 25V for all evaporations then 10V. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, October 13, 2001 1:50 PM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Gordon, hello } } It's kind of strange that you have a problem. Your setup looks like OK. I } would recommend the following: } } -try 5 mm instead 10 mm for 1 mm dia part. } -play with spring pressure and check that spring has enough "power" when } the rods will short at the end of evaporation (very usual problem). } -increase current slowly with a few stops. I would suggest 10A/10-20 sec } is OK. } -you may try graphite or carbon. Carbon is stronger mechanically and } evaporated slower and with higher current than graphite. } -both rods should be CO-AXIAL. } -1 mm "tip" should be sharpened to the conical shape approx 90 deg. } } Personally, I prefer 6 mm carbon rods. One of them - with 1 x5 mm tip on } the end: } } ------------------I /-------------- } I {== 6 mm dia } ------------------I \-------------- } -1x5mm } } I wish you luck. Sergey } } } At 03:30 PM 10/12/01 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello, } } I've been trying to carbon coat some samples for SEM in a denton vacuum } } evaporator. I used C-rods, one thinned from the original 4mm diameter to } } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a } } few times to get 12-15 nm, but often the rod breaks, or shorts out before } } I can get even 5 nm deposited. I like using the high vacuum on the } } evaporator as you get a much better coating than from our sputter coater. } } } } Can anyone send me some pointers as to how I can get a more controlled } } coating with one run of the evaporator? I have to almost go to maximum } } power to start getting deposition registered on the monitor. I tried } } changing the spring so it has a more even gentler push on the sharpened } } rod. } } } } The rods I use after sharpening look like this: } } } } --------| |--------------- } } | | } } ---------| } } ---------| } } | | } } --------| |-------------- } } } } Hopefully the ascii art comes out in email. } } } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } } \ } } Gordon Ante Vrdoljak Electron Microscope } } Lab } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } phone (510) 642-2085 Berkeley CA 94720-3330 } } fax (510) 643-6207 cell (510) 290-6793 } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } }
I use the same method, and I have the same problem if I thin the rods down too much. Try turning it down to taper down to 2mm over 6mm of length, and make sure that the flat end of the other electrode doesn't have any debris left on it from previous use. For the flat-ended rod, I use 6.5mm diameter (I don't know whether it is graphite or carbon, I don't think it matters for this one).
In contrast to Sergey's experience, I have found that graphite needs to get much hotter to evaporate than does carbon, and the holder assembly all became red hot when I tried graphite.
Jim Darley at Proscitech sells both types.
My only relationship with him is as a satisfied customer.
good luck
rtch
} } Hello, } I've been trying to carbon coat some samples for SEM in a denton } vacuum evaporator. I used C-rods, one thinned from the original 4mm } diameter to about 1mm for a short distance (1cm) touching a flat } C-rod. I manage a few times to get 12-15 nm, but often the rod } breaks, or shorts out before I can get even 5 nm deposited. I like } using the high vacuum on the evaporator as you get a much better } coating than from our sputter coater. } } Can anyone send me some pointers as to how I can get a more } controlled coating with one run of the evaporator? I have to almost } go to maximum power to start getting deposition registered on the } monitor. I tried changing the spring so it has a more even gentler } push on the sharpened rod. }
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
We would like to upgrade our PC containing SemAfore 3.02 hooked with JEOL JSM5300. The old card is using the ISA however all the new boards do not have the ISA but PCI only. So we've asked JEOL people to get ISA extender. After getting the new 1 Ghz PC, hooking the ISA extender, hooking the card, got everything hooked up ... I think the engineer told me ... "very-very-very low signal". So we looked around and found one motherboard with one ISA and the rest PCI. Hooked everything up. Still the same thing happened. The engineer told us that we need to purchase the USB type which costs a hefty sum in order to use with the new PC. So now we are back to the old PC. My worry is what happen when the old PC konk.
Any suggestion or somebody experienced this? btw, the engineer also tried with SemAfore 4.0 and it did not work. } } Regards, } } Fauzi } Malaysian Rubber Board }
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Gordon, The most obvious thing, that no one has mentioned yet, is that the non-sharpened rod should be shaped at a 45 degree angle with the face towards your sample. The one mm rod should hit roughly in the center of the 45 degree face. This allows a better distribution of carbon towards your sample. 1 cm of 1mm rod seems awfully long to me. The trade-off is distance from sample vs heating of sample. Closer gives you more rapid coating due to the larger solid angle. Too close gives very intense heating. Too far away can also impart too much heat because it can take so long to get the thickness you're looking for. My understanding is that 9-10 cm should be a good distance. Someone with more math skill than I can probably tell you how many mm of 1mm diameter carbon you need at that distance to get 5, 10 or 15 nm coating.
Ken Converse owner Quality Images third party SEM service Delta, PA
Gordon Vrololjak wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I've been trying to carbon coat some samples for SEM in a denton vacuum } evaporator. I used C-rods, one thinned from the original 4mm diameter to } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a } few times to get 12-15 nm, but often the rod breaks, or shorts out before } I can get even 5 nm deposited. I like using the high vacuum on the } evaporator as you get a much better coating than from our sputter coater. } } Can anyone send me some pointers as to how I can get a more controlled } coating with one run of the evaporator? I have to almost go to maximum } power to start getting deposition registered on the monitor. I tried } changing the spring so it has a more even gentler push on the sharpened } rod. } } The rods I use after sharpening look like this: } } --------| |--------------- } | | } ---------| } ---------| } | | } --------| |-------------- } } Hopefully the ascii art comes out in email. } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } } }
Two postdoctoral positions (see below) are available immediately. If interested, send a CV including the names of referees to ldm3-at-apollo.numis.nwu.edu
1. Quantitative Electron Microscopy
A postdoctoral position is available in the area of quantitative TEM/TED of materials starting immediately. The research project focuses on developing quantitative methods of solving structures from diffraction patterns and HREM images. Analysis of diffraction patterns will be via Direct Methods to recover estimates of the phases; analysis of HREM images will be to develop methods to routinely perform translational/rotational averaging. The work will involve collaboration with scientists at LBL and elsewhere. A good background in computer programming in fortran or C is required, as well as an understanding of dynamical diffraction theory. Prior experience in HREM or Direct Methods would be an advantage.
2. Surface Microscopy
The position would involve work using the unique HREM/Surface Science facility at Northwestern University (see http://www.numis.nwu.edu). The primary research area will be metalic nanostructures on semiconductor surfaces, although elements of the work will also overlap with other ongoing projects involving magnetron deposition of quasicrystalline thin films, hard coatings and oxide surface structures. A strong background in basic TEM techniques as well as some familiarity with thin film growth, surface science and other characterization techniques such as XPS are important.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
I have used approx. 20 different Denton 502's for evaporation (and FE) over the last 30 years. I also recondition and sell them. Since you are maybe using one of them "carbon fiber Flash Evaporators"? sputter coaters, the speed to evaporate in HV has to be SLOW. Have sufficient spring pressure, s l o w l y bring up the current, watch the rod (with welder's glasses or cover the bright zone by viewing over the holder bar, DONT LOOK INTO THE BRIGHT SPOT!! Of course you know that!) until some fine sparks appear, immediately reduce the current below sparking and watch your indicator for appropriate coating. It works 99.9% of the times. Regards, Markus F. Meyenhofer Microscopy Labs
Colleagues, can somebody recommend (by own positive experience!) an antibody against collagen, e.g. Type IV-Collagen ? Antibody should work in Immuno-EM, preferably tolerating glutaraldehyde fixed material.
Thanks for direct short, informal e-mail to me.
Peter
************************************************** please reply to: peter.heimann-at-biologie.uni-bielefeld.de ("E-MAIL")
Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : xx49(0)521-106-5654 WEB-Site: http://www.uni-bielefeld.de/biologie/Entwicklungsbiologie WEB-Site: http://www.uni-bielefeld.de/SFB549 ******************************************************
Dear Gordon, I have a very old JEOL evaporator and what I found was: 1. I sharpen my 5 mm dia. rods to 1 mm for no more than 4 to 5 mm. Flatten the rod it rests on with the edge of a glass slide. 2. Use pressed carbon, not graphite. It heats more readily by resistance. 3. Carefully align and tighten the rods before you let the spring-loaded one rest against the center of the fixed one. This avoids putting a twist in your thinned, weak rod. 4. Don't knock the rods as you are putting the lid on. I find you have to turn up the current slowly, watching through welding goggles, just until the tip starts to spit sparks. Leave it there and it will climb up by itself after that. At 03:30 PM 10/12/01 -0700, you wrote: } Hello, } I've been trying to carbon coat some samples for SEM in a denton vacuum } evaporator. I used C-rods, one thinned from the original 4mm diameter to } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a } few times to get 12-15 nm, but often the rod breaks, or shorts out before } I can get even 5 nm deposited. I like using the high vacuum on the } evaporator as you get a much better coating than from our sputter coater. } } Can anyone send me some pointers as to how I can get a more controlled } coating with one run of the evaporator? I have to almost go to maximum } power to start getting deposition registered on the monitor. I tried } changing the spring so it has a more even gentler push on the sharpened } rod. } } The rods I use after sharpening look like this: } } --------| |--------------- } | | } ---------| } ---------| } | | } --------| |-------------- } } Hopefully the ascii art comes out in email. } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab I hope this helps. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
The DTSA manual describes the method of "smoothing" a spectrum as a 5, 7, or 9 point Savitsky-Golay polynomial. Does anyone know how this is applied. Is it similar to passing a filter through the data with weighted coefficients? I would appreciate if anyone can give me the algorithm. Thanks in advance.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
In a message dated 10/15/01 2:27:04 PM, walck-at-ppg.com writes:
} The DTSA manual describes the method of "smoothing" a spectrum as a 5, } 7, or 9 point Savitsky-Golay polynomial. Does anyone know how this is } applied. Is it similar to passing a filter through the data with weighted } coefficients? I would appreciate if anyone can give me the algorithm. } Thanks in advance
That is exactly what it is. The S&G coefficients allow fitting polynomials of various degrees (e.g. a quadratic polynomial) to data in order to smooth it. The 7 point coefficients for a quadratic, for example, are -.0952 .1429 .2857 .3333 .2857 .1429 -.0952 to be multiplied by each group of the histogram values in order to produce one new point at the center of the 7. This is then repeated at every point to generate a new, smoother curve.
I would like to announce the debut of the Food Structure and Functionality Discussion group at the AOCS (Americal Oil Chemists Society) website, webaddress:
http://www.aocs.org/ubbcgi/ultimatebb.cgi
This discussion group is open to all agri-food researchers from Government, Industry and universities to discuss topics of interest, problems, sample preparation, etc. much like the function of this Microscopy listserver, as well as being a source of information for the activities of the Food Structure and Functionality Forum - a division of AOCS, and its yearly Symposium.
We want to make it an interesting and valuable resource for anyone who visits it. Please visit the site today and start or join a discussion! The site is up and ready for you to use.
Kind regards,
Paula Allan-Wojtas, Chair Food Structure and Functionality Forum - a division of AOCS
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
I was going to take a look at your book and see what they were across the center row for an image Gaussian smoothing filter if nobody replied. Somebody is currently borrowing my copy however. (Fred, if you are reading this, it is time to return it.)
Let me bother you with another question and I made it public on the Listserver. How do you handle the ends of the data array, i.e. in a 1024 array, the indices 1-3 and 1022-1024. When you apply the coefficients, they sum to one, so you can't just use some of them.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Monday, October 15, 2001 2:40 PM To: walck-at-ppg.com; microscopy-at-sparc5.microscopy.com
In a message dated 10/15/01 3:49:53 PM, walck-at-ppg.com writes:
} I was going to take a look at your book and see what they were across the center row for an image Gaussian } smoothing filter if nobody replied. Somebody is currently borrowing my copy however. (Fred, if you are } reading this, it is time to return it.)
Yes, they are in the book. Tell him to buy his own copy!
} Let me bother you with another question and I made it public on the Listserver. } How do you handle the ends of the data array, i.e. in a 1024 array, the } indices 1-3 and 1022-1024. When you apply the coefficients, they sum to } one, so you can't just use some of them.
Edges are always a problem. The simplest solution is to consider them a mirror, so that the -1, -2, -3 positions have the same values as +1, +2, +3 (and the same thing at the other end). The other common choices are to perform some kind of extrapolation on the data at each end (the simplest of which is linear) or to make your addressing wrap around (i.e., -1 = 1023, -2 = 1022, etc.) which is not appropriate for most cases.
In any case, you can expect to lose some data at the ends (or for images, the edges).
Fellow microscopists I have a two dye sublimation printers that are gathering dust in my lab. If anyone is interested, you can make me an offer. The the first printer circa 1994 Seiko ColorPoint professional 300 dpi dye sublimation unit with an interface box to allow screen captures from a computer monitor. The unit includes a spare ribbon and 50 sheets of paper. The second is a Fargo primera pro elite 300 dpi resolution. This unit is a win printer, and will only work with Windows 95 or 98 (which I no longer use). The reason I'm not using them is that they simply cost too much to run. I replaced both of them with a networked inkjet printer that is much cheaper to run and suits my requirements just as well.
Both include manuals and are for sale as is. If anyone is interested they can drop me a note. -- Glenn
=============================================================================== Glenn Poirier 3450 University St, rm. 238 MicroAnalytical Laboratory Montreal, Qc Earth and Planetary Sciences tel (514) 398 6774 McGill University fax (514) 398 4680 email: glennp-at-eps.mcgill.ca http://castaing.eps.mcgill.ca
++ Millenium hand and shrimp ++ ===============================================================================
A physicist colleague asked if I knew how to make microscope slides repellent so that dust and the paper fibres he is studying would not adhere to them. Apart from applying a thin coat of some non-polar material, I didn't have any useful suggestions - I'm usually trying to make things stick, not vice versa.
Anyone have a more useful and/or more specific suggestion or protocol?
Thanks, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
It depends what you want: if you need to prevent ionic interactions in the solution (like ion-pairs) make glass hydrophobic by siliconization. If you want to remove static electricity (which is usually attracts the dust) you may have to make glass electro-conductive (make it wet in simply case, poly-lysine may work as well). Both recommendations are quite controversial because siliconization make glass hydrophobic and it increase the chance of electization when dry. I would suggest that there is no solution for your case: dust is heterogeneous and some part of this nasty thing will find the way to contaminate your sample. In this battle the dust is always a winner.
Paper fibers, cellulose, has a lot of -OHs, so the nature of adhesion is polar/ion (in the water solution). Siliconization would help.
Sergey
At 09:26 AM 10/16/01 +1000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
on 10/16/01 7:26 AM, Rosemary White at Rosemary.White-at-pi.csiro.au wrote:
} } A physicist colleague asked if I knew how to make microscope slides } repellent so that dust and the paper fibres he is studying would not adhere } to them. Apart from applying a thin coat of some non-polar material, I } didn't have any useful suggestions - I'm usually trying to make things } stick, not vice versa. } } Anyone have a more useful and/or more specific suggestion or protocol? } Dear Rosemary, I don't know whether this will work for the particular application, but many scientific supply houses make a silicone solution designed to make glass hydrophobic. I think Sigma's is called Siliclad. Good luck to a fellow physicist. Yours, Bill Tivol
Same situation here - we usually want things to stick - but poly-lysine is out. What charge do the particles have - maybe there is something with the same charge that would repel?
My other thought is PTFE/Teflon in it various presentations ("thin coat of some non-polar material"). I use it on the moving parts of my bicycle as a solution in spray form and also applied as a liquid solution. I can't remember what the solvent/solvent mixture is but I would guess that it is alcohol based. I have never tried it on microscope slides but is might work.
Would it do the job?
Med vänliga hälsningar/With best regards
Gareth
Då du älskar, älskar med glödande hetta. Då du hatar, hata i flammande blixtar. Då du festar, festa som om vardagen inte fanns. Då du går, gå fort, men lämna alltid dörren på glänt......
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
Hi I would like to thank everybody very much for your advice on the subject of { fluorescence microscopy and TEM on same sample} . I am working on the procedure and may come back with further questions.
Glass microscope slides can be made hydrophobic using hexamethyldisilAzane (HMDS) (Aldrich Chemical 37,921-2). Simply place the separated slides in a container like a wide mouth glass jar with an aluminum or Teflon lined cap. Add a few drops of HMDS and it will vaporize overnight in the sealed jar and react with the silanols on the surface. I prefer to heat the jar in an oven at 70° overnight to drive off residual ammonia. Leave the jar slightly cracked open. The next day, in a hood, open and remove the slides. They will now bead water. This will be a molecular level coating and invisible.
Things like Rainex® and Aquapels® will work but can't be applied in the vapor state. You do not want to create artifacts (streaks) on the slides, so use the vapor state application of hexamethyldiSILAZANE (HMDS). HexamethylsilOXane is NOT the same material. HMDS is simple, easy to apply, and safely handled.
Use gloves, a hood, and respirator when handling these materials. Let the dropper and the opened jar with HMDS evaporate overnight in the hood and then place them in the trash the next day. Do not breath HMDS vapors at all! Read the MSDS information on handling.
I had a 'never opened bottle and wax sealed bottle' of chlorosilane spontaneously explode in a closed cabinet. It formed lots of white smoke and HCl. Nobody was around at the time it happened. This will make you a believer in using HMDS when possible.
These are my opinions only but based on my experiences.
Paul Beauregard Senior Research Associate Monroeville, PA 15601
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-pi.csiro.au] Sent: Monday, October 15, 2001 7:26 PM To: Microscopy-at-sparc5.microscopy.com
Dear all,
A physicist colleague asked if I knew how to make microscope slides repellent so that dust and the paper fibres he is studying would not adhere to them. Apart from applying a thin coat of some non-polar material, I didn't have any useful suggestions - I'm usually trying to make things stick, not vice versa.
Anyone have a more useful and/or more specific suggestion or protocol?
Thanks, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Wow! is this ancient history! My first paper ever was about the determination of the thickness of carbon films correlated with optical density. Which meant that I did a _lot_ of films, with different length carbon rods (using an old Kinney evaporator actually). The primary result was that OD and thickness have a linear relationship from about 5nm to 100nm. (And right now I am having a senior moment and not remembering the exact correlation--and where is that paper now that I need it???) On the practical side, 1mm carbon rod would give a 10nm film, and as I remember the correlation was fairly linear between 0.5mm (5nm) up to 5mm (50nm). I simply use the old "folded filter paper" method for evaluation, knowing that an OD of about 1 gives a very heavy film--probably about 100nm. As Ken says, geometry has a lot to do with the final thickness deposited, but once you work it out for your system it should be fairly consistent. If my brain functions and I can get some of these tired neurons to fire (for more precise information), I'll forward it along. Roger
On Mon, 15 Oct 2001 08:03:44 -0400, Ken Converse 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 | -----------------------------------------------------------------------. | | | Gordon, | The most obvious thing, that no one has mentioned yet, is that the | non-sharpened rod should be shaped at a 45 degree angle with the face | towards your sample. The one mm rod should hit roughly in the center | of the 45 degree face. This allows a better distribution of carbon | towards your sample. 1 cm of 1mm rod seems awfully long to me. The | trade-off is distance from sample vs heating of sample. Closer gives | you more rapid coating due to the larger solid angle. Too close gives | very intense heating. Too far away can also impart too much heat | because it can take so long to get the thickness you're looking for. My | understanding is that 9-10 cm should be a good distance. Someone with | more math skill than I can probably tell you how many mm of 1mm diameter | carbon you need at that distance to get 5, 10 or 15 nm coating. | | Ken Converse | owner | Quality Images | third party SEM service | Delta, PA | | Gordon Vrololjak wrote: | | } ------------------------------------------------------------------------ | } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
| } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | } -----------------------------------------------------------------------. | } | } | } Hello, | } I've been trying to carbon coat some samples for SEM in a denton vacuum | } evaporator. I used C-rods, one thinned from the original 4mm diameter to | } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a | } few times to get 12-15 nm, but often the rod breaks, or shorts out before | } I can get even 5 nm deposited. I like using the high vacuum on the | } evaporator as you get a much better coating than from our sputter coater. | } | } Can anyone send me some pointers as to how I can get a more controlled | } coating with one run of the evaporator? I have to almost go to maximum | } power to start getting deposition registered on the monitor. I tried | } changing the spring so it has a more even gentler push on the sharpened | } rod. | } | } The rods I use after sharpening look like this: | } | } --------| |--------------- | } | | | } ---------| | } ---------| | } | | | } --------| |-------------- | } | } Hopefully the ascii art comes out in email. | } | } | } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ | } Gordon Ante Vrdoljak Electron Microscope Lab | } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall | } gvrdolja-at-nature.berkeley.edu UC Berkeley | } phone (510) 642-2085 Berkeley CA 94720-3330 | } fax (510) 643-6207 cell (510) 290-6793 | } | } | } | } | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
A local research physician's widow has contacted me to find out whether her husband's microscope has any value or is still useful. It's a Nikon model L-Ke (possibly with a camera attachment). Can anyone help with information?
Many thanks, Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Just wanted to confirm our schedule and request driving instructions/hotel info for the Hitachi site. Also wondering which airport is best to fly into (?).
I need to produce clear cut high resolution pictures of the peptidoglycan layer of E. coli. I am looking for a staining method that shows some specificity for peptidoglycan (may be some method for polysaccharides, I guess) while allowing high resolution immages.
Preferably the method should work with Epon-Araldite embedded material fixed in glutaraldehyde and osmium.
Does anyone have suggestions for such a reliable stainning method?.
Dr. A.P. Alves de Matos Curry Cabral Hospital Lisbon apmatos-at-ip.pt
We were wondering if anyone else has run into this problem: we bought a Hitachi VP S-3000N SEM. They offer a 125 mm multiple specimen holder - essentially an aluminum disc on a small column that has 33 holes in it to hold 33 stubs. We bought it and have now discovered that none of the commercial vendors sell pin-type stubs that will actually fit into these holes! The majority of stubs have pins that are 3mm in diameter. The holes are only 2 mm in diameter. EMS can make the stubs at 13 cents a piece but the minimum order is 100,000. Has anyone out there found a source of stubs at a reasonable price (Hitachi offered to sell them at approximately $25.00 a piece!) or are people just drilling the holes in the holder to make them bigger? I'd really like to hear how other people are handling this!
Lesley
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
I made my own multi stub holder. But if I had what you have I'd head down to the shop and drill/ream out those holes or have a competent shop guy do it. Maybe 20-30 minutes of work, tops.
} We were wondering if anyone else has run into this problem: we bought a } Hitachi VP S-3000N SEM. They offer a 125 mm multiple specimen holder - } essentially an aluminum disc on a small column that has 33 holes in it to } hold 33 stubs. We bought it and have now discovered that none of the } commercial vendors sell pin-type stubs that will actually fit into these } holes! The majority of stubs have pins that are 3mm in diameter. The } holes are only 2 mm in diameter. EMS can make the stubs at 13 cents a } piece but the minimum order is 100,000. Has anyone out there found a } source of stubs at a reasonable price (Hitachi offered to sell them at } approximately $25.00 a piece!) or are people just drilling the holes in } the holder to make them bigger? I'd really like to hear how other people } are handling this! } }
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Applications for an NEI-supported post-doctoral position to study M cells in the mammalian conjunctiva using LM & EM techniques are being accepted.
M cells are classically found overlying the mucosal lymphoid follicles (Peyer's Patches) of the intestinal tract. The importance of M cells in the mucosal immune response has only recently been appreciated. M cells have a distinctive morphology that includes an invagination of their basolateral membrane to form a "pocket" filled with macrophages, lymphocytes and other antigen presenting cells. The apical portion of the M cell forms thin bridges that separate the extracellular world from the underlying lymphoid cells. This narrow bridge can result in the distance from the apical membrane to the underlying pocket of less than 2 microns. The M cell acts as an antigen sampling cell by capturing souble and particulate lumenal antigens, transcytosing them across the thin cytoplasmic bridge) and releasing them into the subcellular pocket filled with macrophages and other antigen presenting cells. Endocytosis of antigens by M cells is thought to be the first step in generating a mucosal immune response. There is considerable interest in the development of immunogens targeted to M cells to develop immunity against diseases such as cholera. It has also been realized that opportunistic pathogens (e.g., HIV, Salmonella, Shigella) selectively bind to intestinal M cells and exploit the transcytosis process to allow them to penetrate the epithelial barrier.
M cells have also been found to occur in tonsils, bronchi, and the nasal cavities. The presence of M cells in the ocular conjunctiva is much more controversial. In collaboration with Dr. Cecil Moore, DVM (a veterinary ophthalmologist and chair of Veterinary Medicine & Surgery, MU Veterinary School), my lab has recently found LM, TEM, and SEM evidence of cells with the highly distinctive morphology of M cells in the conjunctiva of dogs (paper under review). Another group found similar views in the Guinea pig. The bulk of the literature claims M cells don't exist in the eye. This is mostly because no one has ever tested the ability of follicular associated epithelium in the eye to selectively transport antigens. Our new grant is designed to test the ability of these putative M cells to selectively bind and transport antigens of pathological importance. We intend to screen the conjunctiva of multiple mammalian species to prove the widespread existence of conjunctival M cells. We will also be testing whether ocular immunization is superior to immunization at other sites in regards to generating an ocular immune response. Candidates should have experience in light and/or electron microscopy.
The project is mostly LM, TEM and SEM with some immunology later on. Funding for the postdoc is $28,260 plus full benefits. There is funding for 3 years but departmental policy requires offers to be made for 1 year with re-appointment for subsequent years dependent on satisfactory progress. The postdoc is available immediately but I would be willing to wait a reasonable amount of time for the right candidate. Send CV and names of three references by e-mail to 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 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I am seeking feedback--good and bad--about a planned x-ray detector system purchase.
I'm doing semiconductor work and need light element detection (down to B). I do not have LN2. As a result, I've narrowed the candidates to Noran's cryo cooled 10mm detector and their Quest software system. The detector would be mounted on an Amray 1910 FESEM.
The Noran system comes with its own PC. I'd rather have it integrated into my SEM control PC. But I suppose this would work out OK in the long run. Noran uses a Compaq PC with dual Ethernet ports. One port talks to the pulse counter and detector interface. I may try to use the other Enet port to connect the SEM PC.
I would appreciate any feedback from current users of this product. I'm wondering about reliability and up-time. Also, how are these systems warranted and maintained? Off-list responses are welcomed.
Dear Listers I am presently undetaking an Honours degree at La Trobe University, Victoria, Australia.I have been researching flower pollen using a Scanning Electron and Confocal microscopes. I am particularly interested to hear your views on the following question: Do you see a link between science and art? If so, how and why? Thank you for considering my question. Yours sincerely Judi Bowden La Trobe University Victoria Australia
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (j.miyan-at-umist.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, October 16, 2001 at 12:07:24 ---------------------------------------------------------------------------
Email: j.miyan-at-umist.ac.uk Name: Jaleel Miyan
Organization: UMIST
Education: Graduate College
Location: Manchester, UK
Question: We are looking for a method to stain for proteoglycans in sections of rat brain. I had heard that you can use a critical eletrolyte method using alcian blue and varying magnesium concentrations.
Can you direct me to a protocol and/or comment on the accuracy of this and any other possible method.
This is a reply to all the people who have been asking me about what kind of success I have had with the cell wall dyes.
First of all, I am trying to stain 1 micron thick sections of LR White-embedded cotyledons of Vicia faba. I got really nice staining with a 1% solution (I know - it's pretty strong!) for 2 min (no heating). I washed the sections after staining in running water for 15 min to remove any unbound stain. The stain instantly got weaker when excited (green filter) if I mounted the sections in water or 50% glycerol. So I mounted them in Zeiss immersion oil, which gave me a really, really nice clear image. The only problem was that, as was pointed out by Susan Joa, the dye binds to 1-4 Beta Glucans, so the starch grains were fluorescing pretty strongly. However, in my opinion the image I got was much better than any calcofluor image I have ever taken. In addition, the nuceli were lightly stained (does anyone know why this is?).
Hi: We are looking (again) for a used Zeiss 10 TEM for the Los Angeles area. Please let me know if you have one for sale or if you know of somebody selling. Thank you, Peter Jordan 909 302-9130
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mryder-at-brookes.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, October 17, 2001 at 05:31:30 ---------------------------------------------------------------------------
Question: Why do my wholemount lowish power root photographs sometimes come out yellow, on the same film, with the same lighting. Sometimes the same root taken a few minutes later will be yellow when it was the usual white on a previous frame.
After my company bought a 3000N, I recognized that the Hitachi sample holder system was relatively expensive. So I designed a 50 mm holder with space for 8 12-13 mm dia. relatively inexpensive Zeiss stubs and had a local machine shop make 7 of them. That included set screws for the sample stubs and an appropriate center hole to screw in a Hitachi base. The 3000N memory system was used to remember the centerpoint of each of the 8 sample sites so moving from sample to sample is easy. The cost was easily recovered considering the high cost of the Hitachi mount system. All of the users were assigned a holder and it's worked without a hitch ever since.
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We were wondering if anyone else has run into this problem: we bought a Hitachi VP S-3000N SEM. They offer a 125 mm multiple specimen holder - essentially an aluminum disc on a small column that has 33 holes in it to hold 33 stubs. We bought it and have now discovered that none of the commercial vendors sell pin-type stubs that will actually fit into these holes! The majority of stubs have pins that are 3mm in diameter. The holes are only 2 mm in diameter. EMS can make the stubs at 13 cents a piece but the minimum order is 100,000. Has anyone out there found a source of stubs at a reasonable price (Hitachi offered to sell them at approximately $25.00 a piece!) or are people just drilling the holes in the holder to make them bigger? I'd really like to hear how other people are handling this!
Lesley
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
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Dear Judy,
While I'm not a microscopist, I think there is a strong link in that some of the work one does for science can certainly become art.
If you are interested in further exploring the connection, I suggest you contact David Scharf of Los Angeles CA. His email address is dscharf-at-pacbell.net. David is an Artist who produces color SEM photomicrographs and his work is outstanding. In fact, he recently won an EMMY award for his work on a National Geographic documentary film called "Body Snatchers".
Sincerely,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: "01151938-at-mrc.vic.edu.au"-at-sparc5.microscopy.com [mailto:"01151938-at-mrc.vic.edu.au"-at-sparc5.microscopy.com] Sent: Tuesday, October 16, 2001 7:10 PM To: Microscopy-at-sparc5.microscopy.com
Dear Listers I am presently undetaking an Honours degree at La Trobe University, Victoria, Australia.I have been researching flower pollen using a Scanning Electron and Confocal microscopes. I am particularly interested to hear your views on the following question: Do you see a link between science and art? If so, how and why? Thank you for considering my question. Yours sincerely Judi Bowden La Trobe University Victoria Australia
Is there a procedure for keeping a pellet of cells together (as an intact pellet) while processing it for thin sectioning? I am working with virus infected cells removed from the substrate via trypsin and desire to keep the pellet densely packed and intact for subsequent processing steps. The embedded pellet will then be thin sectioned. Thanks in advance.
Tom Doman Animal Diagnostic Laboratory Penn State University
I know they sell epi-fluorescence attachments for stereoscopes but I am wondering how good they are in a practical sense. I want to detect clusters of FITC-tagged bacteria clustered on large pieces of mucosa measuring approximately 2 x 4 cm. Does anyone out there have any experience in this type of work. TIA, tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} } } Email: j.miyan-at-umist.ac.uk } Name: Jaleel Miyan } } Organization: UMIST } } Education: Graduate College } } Location: Manchester, UK } } Question: We are looking for a method to stain for proteoglycans in } sections of rat brain. I had heard that you can use a critical } eletrolyte method using alcian blue and varying magnesium } concentrations. } } Can you direct me to a protocol and/or comment on the accuracy of } this and any other possible method. } } MAny thanks } } Jaleel } } --------------------------------------------------------------------------- Jaleel There are a variety of cationic dyes, which because of their charge, will bind to proteoglycans, and that are electron dense. Many moons ago, I used alcian blue, ruthenium red, safranin O, and cetyl-pyridinium chloride (?) to contrast the ECM in heart muscle (see Robinson, et al, lab. Invest. 49(4):482-97 1983.) Also, in the 1970's and 1980 the Simionescu's had a lot of publications using various cationic dyes.
good luck, Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
You can resuspend your cell pellet in a few drops of 22 percent bovine albumin prior to glutaraldehyde fixation. Then, when you add the glut., the BSA proteins will crosslink and become a solid chunk which can then be processed as a piece of tissue.
An alternate method that I use is to resuspend the pellet and then centrifuge at each step of the prep. I routinely process virally infected cell monolayers this way. I like to embed in Spurrs rather than Epon because of its lower viscosity, allowing for easier pelleting. I do not use trypsin to lift the cells. Rather, I fix and post-fix the cells right in the culture dishes. Then, after the osmium rinses, I scrape the monolayers off of the dishes and pellet them in Eppendorf tubes for the remainder of the process.
I hope this helps!
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Wed, 17 Oct 2001, Tom Doman 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 Colleges, } } Is there a procedure for keeping a pellet of cells together (as } an intact pellet) while processing it for thin sectioning? I am working } with virus infected cells removed from the substrate via trypsin and } desire to keep the pellet densely packed and intact for subsequent } processing steps. The embedded pellet will then be thin sectioned. } Thanks in advance. } } Tom Doman } Animal Diagnostic Laboratory } Penn State University }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleges, } } Is there a procedure for keeping a pellet of cells together (as } an intact pellet) while processing it for thin sectioning? I am working } with virus infected cells removed from the substrate via trypsin and } desire to keep the pellet densely packed and intact for subsequent } processing steps. The embedded pellet will then be thin sectioned. } Thanks in advance. } } Tom Doman } Animal Diagnostic Laboratory } Penn State University
After fixing, pellet it down in the buffer in an eppendorf tube remove the buffer and add a drop of low melting point agarose made up in buffer (add no more than the same size of pellet) use a warmed pin (37 degrees) or long needle to swirl the pellet into the agarose before it sets. When set use a razor blade to cut off the end of the eppendorf to get at the pellet poke out the pellet onto a drop of buffer on dental wax or parafilm chop the agarose pellet into 1 mm squares put into a scintillation vial and process as normal for tissue pieces.
If the layer of cells is almost invisible, cover with a tiny amount of low melting point agarose set by cooling finish the processing in the eppendorf as long as the low melting point agarose does not lift off the bottom of the tube but traps the cells in place. In this case the thickness of the LMP agarose should allow penetration of the chemicals so should be very thin.
Elaine
-- Dr. Elaine Humphrey Director, Biosciences Electron Microscopy Facility First Vice President, Microscopy Society of Canada 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 website: www.emlab.ubc.ca
Remove the cells from the substrate (if you're dealing with an enveloped virus, you might want to use means other than trypsin).
Pellet cells gently (2000 x g for 10 min)
Resuspend in glut in microfuge tube.
Swirl for 30 sec, then pellet again.
Let pellet sit without resuspending to fix. Time will depend on pellet size.
After 1 to several hours, remove fix with pipet; cut end off plastic tube with razor blade. Holding tip with forceps under dissecting, scope scoop out pellet with flattened and pointed applicator stick.
Dry further with pie-shapped wedge of filter paper by touching only the tip to the cell mound. The consistency should be that of cooked oatmeal. If the pellet is not dry enough, cells will disperse in the agar and be hard to find in the EM. If it is too dry, ultrastructure will suffer.
Divide the pellet into clumps about 1 mm cubed with a razor blade or spatula and surround them with molten agar*.
Let harden, and wash in buffer. Proceed with osmium, etc., as though you had tissue blocks.
*Mix 1 g purified agar with 100 ml water (or 0.1 g in 10 ml); melt by boiling--watch out, it easily boils over. Put into small tubes; cap or cover with Parafilm. Store at 4 degrees C. When needed, melt in beaker of boiling water. Let cool till barely warm to touch--don't cook the cells. (Agar solidifies at about 36 degrees and melts at about 95 degrees.)
Do not put agar-encased cells into glut. It will cross-link the agar so that subsequent solutions don't penetrate. However the remaining glut in the (unwashed) cells prior to encasing in agar won't hurt.
Good luck.
Sara Miller
On Wed, 17 Oct 2001, Tom Doman wrote:
} Date: Wed, 17 Oct 2001 10:14:22 -0500 } From: Tom Doman {jtd1-at-psu.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Cell pellet processing for TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleges, } } Is there a procedure for keeping a pellet of cells together (as } an intact pellet) while processing it for thin sectioning? I am working } with virus infected cells removed from the substrate via trypsin and } desire to keep the pellet densely packed and intact for subsequent } processing steps. The embedded pellet will then be thin sectioned. } Thanks in advance. } } Tom Doman } Animal Diagnostic Laboratory } Penn State University } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts dropping. The problem developed over time. In the beginning, it would drop by very little and go back up again. Later on, it started dropping by a lot more and it took longer to get back up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing this problem? Any tips or suggestions? We appreciate the help and thank you all in advance.
I am a little rusty on this, but when I did my masters in interdisciplinary arts, I used the SEM. The connection I used between science and art was creativity. There are many fields that we can be creative in...science and art are two of them. We consider art the mother of creativity. Yet, we live in a scientific/technological paradigm these days more than a religious paradigm like the Middle Ages. Those main religious and scientific paradigms overlap(think of interlapping rings much like the Olympic rings) into other disciplines like medicine, art, history, and law. The thread that keeps it all woven together is creativity. There are many definitions to clarify, but I believe looking at science and art as part of a bigger construct fueled by creative energy will help you with the larger picture. The more immediate picture like what is science and what is art and what is creative remains for us to define and redefine. How we define it is by the tools we use, the message we seek, the role of transformation and discovery in what we produce. Good Luck!
We do this all the time with cell suspensions. Pellet the live cells (~1,000 rpm for 10 minutes), carefully decant the supernatant and add cold 3% glutaraldehyde, being careful not to disturb the pellet. After about an hour of fixation, you should be able to dislodge the pellet and process it as if it were a piece of tissue.
Hope this helps.
Jerry Gagne Senior Biologist Department of Microscopy and Microanalysis Abbott Laboratories (847) 938-5023
We have a student starting to work with human cerebral blood vessels obtained from cadavers.
A literature search has revealed little information on working with cadaver material at the TEM level.
The questions from this student are;
1. Does anyone know of any publications resulting from TEM investigations of samples from human cadavers, in particular cerebral blood vessels and brain / nervous tissue ?
2. The standard EM fixation texts talk about not using formalin to fix for TEM and emphasize speed of primary fixation. My initial results indicate that the ultrastructure of the cadaver cerebral blood vessels has remained unexpectedly intact.
Has anyone else experienced good ultrastructural results in the TEM with cadaver material, embalmed in 10% Neutral Buffered Formalin and after an initial post mortem delay of up to 24 hours ?
- What ultrastructural changes (at the organelle level) did you expect and then what actual ultrastructural changes did you see ?
3. What ultrastructural changes would you expect from such material if it has been in 10% Neutral Buffered Formalin for up to 6 months prior to processing and examination ?
On behalf of our student, thanks for your help.
Allan
-- ------------------------------------------------- Allan Mitchell Technical Manager Otago Electron Microscope Centre C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
----- Original Message ----- } From: {mryder-at-brookes.ac.uk} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, October 17, 2001 8:00 AM
Dear Listers,
We have recently got a free set of cryo-attachment (FC 4) for our old Reichert-Jung ultramicrotome. However, there are two parts missing. One is the alloy steel bridge, which connects the specimen to the ultramicrotome's specimen arm. The other is the steel guide rails, which are used for the mounting of the cryo-chamber. Anyone has these parts or whole cryo-attachment to give away or sell? Please let us know.
Thanks Cheng
----------------------------------------------------- Cheng Huang Australian National University EM Unit, RSBS Box 475, ACT 2601 Canberra, Australia Phone: 61-2- 6125-6553 Fax: 61-2- 6125-3218 http://www.anu.edu.au/EMU/
Is your cooling water cool on return? How long since your dif pump was cleaned and recharged? Vacuum fluid can diminish over time and when its down to half of the initial charge you may have that type of problem. Fluid will work well in some TEMs for years, but suspect the pump if its more than two years since the last fluid change - and less if the pump lived through a couple of mishaps. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, October 18, 2001 5:30 AM, Lilly M. [SMTP:lilly-at-cycy.net] 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, } } We are having a problem with the high vacuum on our HItachi H600 TEM. When } the microscope is } first turned on, the high vacuum is achieved but about 5 to 10 minutes later } it starts } dropping. The problem developed over time. In the beginning, it would drop } by very little and } go back up again. Later on, it started dropping by a lot more and it took } longer to get back } up again. Now, the vacuum is almost competely lost. Does anybody know what } might be causing } this problem? Any tips or suggestions? We appreciate the help and thank you } all in advance.
My colleague Dr. I. Pencea has a proposal for an aplication, if someone is interested in this please contact us.
Best regards, Prof. D. Bojin
" Dear Sir/Madam,
My name is Ion Pencea and I am Assoc. Professor with "Politehnica " of Bucharest, Romania, Materials Science & Engineering Faculty. My research field is materials characterization by WAXD, SAXD and XPS. I have gained a scholarship for training in microscopy investigations (SEM, TEM, AFM, etc.). The training period is of least 6 month and maximum 12 month depending on host agreement. Thus, I am looking for a laboratory which could accept me as a visiting scholar or other position (post doc., etc.). I have some experience in SEM and TEM with a TESLA microscopes (BS 350, BS 540) but I am interested in a modern SEM with EDS / WXD attachments, and of course in TEM and AFM. During this training period I can help with investigation on materials of host interest and other specific work (documentation, papers, reports, etc.). My scholarship will cover my accommodation but if it is possible to get work permission it will be perfect. If you are interested in my application, please contact us.
Most Hitachi microscopes have a seleniod valve that "vents" air into the vacuum line when the SEM/TEM is turned off. This prevents any residual vacuum from sucking oil back into the forlines and causing startup problems.
This seleniod valves get really hot during operation and, over the years, causes the hose to crack. This small crack allows air into the foreline thereby raising the foreline pressure & ultimately the SEM/TEM pressure. As the crack gets larger, the foreline pressure is worse, etc.
The first thing to do is to identify the Selenoid valve(s) in question. Trace the vacuum lines from the rotary pumps to a "T" in the line. The "T" should have a 3/8 in (6mm) OD rubber hose coming from the "T" to the top of the solenoid valve. The solenoid valve will be very hot. Either replace the hose or cut it shorter depending upon it's condition.
Hope this Helps.
Earl Weltmer Scanservice Corp.
----- Original Message ----- } From: "Lilly M." {lilly-at-cycy.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, October 17, 2001 12:30 PM
Judi Bowden writes ...
} ... } Do you see a link between science and art? If so, how and why? } ...
I think you are mixing apples and oranges. "Science" is a process by which we learn ... "art" is something we create and/or observe. However, that is not to say we cannot use a scientific process to learn how to create, or create differently.
We have examined numersous brain samples, both biopsy and autopsy material. The damage you will see due to autolysis will totally obliterate any difference you might have seen due to the use of formalin versus or prior to glutaraledehyde. Length of time storaged in fix will also affect the tissue less that length of time the person was dead before the tissue went into fixative.
The best ultrastructure would be obtained on biopsy material (hard to get), and the next best structure would be from rapid autopsies, like those done in 30 min after Alzheimer's Disease patients have had died for the special studies some folks are doing on rapidly decaying processes.
If one is comparing ultrastructure of one case to another, (s)he needs to note carefully the length of time the person was dead before the tissue went into fix (or the person was embalmed). This time can vary widely.
Sara Miller
On Thu, 18 Oct 2001, Allan Mitchell wrote:
} Date: Thu, 18 Oct 2001 12:07:18 +1300 } From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} } To: microscopy-at-sparc5.microscopy.com } Subject: TEM of cadaver tissue } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Kia Ora all } } We have a student starting to work with human cerebral blood vessels } obtained from cadavers. } } A literature search has revealed little information on working with } cadaver material at the TEM level. } } The questions from this student are; } } 1. Does anyone know of any publications resulting from TEM } investigations of samples from human cadavers, in particular cerebral } blood vessels and brain / nervous tissue ? } } } 2. The standard EM fixation texts talk about not using formalin to } fix for TEM and emphasize speed of primary fixation. My initial } results indicate that the ultrastructure of the cadaver cerebral } blood vessels has remained unexpectedly intact. } } Has anyone else experienced good ultrastructural results in the TEM } with cadaver material, embalmed in 10% Neutral Buffered Formalin and } after an initial post mortem delay of up to 24 hours ? } } } - What ultrastructural changes (at the organelle level) did you } expect and then what actual ultrastructural changes did you see ? } } } 3. What ultrastructural changes would you expect from such material } if it has been in 10% Neutral Buffered Formalin for up to 6 months } prior to processing and examination ? } } On behalf of our student, thanks for your help. } } Allan } } } -- } ------------------------------------------------- } Allan Mitchell } Technical Manager } Otago Electron Microscope Centre } C/-Department of Anatomy and Structural Biology } School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand } } Phone (03) 479 5642 or 479 7301 } Fax (03) 479 7254 } } Unit: http://www.otago.ac.nz/anatomy/emunit/ } Department: http://anatomy.otago.ac.nz/ } } } } "Life is a gift, don't waste it" } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Judy, For most of us, it isn't good enough just to take a picture that shows what we need to see, it also has to look good. I think that is where the art comes in. I've had 3 cover micrographs for the Scanning Journal (www.scanning-fams.org).Most of the images were from my Ph. D. research, but add some color...and you have ART!
Dr.Tina S. Schwach, President Microscopy Consulting Services Inc. 651-681-0112
----- Original Message ----- } From: {"01151938-at-mrc.vic.edu.au"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 16, 2001 6:09 PM
I heard a definition of art when I was in photography school many years ago. I wish I knew the source...
Art is anything, that when abstracted from its surroundings, embibes its viewer with a hieghtened or altered sense of thier own existence.
Soon after is when I went from fine art to scientific imaging because it does this so magically.
If you perform clinical TEM or SEM, I need your input on a College of American Pathology (CAP) reevaluation of the labor time they allot for these procedures. If you're willing to help, please contact me offline, and I will send you a draft of the operating procedures for your comments.
The CAP has CPT codes that determine how much hospital labs can be reimbursed. The descriptions for TEM and SEM procedures are out of date and appear to have been written by a lab director who was not actually an electron microscopist as some of the steps don't make sense. I have been asked by CAP to submit updated lists for consideration for adoption.
We prepare several hundred EM samples per year for Duke hospital, and I have, with the help of my 5 superb techs, listed each step and the time it takes to perform them. If you work in a hospital lab or are contracted to perform EM on human tissue, I would very much appreciate your input. Please contact me.
Thanks in advance, Sara
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Dear Lilly, The two things that I know that can cause that are: 1. the heater on the diffusion pump is not quite heating hot enough to boil the oil, 2. the diffusion pump oil is almost gone. The fact that you achieve high vacuum means that your system is almost working and has no leaks. At 12:30 PM 10/17/01 -0700, you wrote: } } Hi everyone, } } We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is } first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts } dropping. The problem developed over time. In the beginning, it would drop by very little and } go back up again. Later on, it started dropping by a lot more and it took longer to get back } up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing } this problem? Any tips or suggestions? We appreciate the help and thank you all in advance. } } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I have a few opinions about this topic since I am a research microscopist by day and an artist by night. One of my interests involves utilizing elements from science in the production of art. I have an exhibition/performance November 8th--24th at SomArts Gallery in San Francisco that will include some exemplary pieces. You are invited! You can learn more from the web site at http://SaintRubidium.com I will be adding additional descriptions of the art work to the web site before November 1st. Some examples are: a screen made of 6000 Petri dishes; micro art viewed with a light microscope and made with drosophila wings, human blood and copper sulphate; a video of a 30 foot by 30 foot array of data tapes from electrophysiology potassium channel recordings, an eight foot cylinder "totem" made from science journal cover photos; another totem made from radiographic films and transilluminated; a computer controlled 3-slide-projector with stereo sound presentation of botanical scanning electron micrographs made by Douglas Frank in Portland, Oregon.
I find the world of science rich with imagery and fascinating materials that are exciting to use in a manner that brings a different perspective on science to the public while investingating personal issues as well as some issues common to all of humankind and the universe. There are numerous people interested in the intersection of art and science. YLEM just celebrated their 20th anniversary with an excellent exhibit in San Francisco. More information can be found at http://www.ylem.org See their links page for connections to other people and organizations such as the San Francisco Exploratorium. Another resource is the journal Leonardo / the International Society for the Arts,Sciences and Technology. See: http://mitpress2.mit.edu/e-journals/Leonardo/index.html
I hope that these few words explain a little bit of the how and why of the connection between art and science. I must add, however, that I learned early on in my career that there are scientists that find this whole art interest in science repugnant. I'm still not clear why but can only speculate. I also find that the influence of my art on my science is much less clear to me except that it provides some motivation to survive the trudgery that experiments often require. Larry
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The American Chemical Society celebrates National Chemistry Week during November 4-10, 2001. This year's theme is "Celebrating Chemistry and Art." For more information, refer to http://www.acs.org and scroll down to National Chemistry Week.
Kristine Danowski Analytical Sciences Dow Chemical Company Midland, MI 48667 kldanowski-at-dow.com
} Hello: } Could somebody please define for me "home brew and ASR rules"? Does this } phrase refer to how reagents or antibodies are made, in-house vs. } commercially? } Thanks, } Steve Saul } } e-mail: ssaul-at-optronics.com
Dear Tom, I have prepared cell pellets for virus-infected cells many times. If the cells are in 6-well plates, one can process them in the plates thru the dehydration and release the cells (quickly!) with washes of propylene oxide. The cells can then be infiltrated in a P.O. resistent polyproylene test tube and spin down after each change of resin. When embedding, then pipet off the resin to the pellet and tranfer the pellet to beem capsules. Spin them in a piggy-back arrangement with the beem capsule tucked in a microcentrifuge tube. (Cut the microcentrifuge tube so the beem capsule fits) Close the cap, spin and top off with resin. If the cells are in suspension, one must spin after each step of fixation, dehydration, and infiltration before transfering them to beem capsules to embedd. But, you may process suspension cells in the polypropylene tubes which are easy to spin.
Barbara Plowman Univ. of the Pacific School of Dentistry 2155 Webster St. Rm 632 San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu phone: 415-929-6692
I operate an H600 and have seen a similar problem. First, by turned "on" do you mean the high tension voltage (e.g.. 100kv) or do you mean physically starting the system from no pumps running to switching on the pumps and waiting for the DP's to heat up and pump the system down?
If you mean applying the 100kv, I've found that filling the large LN2 trap before switching on the 100kv virtually eliminates any vacuum swings.
The other problem may be that you need to clean the PENNING gauge vacuum sensor. if its heavily coated with junk it will short-out and you see it as vacuum drops and poor levels on the readout. There is a section in the H600 instrument manual that tells you how to clean the gauge. Its very easy to do and takes about 2-3 hours of your time. If needed please contact me off list and I can fax you the pages in the manual.
dz
-----Original Message----- } From: Lilly M. [mailto:lilly-at-cycy.net] Sent: Wednesday, October 17, 2001 3:30 PM To: Microscopy-at-sparc5.microscopy.com
Hi everyone,
We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts dropping. The problem developed over time. In the beginning, it would drop by very little and go back up again. Later on, it started dropping by a lot more and it took longer to get back up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing this problem? Any tips or suggestions? We appreciate the help and thank you all in advance.
} If you are interested in further exploring the connection, I suggest you } contact David Scharf of Los Angeles CA... David is an Artist who produces } color SEM photomicrographs and his work is outstanding.
The reason that I have an SEM is because of David Scharf. In 1976 I bought a copy of his book _Magnifications_ and was entranced by the images. Scharf clearly not only has mastery of the SEM, but also an artist's eye for composition (and a sense of humor, as well). As I looked through the images over and over, I came to believe that the SEM has to be one of the best toys ever invented. It took 25 years, but I now have my own toy.
I believe that science and art support each other very well. One of my other interests is pottery. My scientific background helps me understand which forms will be mechanically strong and which will be fragile, what chemicals in glazes do, how heat in the kiln works, how to conduct meaningful experiments... I feel sorry for those who attempt to create pottery _without_ the benefit of a scientific background. They are at a distinct disadvantage.
Scientific endeavors can also benefit from art. My favorite example is (was) O'Shaughnessy dam on the Scioto river near Columbus Ohio. I say "was" because the original dam no longer exists. The dam was beautiful. It was not just a rectangular block of concrete poured into the path of a river. It had graceful curves and portions of it were terraced into the surrounding rock. Rather than simply plummeting down a abrupt spillway, the water broke over a curved edge, cascaded down the main portion of the breast, and was reintroduced to the river with a reverse curve at the bottom. As a result, the water fell almost silently. More sound came from the small terraced portions, where the water might fall 5 or 10 feet at a time, than from the main spillway. O'Shaughnessy served its purpose well as a dam and was beautiful at the same time.
Hi Judy, There was a great talk this year at the M&M Meeting in Long Beach, CA related to that subject. The MSA Presidential Happenings lecture was presented by Stuart S. Sumida from California State University San Bernardino: From Bones and Stones to Balloons and 'Toons: Bringing Biology and Art Together in Realistically Animated Films. He's a paleontolgist and he has advised Disney animators on animal movement for such films as The Lion King, Mulan, etc. He would be a great person to contact. Perhaps someone has his email address?
On a personal note, I enjoy doing 35mm B&W macro photography. Last year I entered photographs in a juried show. Up to 3 photos could be entered, so I submitted 2 of my favorite 35mm photos and for a laugh I threw in a SEM of pollen grains on a Bumblebee's back. Needless to say I was truly miffed when the SEM was accepted and my others were rejected - ha! Anyway, it made me think about my work and how I approach it. I do try to frame things up in the most esthetically pleasing way and still have it contain all the scientific information that's needed. Even though I've separated my work from my art for many years, I think all microscopists are artists. Is anyone else observing our world so closely? ...well, besides the media;-) good luck with your project, Beth
} Dear Listers } I am presently undetaking an Honours degree at La Trobe University, } Victoria, Australia.I have been researching flower pollen using a Scanning } Electron and Confocal microscopes. } I am particularly interested to hear your views on the following question: } Do you see a link between science and art? If so, how and why? } Thank you for considering my question. } Yours sincerely } Judi Bowden } La Trobe University } Victoria } Australia
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
"Do you see a link between science and art? If so, how and why?"
Judi,
Linking creativity in art and science:
Creativity in art is not substantially different from creativity in science. Creativity in art/science is not a person suddenly struck by a blinding flash of inspiration/insight and then producing a masterpiece. Even the most radical artists/scientists start from classical knowledge and then spend years polishing their technique. [Picasso, Mozart, Einstein] (source: Julio Ottino, lecturer on "Creativity ? an Art View", an accomplished artist and Professor, and Chair, Department of Chemical Engineering, Northwestern University, Evanston, Illinois, USA).
"Even the most talented persons do not reach world class performance in their fields until they have devoted about 10 years or more of rather single minded attention to becoming an expert". What chiefly distinguishes creative thinking from more mundane forms [in art/science] are Ambiguity, Persistence and Knowledge - (i) willingness to accept vaguely defined problem statements and gradually structuring them, (ii) continuing preoccupation with problems over considerable periods of time, and (iii) extensive background knowledge in relevant and potentially relevant areas. (source: H. Simon, Nat'l Acad Sci Proc, 80, 4569-4571, 1983).
Art is the selection from a great number of variables (source: book, Zen and the Art of Motorcycle Maintenance). There are also a great number of variables to select from at the start of a new science project.
Regards,
Ev Osten
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000 U.S.A.
just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
From my background in physics I find this question very intriguing.
Art stands out by its resonance with our experience or by harmony with something within us. We marvel at the expression. Sometimes it expreses a play on patterns with nothing to do with function like designs on ancient potery while other times it's the expression of functional form, the bowmans weapon or the flying gull. We identify, at once the ideal human form in the Greek sculptures and dynamic figures from the caves in France while we see the vision and mastery of the artist.
With science it may not be so different. We paint with a bigger brush with a handle of technology, bristles of training, and paint of intellegent abstractions on a canvas of hypothetical vision. Does not this or that stroke of the brush resemble the observation? If not quite, then improve the stroke, refine the correspondance. Generalize the circle to an ellipse. Now the planetary cycles correspond... The Painting is never done. Maybe it is more like an evolving building with not so many master archetects and many builder craftsmen all who live in the structure they build daily. Their dedicated methodology is required to keep the structure sound with each new change.
But science is an art. It is an art of knowing, a craft of many participants. Mere scientific method cannot make breakthrough discoveries. It is an art of creative nurturing of insight correctly applied to each step.
The same honesty requred in the scientific method may also be required by any artist to acheive her mastery of technique.. The same aesthetic experience of the artist and viewer may also be part of the experience of scientist and student.
My $00.04 worth,
Dave -- David Barnard Wadsworth Ctr NYS Dept Health Albany NY 12201-0509 barnard-at-wadsworth.org 518 473-5299 voice 518 474-7992 fax
I am trying to do some SEM on nematodes with elaborate structures on their front ends. Unfortunately, these critters secrete some sort of sticky residue around these structures that makes it difficult to get good, clean specimens. The "stuff"is likely some sort of glycoprotein goo and I should (hopefully) be able to rinse it off with something, without damaging the integrity of the collagenous cuticle and without giving me other sorts of artifacts. Any ideas? -- ______________________________________________ Dan Bumbarger Graduate Student Depts. of Biology and Nematology University of California, Riverside Riverside, CA 92521 ______________________________________________
"With a little more deliberation in choice of pursuits, all men would perhaps become essentially students and observers, for certainly their nature and destiny are interesting to all alike. In accumulating property for ourselves or our posterity, in founding a family or a state, or acquiring fame even, we are mortal; but in dealing with truth we are immortal, and need fear no change nor accident."
Dear Michelle, Had the same problem when I was starting out. It turned out that I was alternating with someone who was using tungsten film and removed the daylight filter for his photography. On another occasion when I was the lone user, I found that there were lots of voltage changes during the day and photos taken at one time were different from those taken at another. The immediate way to handle the voltage problem is to check the power supply voltage level for every exposure. When I had a class taking photos, I used a tabular sign-in sheet with columns for: Power supply setting; Iris diaphragm open; field diaphragm adjusted; objective; time of photo; trinocular prism setting (1,2, or 3); etc. I think you get the point. If your line voltage is at fault, the only way you can prove it is to have someone perform a time study to record those changes over a 24hr period at your outlet. The next problem that I have seen is caused by picking a roll of film on Monday that is designated for use with a tungsten source and another roll on Thursday that is designated for use with daylight. Oh, yes! AND FINALLY, if your photo detector is set for spot metering rather than averaging, a small change in location can/will have a great effect on the result, which will appear to be unpredictable. From all of the above, your best bet is the proper compensating filter when using artificial light (LBD (Olympus) for daylight-type(5500K) and LBT for tungsten-type(3400K) films) For years I have treasured a book I acquired from Olympus entitled, "How To Improve Photography Through the Microscope" (Part # M132E-0886T). There is another that I acquired from Leitz before it became Leica.
Hope that one small part of this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: mryder-at-brookes.ac.uk } Sent: Wednesday, October 17, 2001 8:00 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist:LM color changes in photographs } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (mryder-at-brookes.ac.uk) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } October 17, 2001 at 05:31:30 } -------------------------------------------------------------------------- } - } } Email: mryder-at-brookes.ac.uk } Name: Michelle Ryder } } Organization: Oxford Brookes University } } Education: Graduate College } } Location: Headington Campus, Gipsy Lane, Oxford, OX3 0BP } } Question: Why do my wholemount lowish power root } photographs sometimes come out yellow, on the } same film, with the same lighting. Sometimes } the same root taken a few minutes later will } be yellow when it was the usual white on a previous frame. } } } } -------------------------------------------------------------------------- } - } }
"This method did not work in our hands." "I've been looking at histologic sections for 20 years, and I never made that connection." "How did you get these preparations to have such low background?" If there is no art in the application of the principles of science to the business of discovery, then there is no discovery.
Here's to the art(full) scientist who discovers what others have missed, because she sees the truth in corn while everyone else is treating it like food.
Regards to all,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
Lilly, I've seen this kind of behavior if the oil level in the diffusion pump gets low (I'm assuming that it is diffusion pumped). What happens is that the oil starts to boil and the DP pumps. When the oil is all boiled off, it stops pumping until some condensed oil runs down the side and back into the boiler section where it can boil and pump a little more.
The solution is to remove the DP, clean it and recharge it with the proper amount of the proper oil. Generally this problem occurs when you either have a large air leak or the DP is frequently vented when still quite hot, a definite no-no.
Ken Converse owner Quality Images third party SEM service Delta, PA
Lilly M. 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, } } We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is } first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts } dropping. The problem developed over time. In the beginning, it would drop by very little and } go back up again. Later on, it started dropping by a lot more and it took longer to get back } up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing } this problem? Any tips or suggestions? We appreciate the help and thank you all in advance. } } }
Shaf, My wife will tell you art can be taught just like science. There are basic principles involved that anyone can learn. Just like science, though, some have more "innate", "natural",( you choose the term) ability. This is why there are some outstanding artists and some outstanding scientists. The creativity is the difference between the norm and the great in both cases. The same holds in most every field of endeavor that I can think of.
Ken Converse
michael shaffer 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 } -----------------------------------------------------------------------. } } } Judi Bowden writes ... } } } ... } } Do you see a link between science and art? If so, how and why? } } ... } } } I think you are mixing apples and oranges. "Science" is a process by } which we learn ... "art" is something we create and/or observe. However, } that is not to say we cannot use a scientific process to learn how to } create, or create differently. } } my 0.02 ... shAf :o) } } } }
I've looked at numerous post mortem retina specimens. I agree with Sara, though do find that glut fixation is better than form, even though there are P/M effects to consider. Unfortunately it's not always possible to say that a tissue from, say, 8 hours P/M will be better than one from, say 12 hours. Other variables, such as temperature of the body during that time come into play. Sometimes 8 (or 12 or even 15) hours P/M is fine, sometimes useless. Obviously however, less is generally better. Some cells seem a lot more "fragile" than others and deteriorate very rapidly - it's useful to find, for your tissue, which these are and use them as a rough measure of P/M effects. Blood vessels do seem pretty tough and I've seen quite good vessels when surrounding tissue is a mess. I haven't had to embed much for EM after long term storage so can't answer question 3, but have done immuno work on form fixed material up to years old and it was fine for LM. As for what U/S changes to expect in P/M material, that's a question that could take up a whole book! Just look carefully and compare your own specimens with each other and with what you know to be good U/S. It's all fairly obvious with experience.
Diana
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Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Yes it is possible to check electronic devices. From our experience it may give better results if you are applying a load (mechanical and/or thermal) to the device or much better to the entire pcb. Sometimes these tiny cracks are occurring only if the load is applied.
email: mklein-at-visitec-em.de WWW: http://www.visitec-em.de * Home of world's largest SEM *
-----Ursprüngliche Nachricht----- Von: Mark Riggs [mailto:Mark.Riggs-at-asml.com] Gesendet: Donnerstag, 18. Oktober 2001 22:11 An: microscopy-at-sparc5.microscopy.com Betreff: semming a capacitor
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just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
} } } } "Do you see a link between science and art? If so, how and why?" } } Judi, } } Linking creativity in art and science: } } Creativity in art is not substantially different from creativity in } science. Creativity in art/science is not a person suddenly struck by a } blinding flash of inspiration/insight and then producing a masterpiece. } Even the most radical artists/scientists start from classical knowledge and } then spend years polishing their technique. [Picasso, Mozart, Einstein] } (source: Julio Ottino, lecturer on "Creativity ? an Art View", an } accomplished artist and Professor, and Chair, Department of Chemical } Engineering, Northwestern University, Evanston, Illinois, USA). } } "Even the most talented persons do not reach world class performance in } their fields until they have devoted about 10 years or more of rather } single minded attention to becoming an expert". What chiefly distinguishes } creative thinking from more mundane forms [in art/science] are Ambiguity, } Persistence and Knowledge - (i) willingness to accept vaguely defined } problem statements and gradually structuring them, (ii) continuing } preoccupation with problems over considerable periods of time, and (iii) } extensive background knowledge in relevant and potentially relevant areas. } (source: H. Simon, Nat'l Acad Sci Proc, 80, 4569-4571, 1983). } } Art is the selection from a great number of variables (source: book, Zen } and the Art of Motorcycle Maintenance). There are also a great number of } variables to select from at the start of a new science project. } } Creativity is not bounded by art or science. The closer you get to the leading edge the more science becomes an art. The only difference is in art you have critics and in science you face peer review.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
Hi Dan, We used to remove the stylet secretions from rot-knot and soybean cyst juveniles (very sticky stuff) by adding 1 part of a high pH buffer (0.1 M Tris-NaOH, somewhere around pH 11-12) for 2 minutes or so. Of course, we were attempting to collect the secretions and not preserve the nematode. Although they seemed to be fine, as we could usually do this for several days after washing the 'tode. John Shields EM Lab University of Georgia
On 18 Oct 2001, at 14:54, Dan Bumbarger wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I am trying to do some SEM on nematodes with elaborate structures on } their front ends. Unfortunately, these critters secrete some sort of } sticky residue around these structures that makes it difficult to get } good, clean specimens. The "stuff"is likely some sort of glycoprotein } goo and I should (hopefully) be able to rinse it off with something, } without damaging the integrity of the collagenous cuticle and without } giving me other sorts of artifacts. Any ideas? -- } ______________________________________________ Dan Bumbarger Graduate } Student Depts. of Biology and Nematology University of California, } Riverside Riverside, CA 92521 } ______________________________________________ } } } "With a little more deliberation in choice of pursuits, all men would } perhaps become essentially students and observers, for certainly their } nature and destiny are interesting to all alike. In accumulating } property for ourselves or our posterity, in } founding a family or a state, or acquiring fame even, we are } mortal; but in dealing with truth we are immortal, and need fear no } change nor accident." } } } -H.D. Thoreau, "Walden" }
Well, yes and no. I agree that creativity is basal to both art and science, but surely the two are fundamentally distinct at least as far as their aims are concerned. The aim of science is to discover the truth (whatever that may be! - see e.g. Popper) about the natural world and its properties. Scientists use their creative imagination to devise ways of discovering facts about the world. Artists use their creative imagination to create objects -artefacts- which never have been of this world before. Scientists also create artefacts, but usually blush when one is pointed out to them. I find it fascinating that the most imaginative scientists are also profoundly aware of the arts, and are frequently very accomplished artists. Equally many artists are intrigued by the findings of science, if not actually by the methods, so the possibility exists of considerable common ground between the two activities as far as the mental / intellectual processes are concerned. But the key features that distinguish science from art are the accumulation and evaluation of information, the use quantitative analysis and of logic, and the application of a rigorous scientific methodology to distinguish fact from non-fact. None of these things are fundamental to art, indeed with a few exceptions their use seems to be at best optional, if not scorned altogether. In other words it is the prerogative of the artist to let the mind be totally free of the contraints of reality. Scientists do not have that luxury.
Chris
} Creativity is not bounded by art or science. The closer you get to the } leading edge the more science becomes an art. The only difference is in art } you have critics and in science you face peer review. } } Gordon } } Gordon Couger } Stillwater, OK } www.couger.com/gcouger } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
when was the last time your DP oil was replaced? It could be 'cracked' or, as was explained to me by a JEOL engineer, saturated by trying to remove more air from the column than it was capable due to a variety of reasons like too short a prepump(roughing) time, and/or improper valving sequences from rough pumping to high vac or a small column leak.
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Hi everyone,
We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts dropping. The problem developed over time. In the beginning, it would drop by very little and go back up again. Later on, it started dropping by a lot more and it took longer to get back up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing this problem? Any tips or suggestions? We appreciate the help and thank you all in advance.
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Hello semmers, Since I have had such great help in the past from all of you, I have decided to once again seek information from this great source. I am wondering if any of you use reactive ion etching, or even more helpful would ICP-RIE. If so, it would be great if you could answer some questions for me. Please contact me off line, as it isn't really a microscopy question. Thanks again Nick
Nicol Aitken Sample Preparation and Imaging Specialist Research and Development Semiconductor Insights Inc. email:nicol-at-semiconductor.com (613)599-6500 ext.4300
a week ago I wrote asking about a problem related to image distortion on our SEM. Thanks to all of you who wrote suggestions to us. the technicians are studying the solution of the problem.
My question now is related to sample preparation of paper for studying the thickness of the fibers. The idea is to obtain cross sections of paper sheets showing the sections of the fibers. How would you prepare it in order to avoid swelling or deformation of the sections? Embedding would not be the ideal method since the surface of the same paper has to be observed.
Any suggestions will be very much appreciated.
Thanks a lot
Silvia Montoro CERIDE G|emes 3450 3000 Santa Fe - Argentina csedax-at-ceride.gov.ar
Mark: a lot depends on what type of capacitor it is. More detail about the cap would be helpful. In my experience, small ceramic surface-mount caps are more prone to problems in the area than large metal-can electrolytics. I agree with Martin Klein that examining the cap in its native habitat would be the best, but it all depends on how big your SEM chamber is and where you suspect the cracks might be occurring. If they are between the cap and the board; they'll be really hard to see in the SEM. A word of advice: this is best done in a field-emission SEM or variable-pressure SEM because charging is going to be a big problem and I'm assuming you don't want to coat the board with gold or carbon.
Mark Riggs 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 } -----------------------------------------------------------------------. } } just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) SC Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
We have a Polaron Turbo Coater carbon evaporator, model E6700, probably 10 - 15 years old, which is experiencing some problems. In spite of acceptable vacuum and functioning pirani gauges, the penning gauge is not turning on, and one of the fuses is blowing every time we turn it on.
We don't have schematics for any of the boards in the instrument, just a schematic which gives an overview of the entire instrument, but no specifics of individual boards.
If anyone out there has schematics for any boards in this instrument, and would be willing to share this info, could you please contact me off line?
Thanks, Wharton Sinkler UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017
Hello, I'm trying to find a couple of recommended references for phytoplankton and other small items to be fixed/dried for SEM. The samples have the problem of getting lost during the CPD process. I have the following, but I cannot find the book Scanning Electron Microscopy II. Has anyone seen these references and are they correct as I have them:?
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.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Dear Judy, To your question: Do you see a link between science and art? If so, how and why? The mediating event between the interiority and exteriority of our experience of the world is in the aesthetic moment according to Jamesian philosophy. It forges the union between past and future, between certitude and expectation. As many other have said previously in this discussion -especially David Barnard's excellent observation - their work often hinges on the awareness of beauty within the work. It ties together the craft of rigorous method with optimism of our theory.
I have spent thirty years working in photography in all its aspects, from street documentary to fashion studio work, from mug shots to gallery work. One of the most exciting bodies of work that I ever produced was as student of EM at the VA Center in Albany, NY, now sadly closed. I shot something like 4 times the amount of film than most students did. Why? Because the electron microscope, especially the TEM, is nothing more than a camera, but what a wonderful camera it is! It investigates a landscape, searches for a recognizable form, establishes proportions and makes measurements, always looking for 'significance' however that may be defined . In short, exactly the same thing I try to do in conventional landscape photography. It' s all a part of the attempt to make sense of the perceptible world, or to make the imperceptible visible and then intelligible.
I recommend to you the following: Art Forms in Nature, Ernst Haeckel and Karl Blossfeldt's Art Forms in the Plant World
Fred and Michelle, I'd like to add a few additional and perhaps unrelated points to getting consistent results. Having done a a lot! of photomicrography especially for publication in text books, another method for getting consistent results is to buy a block of film that is all from the same lot (check the labels). I also learned that film is made to have a known storage or shelf time until it reaches the optimum color balance, so you have to find out what that is as well. Finally, I had to find a processor that ran test strips at the beginning of the day to assure that the process was correct, then I made sure that they did my film first thing in the morning. Regarding filters, I had to buy a set of light balancing and color compensating (CC) filters to get "perfect" color balance. Naturally, I always bracketed by about 1 stop in 1/3 stop increments as my time was more expensive than the film. I learned that the voltage fluctuated slightly (dropped) and had to turn the lamp on and wait about 30 min to get it stable. Obviously I was doing very critical color work and this all may be overkill for you. Regards from an oldtimer whose using digital now :-) Damian } } Dear Michelle, } Had the same problem when I was starting out. It turned out that I } was alternating with someone who was using tungsten film and removed the } daylight filter for his photography. On another occasion when I was the } lone user, I found that there were lots of voltage changes during the day } and photos taken at one time were different from those taken at another. } The immediate way to handle the voltage problem is to check the power supply } voltage level for every exposure. } When I had a class taking photos, I used a tabular sign-in sheet } with columns for: Power supply setting; Iris diaphragm open; field } diaphragm adjusted; objective; time of photo; trinocular prism setting (1,2, } or 3); etc. I think you get the point. If your line voltage is at fault, } the only way you can prove it is to have someone perform a time study to } record those changes over a 24hr period at your outlet. } The next problem that I have seen is caused by picking a roll of } film on Monday that is designated for use with a tungsten source and another } roll on Thursday that is designated for use with daylight. } Oh, yes! AND FINALLY, if your photo detector is set for spot } metering rather than averaging, a small change in location can/will have a } great effect on the result, which will appear to be unpredictable. } From all of the above, your best bet is the proper compensating } filter when using artificial light (LBD (Olympus) for daylight-type(5500K) } and LBT for tungsten-type(3400K) films) } For years I have treasured a book I acquired from Olympus entitled, } "How To Improve Photography Through the Microscope" (Part # M132E-0886T). } There is another that I acquired from Leitz before it became Leica. } } Hope that one small part of this helps, } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } } } } ---------- } } From: mryder-at-brookes.ac.uk } } Sent: Wednesday, October 17, 2001 8:00 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Ask-A-Microscopist:LM color changes in photographs } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (mryder-at-brookes.ac.uk) from } } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } } October 17, 2001 at 05:31:30 } } -------------------------------------------------------------------------- } } - } } } } Email: mryder-at-brookes.ac.uk } } Name: Michelle Ryder } } } } Organization: Oxford Brookes University } } } } Education: Graduate College } } } } Location: Headington Campus, Gipsy Lane, Oxford, OX3 0BP } } } } Question: Why do my wholemount lowish power root } } photographs sometimes come out yellow, on the } } same film, with the same lighting. Sometimes } } the same root taken a few minutes later will } } be yellow when it was the usual white on a previous frame. } } } } } } } } -------------------------------------------------------------------------- } } - } } } } } }
Silvia Montoro wrote: ========================================================= ............... My question now is related to sample preparation of paper for studying the thickness of the fibers. The idea is to obtain cross sections of paper sheets showing the sections of the fibers. How would you prepare it in order to avoid swelling or deformation of the sections? Embedding would not be the ideal method since the surface of the same paper has to be observed.
Any suggestions will be very much appreciated. ========================================================== You are quite correct to be concerned about possible swelling and/or deformation of the paper structure if embedded. However, another concern is the possible swelling of the fibers themselves with the embedding resin, thereby giving you a diameter figure that is much larger than reality.
Our approach has been to gold coat both sides in a sputter coater before embedding. It is always a trial and error exercise to determine the "right" thickness of the gold layer, the purpose for which is to a) passivate individual fibers to reduce the tendency of the embedding resin to penetrate and swell individual fibers and b) reduce the "elasticity", or the ability of the larger structure to be changed by the embedding media. We find that Pt is somewhat better than gold because of its smaller grain size and the greater hardness results in a lowered tendency for the metal to "smear".
We have always had our best results with our own SPI-Pon™ 812 Epoxy Embedding Resin Kit but we believe some of the other Epon® "substitutes" would work just as well.
Using room temperature sectioning, you can then thin section the block and by TEM, the gold layer acts as a nice decoration line outlining the fibers. If there has been excessive fiber swelling, usually there are signs that this is happening, at least once one develops experience in looking at such micrographs. If you don't have a TEM, you can look by SEM at the "faced-off -piece", that is, the block from which the last section was taken. So long as you are using a thin layer of conductive metal, usually there is enough contrast from the gold decoration line to resolve well the individual fibers . Obviously this technique will work better with thinner than it will with thicker papers. It might not be the perfect solution but it does permit one to make measurements on the fiber diameters. Also, at times a short plasma etching with oxygen on the faced-off-surface will enhance further the contrast between the gold layer and the fibers and embedding resin.
If we would be doing this kind of work today, and wanting to look at the faced-off-piece by SEM, instead of coating with gold we would coat with 1 nm of osmium metal in the OPC Osmium Coater. Although a heavy metal, that thickness is thin enough that it does not interfere with being able to see contrast from the underlying gold (decoration) layer.
Note: You should be using a "materials science" rather than a life science (LS) diamond knife since they are cheaper than an LS knife and also, the inorganics in most papers would "kill" a LS knife for future use on LS samples.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
on 10/18/01 11:32 PM, Robert Underwood at underwoo-at-u.washington.edu wrote:
} } I heard a definition of art when I was in photography school many years } ago. I wish I knew the source... } } Art is anything, that when abstracted from its surroundings, embibes its } viewer with a hieghtened or altered sense of thier own existence. } Dear Robert and other listers, When I was an undergraduate, I was arrogant enough to propose a definition of art--one which would include such pieces as a number of bricks arranged in a neat pile (on display at MOMA). I defined art as the projection of the universe onto some chosen set of limitations. Good art enlightens the artist and audience about the nature of the universe by reducing it to an understandable piece; whereas bad art is unsuccessful in this enlightenment (so the value judgement rests with the audience). Not only do I still think that this is a valid definition, but I can also see how this applies very obviously to science. IMHO, therefore, science and art are not separate, but science is art. Yours, Bill Tivol
I'm not sure that this is off topic. Of course, Nestor can correct me if I am wrong.
I do RIE and ICP-RIE etching of IC passivation followed by SEM imaging on a somewhat routine basis. ICP is a new method and is quite improved over traditional RIE methods. Passivation removal is quite difficult based on power, pressure and gasses. For oxide, I use CF4 and O2 at a pressure of about 10mT. I'm not certain of the power.
My particular work in this area is to SEM image ICs which have been de-passivated. RIE and ICP-RIE accomplish this in different final outcomes. I find dramatic differences between packaged die and bare die.
gary g.
At 09:39 AM 10/19/2001, you wrote:
} Hello semmers, } Since I have had such great help in the past from all of you, I have } decided to once again seek information from this great source. I am } wondering if any of you use reactive ion etching, or even more helpful would } ICP-RIE. If so, it would be great if you could answer some questions for me. } Please contact me off line, as it isn't really a microscopy question. } Thanks again } Nick } } } } Nicol Aitken } Sample Preparation and Imaging Specialist } Research and Development } Semiconductor Insights Inc. } email:nicol-at-semiconductor.com } (613)599-6500 ext.4300
Dear All: Thank you for those who replied to my posting regarding anti-amylase antibody search. I eventually located one at the Chemicon. Now, I am looking for a polyclonal anti-synaptophysin antibody. I know that there are many companies carry this product. But I would like to know users' experiences before I decide on which one to purchase from. If you have used this antibody from a particular company, would you share the following information with me. Thank you a million in advance.
Supplier name: Samples labeled: (cell culture or tissue type, species...) Labeling type: (immuno fluorescence, immunoperoxidase, or immunogold?) Labeling procedure: (pre-embedding or post-embedding?) Fixative used: Embedding method used:
By the way, for those who do not have Linscott's directory, Try www.sciquest.com. I do my antibody search on this site. It is very easy to use.
The type of capacitor determines what you have to do. Titanate type chip capacitors consist of alternating metal electrode sheets sandwiched between the dielectric layers, both of which are laid down by printing methods and then fired into a monolithic structure. End metallization is applied so that the alternate protruding plate layers are electrically joined to a metal coating on each end. It is a robust structure but because it is made of very hard, rigid material, locating an internal fracture can be difficult. A fracture that extends to the outside can usually be seen in the SEM without coating the specimen. The tendency for dielectrics like titanates to charge is highly dependent on the orientation of the subject to the beam. One can usually tilt the faces of the capacitor to find an angle where charging is not a problem and then simply search for the crack. Using the backscatter detector instead of the secondary electron detector will also help to minimize charging interference.
Apart from the SEM, a fluorescent dye can also be used for an exterior crack. Magniflux makes such compositions but you can find others by doing a search with the terms "dye penetrant" and "non-destructive testing". The usual procedure is to place the cap in the dye inside a small vessel in which the pressure can be reduced. The vessel is partially evacuated so that upon return to room pressure the dye is drawn into the crack. The cap is then quickly rinsed and observed under a ultraviolet light with a low power microscope. Dye tends to creep out of the crack revealing its position.
Internal cracks can take different forms. One problem type is where the end metallization separates from the edge of one or more of the plates. This is a common source of temperature dependent, intermittent faults. In this case some destructive sample prep is required and there is a chance that the defect will be lost or "healed" in the course of the prep. One can determine the edge direction of the plates and then, using metallographic methods, grind and polish off the exterior ceramic coating to expose the edges of the plates. Don't go beyond the minimum necessary to expose the edges. Use a nap-less cloth for polishing, very light pressure and diamond abrasive to minimize smearing of the metallization. The point where each plate intersects it's respective end cap is a critical area to check for discontinuities. In the SEM you can now use the secondary detector and some charging to your advantage by grounding one endcap and leaving the other electrically floating. A plate that is charging up while its companions are not reveals where a discontinuity exists. Conversely, a plate that fails to charge when its companions on the same endcap are doing so reveals a short to the other endcap. Simple cracks in the internal dielectric can also be revealed by orienting the subject to where charge accumulates and edge effects accentuate the charging. If your SEM is set up for specimen current imaging, compare what you see in this mode with the results of SEI and BEI.
Good luck.
John Twilley Conservation Scientist
Mark Riggs wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
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Morning Dan, One might suggest warm physiological saline, but some will recommend you try what one of my children called 'goozyme', three others called 'spit', or I call, saliva, but most will recommend amylase, or some other -ase, which will cost unless you find some in a nearby refrigerator. While there are some who will interrupt this conversation by mentioning that the former -ase (that is, amyl-), only hydrolyzes glocogen, I have seen it do nicely on what used to be called mucous secretions, if your source of amylase is saliva (p.c.?). NOTE1: All protein-goo is preserved (cross-linked) by fixation longer than necessary to terminate (p.c.?) the subject (p.c.?). If it must be removed, then one should try to do it prior to fixation - not necessarily prior to termination (p.c.?). NOTE2: The above methods can be tried in any sequence for good or ill, but be forewarned that your actions will be misunderstood if you are seen by colleagues to be expectorating (p.c.?) at a worm. NOTE3: Of course, if you continue to work with worms with rhinitis, you must suffer the consequences. In all seriousness, however, if one's worm has a cold, could one preempt the goo with oxymetazoline hydrochloride. This problem, of course, will remind us all that one cannot grasp a live hagfish [old or young!], because it ties itself in a knot, secretes a bucket of goo, and then extricates itself by moving the knot (a posteriori), all of which hasn't much to do with your goo Dan, but does present a much more messy problem that others [I know not who!] have had, which should make you feel better about your, only slightly, goo'd animal. It is, after all, Monday morning, and having seen our grandson off on Saturday, Grandma and Grumpy spent the rest of the weekend, unsuccessfully trying to catch up on sleep. As always, we hope that if the above doesn't bring immediate ease of the problem, it will, at least engender a smile.
Regards and commiseration to all those who have wormy problems,
Fred Monson
(p.c.) = (polite conversation?), of course!
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) So, I will answer the phone with my name, not the name of my workplace. West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Dan Bumbarger } Sent: Thursday, October 18, 2001 5:54 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Dirty Nematodes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying to do some SEM on nematodes with elaborate structures on } their front ends. Unfortunately, these critters secrete some sort of } sticky residue around these structures that makes it difficult to get } good, clean specimens. The "stuff"is likely some sort of glycoprotein } goo and I should (hopefully) be able to rinse it off with something, } without damaging the integrity of the collagenous cuticle and without } giving me other sorts of artifacts. Any ideas? } -- } ______________________________________________ } Dan Bumbarger } Graduate Student } Depts. of Biology and Nematology } University of California, Riverside } Riverside, CA 92521 } ______________________________________________ } } } "With a little more deliberation in choice of pursuits, all men would } perhaps become essentially students and observers, for certainly } their nature and destiny are interesting to all alike. In } accumulating property for ourselves or our posterity, in } founding a family or a state, or acquiring fame even, we are } mortal; but in dealing with truth we are immortal, and need fear no } change nor accident." } } } -H.D. Thoreau, "Walden" } } }
I realize this topic has been hammered out in this forum a few times (for everything except the Nikon Optiphot superwide trinocular) ......can someone direct me to any information that would be helpful? ("I can do this .......yeah!!!") Thanks,
Hello. I am a marine biology graduate student at Memorial University of Newfoundland, Canada. One of my research topic is to study the microstructure of the statolith in the pelagic marine tunicate (Larvacea), Oikopleura vanhoeffeni. The statolith is a small cylindrical calcium build up (10 micrometer diameter, 20 micrometer height), attached to the brain of the tunicate. This bony structure is contained in the membraneous statocyst and probably works as a gravity detector. I would like to isolate this structure for SEM and TEM analyses. After this, Xray microcroprobe and other procedures will follow.
Dissecting the brain is easy but taking out the statolith from the statocyst is the difficult part. Visualizing the statolith can be done by calcium specific stain, alizarin red. I have tried to dissect the statolith out by using fine insect pins ( { 20 micron tip size), but the pins are too large. Doing this under dissecting scope is impossible since 4x magnification does not allow me to do such microdissection.
I tried to dissolve the statocyst with bleach and hydrogen peroxide to isolate inorganic statolith. However, after this procedure, I can no longer visualize the statolith since alizarin red stain dissolves instantaneously in these chemicals.
Sonication was also done to rupture the statocyst and pop the statolith out but it did not work.
Do you have any suggestion on how I can separate this structure? Sincerely, Nami Choe ____________________________________________________________________________
Nami Choe Ocean Science Centre Memorial University of Newfoundland St. John's, NF A1C 5S7 Canada phone:(709)737-3247 fax:(709)737-3220
I don't know anything about the structure you describe, but check out these references:
Howell DN, Miller SE. Identification of viral infection by confocal microscopy. 1999. Methods in Enzymology. Academic Press, 307:573-91.
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.
Briefly, we make "vibratome" sections, stain the slices in propidium iodide, and examine the tissue by confocal microscopy for areas of interest. We also sometimes use antibodies with a fluorescent tag. We then cut out a smaller area including the structure of interest (1 x 1 mm) in a shape of the state of Nevada so that you will know if the section gets flipped over.
If your structure is something that you can recognize morphologically, and you can deal with a 100 to 400 um thick section, this might work.
Good luck,
Sara Miller
On Mon, 22 Oct 2001, Nami Choe wrote:
} Date: Mon, 22 Oct 2001 15:37:43 -0230 (NDT) } From: Nami Choe {c56nc-at-morgan.ucs.mun.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: microdissection before SEM/TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopy members; } } Hello. I am a marine biology graduate student at Memorial University of } Newfoundland, Canada. One of my research topic is to study the } microstructure of the statolith in the pelagic marine tunicate (Larvacea), } Oikopleura vanhoeffeni. The statolith is a small cylindrical calcium } build up (10 micrometer diameter, 20 micrometer height), attached to the } brain of the tunicate. This bony structure is contained in the } membraneous statocyst and probably works as a gravity detector. } I would like to isolate this structure for SEM and TEM analyses. After } this, Xray microcroprobe and other procedures will follow. } } Dissecting the brain is easy but taking out the statolith from the } statocyst is the difficult part. Visualizing the statolith can be done by } calcium specific stain, alizarin red. I have tried to dissect the } statolith out by using fine insect pins ( { 20 micron tip size), but the } pins are too large. Doing this under dissecting scope is impossible since } 4x magnification does not allow me to do such microdissection. } } I tried to dissolve the statocyst with bleach and hydrogen peroxide to } isolate inorganic statolith. However, after this procedure, I can no } longer visualize the statolith since alizarin red stain dissolves } instantaneously in these chemicals. } } Sonication was also done to rupture the statocyst and pop the statolith } out but it did not work. } } Do you have any suggestion on how I can separate this structure? } Sincerely, } Nami Choe } ____________________________________________________________________________ } } Nami Choe } Ocean Science Centre } Memorial University of Newfoundland } St. John's, NF } A1C 5S7 Canada } phone:(709)737-3247 } fax:(709)737-3220 } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
From root Mon Oct 22 17:12:26 2001 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA28070 for dist-Microscopy; Mon, 22 Oct 2001 16:34:28 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA28067 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 22 Oct 2001 16:33:57 -0500 (CDT) Received: from renko.ucs.ed.ac.uk (renko.ucs.ed.ac.uk [129.215.13.3]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA28060 for {microscopy-at-sparc5.microscopy.com} ; Mon, 22 Oct 2001 16:33:46 -0500 (CDT) Received: from MESH900 (dialup-169.publab.ed.ac.uk [129.215.38.169]) by renko.ucs.ed.ac.uk (8.8.7/8.8.7) with SMTP id WAA02642; Mon, 22 Oct 2001 22:30:18 +0100 (BST) Message-ID: {000d01c15b41$4cf1c740$a926d781-at-ed.ac.uk} Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk}
Nami My guess is you are probably already on the right tack with your attempts to isolate the inorganic statoliths by treatment with hypochlorite and peroxide. If you next wash these reagents out of your preparation, Alizarin red will again stain the statoliths, so you can see them. The main problem may be to handle the statoliths during these treatments. One effective way of containing such small objects through a series of reactions or fixation/dehydration steps is to contain them in a Millepore filter unit, exchanging the solutions using a disposable syringe. There is a wide range of filter media with different chemical compositions and pore sizes to choose from. The majority will retain object in the size range of your statoliths.
hope this helps Chris Jeffree
----- Original Message ----- } From: "Nami Choe" {c56nc-at-morgan.ucs.mun.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, October 22, 2001 7:07 PM
to all, My question concerns connecting a light source to a metallograph. I have inherited a Zeiss metallograph with a mismatched light source, Optiquip. The unit arrived without a collar connecting the lamp to the metallograph. The equipment is consists of Zeiss metallograph (gray), 47 17 65 Optiquip light source (creme), model 770. I would appreciate comments from anyone having experience connecting these 2 components or knows of a supplier that can assist. Apparently, the length of the collar is critical and perhaps the diameter.
We have mounting solutions for the Coolpix cameras to fit most microscopes through a C-mount, phototube or eyepiece tube. We have the cameras and accessories as well. A number of configurations are available depending on the microscope. I would be happy to assist in finding the proper mounting, for most instruments we need brand and model number to determine requirements.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
To: Great Minds in the Halls of Science Wisdom!
I realize this topic has been hammered out in this forum a few times (for everything except the Nikon Optiphot superwide trinocular) ......can someone direct me to any information that would be helpful? ("I can do this .......yeah!!!") Thanks,
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (charlted-at-mcmaster.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, October 22, 2001 at 12:59:02 ---------------------------------------------------------------------------
Question: I am wanting to find out better embedding techniques for light microscopy which will allow me to remove a lot of the chlorophyll present in the plant tissue but retain as much anthocyanin as possible. I am currently trying to determine anthocyanin location in a particular plant leaf. My first embedding process involved using a 5% gluteraldehyde and phosphate buffer, followed by a dehydration in increasing concentrations of ethanol and then embedding using JB-4 epoxy resin kit. However, no anthocyanin is present after the above process. Do you have any ideas of how I might retain the anthocyanin, reduce the chlorophyll and observe the pigment location using a light microscope. EM will be used at a later date, but for now I am curious on how do do the above process. Thanks for your help, Emily Charlton, McMaster University, CANADA
The Department of Biological Sciences at SUNY-Brockport is searching for a Chairman. The vacancy announcement follows. We have an Hitachi 7000, several ulytramicrotomes including an Ultracut S, digital darkroom and wet darkroom facilities, along with several excellent LM with digital imaging systems.
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The Department of Biological Sciences invites applications for a chair and professor. Foremost, the Department is seeking an energetic, visionary chair who will develop and enhance a balanced undergraduate major and Master’s degree program. Required: Ph.D. in biology or related field; strong teaching, scholarship, grant activity and administrative skills; ability to work collegially in culturally diverse environment. The disciplinary expertise of the candidates is a secondary consideration. In August 2001 the Department moved into a newly renovated building ($12 million) with state-of-the-art teaching and research facilities. Currently, the Department has strengths in environmental sciences, cell/molecular biology, and vertebrate biology.
Submit letter of application, resume, transcript showing highest earned degree, three letters of reference, and brief statements of leadership philosophy, teaching philosophy, research plans and equipment requirements.
Salary is competitive. Starting date is August 2002. Beginning review date for applications is December 15, 2001. Applications will be accepted until January 15, 2002 or until the position is filled.
Send application materials to: Mr. Terry Hooper, Faculty/Staff Recruitment Office, SUNY College at Brockport, 409 Allen Administration Building, 350 New Campus Drive, Brockport, NY 14420-2929 AA/EOE
Dr. Thomas P. Bonner Department of Biological Sciences SUNY at Brockport Brockport, NY 14420
We have been contacted by Agfa Corporation that they have a small inventory ( approx 30 boxes) of Agfa Scientia 23D56 Electron Microscope film. This film is 3.25" x 4" 100 sheet boxes, with an expiration date of 12/02. Agfa has said they will offer the film at a discount, but prefers to sell it in one bundle. If anyone is interested please contact me directly.
PS- If this is an improper use of this forum, I apologize. It seemed like the quickest method to distribute this information to those who may be interested.
Sincerely,
George
George Laing National Graphic Supply v:(518) 438-8411 X3109 v:(800) 223-7130 x3109 USA f:(800) 832-2205 email: scisales-at-ngscorp.com
We are about to retire a full size refrigerator which had been used to store osmium in both crystalline and liquid form for years. Our Safety people are asking us to clean/decontaminate it before they remove it from the lab. Is there any way to clean this ?
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
If anyone is interested, I have a Narishige Joystick Micromanipulator, model number MN151, for sale. I am asking $1300 or best offer. It has been used very little and it is still in the box.
If you are interested please e-mail me at j-kleier-at-northwestern.edu, off the list serve.
Morning Nami, While the dimensions are somewhat less than optimal, the following might offer two fruitful paths to try, and I emphasize might. I am reminded of an old, tried-and-true, AND direct approach which might help you quickly. Freeze drying of the unfixed organisms. Once the organism is dried, retrieval of the statolith should be facilitated via microdissection as long as it is not fractured by the freezing and drying procedure. While the above will probably enable isolation of the statolith, if I were anxious to study a statolith (with those dimensions) and in ascidian larvae, I would NOT try to isolate it, I would try to expose it in situ. The easiest method by which one might accomplish that is to deep freeze the entire structure, then cleave it with a single-edged razor blade, then look at it directly. If some of my preparations exposed an intact statolith, I would seek to isolate it after I had viewed it with the SEM and EDS/WDS. For TEM, I would process the structure in situ and thin section it as I would a piece of bone. Getting back to freezing and fracturing, with the proper equipment (VPSEM w/cleaving and cryotransfer attachment) you might be rewarded with much valuable data. In the absence of such equipment, I would concentrate first on the statolith which doesn't have to be preserved in the same manner as surrounding tissue. Now, how to accomplish this without expensive equipment. Since best fractures occur at lower temperatures, you might place a piece (brick) of steel (1" thick) in a small SS pan (NOT shallow) which is surrounded by the foam filler one can acquire at Home Depot as an insulator. Pre-cool the steel by pouring LN2 first over, then maintain it around the steel brick. Deep freeze the larva or the statolith, place it on the brick in a drop of an appropriate organic with a low freezing point to bind it to the brick, and cleave the frozen statolith with a pre-cooled single-edge razor blade held in a cooled pair of pliers held by your safely gloved hand. Tap the back of the razor lightly, though simply putting it down might be sufficient, to create a fracture, and hope for the best. NOTE: A member of my undergraduate class suffocated (and died) in an evaporated LN2 atmosphere. Please be careful how you construct an open LN2 system with which you must become somewhat intimate. Also, there are technical concerns beyond safety with which you should be familiar, so I recommend you 'study up' on the subject of freezing biological material for microscopic study before you proceed (if you decide to try this). You have asked for an approach, and I have suggested two. While the confocal might very well provide you with some interesting information about the hemisphere of surrounding tissue on the objective side of the organism, there is far less chance of acquiring any internal information since the statolith is not transparent and will thus be reflective no matter how you embed it. Good luck,
Fred Monson
} From:
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street
West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
To:
} Nami Choe } Ocean Science Centre } Memorial University of Newfoundland } St. John's, NF } A1C 5S7 Canada } phone:(709)737-3247 } fax:(709)737-3220 } } } } }
We sometime document our immunofluorescent images using regular SLR camera and Fujichrome 400 slide film. The problem is that the red fluorescent signal (568nm) did not show up bright on final developed film, However, the green (488) and blue (350nm) ones are fine. We use Olympus BX51 microscopy with a filter set of wide band (U-MWIY2)for red fluorescence.
Since the red signal is perfectly fine through binocular tube, I suspect that either the problem is caused by development (chemical) or film itself.
Any help will be highly appreciated.
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4.214 Dallas, TX 75390-9111
If there is black discolouration inside, it is due to insoluble osmium dioxide. You can possibly remove it by sponging with diluted hydrogen peroxide. This will reoxidise the dioxide to the tetroxide which is more soluble and can be washed off. Of course, in doing this, you will be exposed to osmium tetroxide solution and vapour, so you need protective clothing.
Otherwise you have to scrub off the dioxide with some kind of abrasive cream cleaner (do you have JIF?). The dioxide is inert and relatively harmless.
BTW adding hydrogen peroxide dropwise to osmium fixative which has gone dark will reoxidise it to the tetroxide (and water) and restore it to useful condition. Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
I feel somewhat out of parameters on this list; I have had some thoughts, however, about this topic.
I consider the work of the artist to open our eyes to new realities, to new ways of seeing ourselves, our experiences, the world around us. The artist is concerned to enhance our understanding of the world The physical product of the artist's work are not art, but artifacts. I think this definition of art is problematical, since there obviously is some sense of art work as an aethetic object as well.
Is the work of the scientist not also to shed new light that will enhance our understanding of the natural world?
Both are also concerned with sharing their work. Even the scientist who is cloistered in his corner of the laboratory must eventually be concerned to share new knowledge with others.
I wonder whether graphic data displays (as analytical tools) are objects of art? They can certainly be admirable.
The scientist is concerned with knowledge, with factual truth. This is a difference.
Another thought. I once read, though I don't remember where, a "Balinese saying." I cannot verify its authenticity:
We have no art. We do everything the best we can.
Is scientific writing a form of literary art?
Thank you for allowing me to express these disconnected thoughts.
Alan Davis Marianas High School Saipan, N. Mariana Islands adavis-at-saipan.com
I have just seen the new Nikon Super Coolscan 8000ED demonstrated. It was certainly very impressive. The 4000dpi and the density range of 4.2 make it a great scanner for EM negatives. I'm at present using a LeafScan 45 and this new Nikon was equal to or better than it in all areas and at a fraction of the cost. I have only a couple of reservations about it. The only negative holders that we saw demonstrated would not hold a standard (3 1/4 x 4 in , 8.8 x 10.2cm) TEM negative. We were forced to cut down the negatives. Has anyone been able to construct an adapter to take a full negative? I'm not too worried about the fact that it won't scan the entire area of the negative but don't like the idea of having to cut up negatives. It also seemed that the construction of the negative carriers did not appear to be very solid. I worry that in our multi user centre they may not last very long.
I'd be keen to hear from anyone who has experience with this scanner
Thanks
Rick --
=================================================== Rick Webb Senior Research Officer Department of Microbiology and Parasitology and Centre for Microscopy and Microanalysis University of Queensland 4072 Australia
Mel I purchased some ruthenium tetroxide about a year ago, but the stock went off quite quickly, and the ampoules are now full of a black precipitate. Can I titrate this against H2O2 in the same way ? Chris
} } BTW adding hydrogen peroxide dropwise to osmium fixative which has gone } dark will reoxidise it to the tetroxide (and water) and restore it to } useful condition. } Dr. Mel Dickson, } Deputy Director, The Electron Microscope Unit, } Adjunct Associate Professor, School of Microbiology & Immunology } The University of New South Wales } UNSW SYDNEY 2052 } Australia. } Phone +612 9385 6383 Fax +612 9385 6400 } http://srv.emunit.unsw.edu.au/ }
Most of us work with liquid nitrogen at some stage. Could you tell us more about the circumstances in which someone suffocated. I have up to now assumed small (1L) liquid volumes in a lab were safe. Last year a scientist died in Scotland. As far as I can tell a large volume of liquid was involved.
Dave
On Tue, 23 Oct 2001 12:18:35 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Morning Nami, } While the dimensions are somewhat less than optimal, the following } might offer two fruitful paths to try, and I emphasize might. } I am reminded of an old, tried-and-true, AND direct approach which } might help you quickly. Freeze drying of the unfixed organisms. Once the } organism is dried, retrieval of the statolith should be facilitated via } microdissection as long as it is not fractured by the freezing and drying } procedure. } While the above will probably enable isolation of the statolith, if } I were anxious to study a statolith (with those dimensions) and in ascidian } larvae, I would NOT try to isolate it, I would try to expose it in situ. } The easiest method by which one might accomplish that is to deep freeze the } entire structure, then cleave it with a single-edged razor blade, then look } at it directly. If some of my preparations exposed an intact statolith, I } would seek to isolate it after I had viewed it with the SEM and EDS/WDS. } For TEM, I would process the structure in situ and thin section it as I } would a piece of bone. } Getting back to freezing and fracturing, with the proper } equipment (VPSEM w/cleaving and cryotransfer attachment) you might be } rewarded with much valuable data. In the absence of such equipment, I would } concentrate first on the statolith which doesn't have to be preserved in the } same manner as surrounding tissue. } Now, how to accomplish this without expensive equipment. Since best } fractures occur at lower temperatures, you might place a piece (brick) of } steel (1" thick) in a small SS pan (NOT shallow) which is surrounded by the } foam filler one can acquire at Home Depot as an insulator. Pre-cool the } steel by pouring LN2 first over, then maintain it around the steel brick. } Deep freeze the larva or the statolith, place it on the brick in a drop of } an appropriate organic with a low freezing point to bind it to the brick, } and cleave the frozen statolith with a pre-cooled single-edge razor blade } held in a cooled pair of pliers held by your safely gloved hand. Tap the } back of the razor lightly, though simply putting it down might be } sufficient, to create a fracture, and hope for the best. NOTE: A member } of my undergraduate class suffocated (and died) in an evaporated LN2 } atmosphere. Please be careful how you construct an open LN2 system with } which you must become somewhat intimate. Also, there are technical concerns } beyond safety with which you should be familiar, so I recommend you 'study } up' on the subject of freezing biological material for microscopic study } before you proceed (if you decide to try this). } You have asked for an approach, and I have suggested two. While the } confocal might very well provide you with some interesting information about } the hemisphere of surrounding tissue on the objective side of the organism, } there is far less chance of acquiring any internal information since the } statolith is not transparent and will thus be reflective no matter how you } embed it. } Good luck, } } Fred Monson } } } From: } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } } To: } } } Nami Choe } } Ocean Science Centre } } Memorial University of Newfoundland } } St. John's, NF } } A1C 5S7 Canada } } phone:(709)737-3247 } } fax:(709)737-3220 } } } } } } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
LARGE VOLUME, SMALL SPACE - Truck cab in winter. Truck had a leak. Classmate closed windows of cab and simply went to sleep. Very sad and avoidable. I have found graduate students (still alive) in small closed office labs with open pans of LN2. They were unaware of the potential when air circulation was not assured.
Sorry for the unspecified reference.
Fred Monson
} ---------- } From: Patton, David } Sent: Wednesday, October 24, 2001 4:34 AM } To: Monson, Frederick C. } Cc: 'Microscopy Listserver'; 'Nami Choe' } Subject: Re: RE: microdissection before SEM/TEM } } Most of us work with liquid nitrogen at some stage. Could } you tell us more about the circumstances in which someone } suffocated. I have up to now assumed small (1L) liquid } volumes in a lab were safe. Last year a scientist died in } Scotland. As far as I can tell a large volume of liquid } was involved. } } Dave } } } On Tue, 23 Oct 2001 12:18:35 -0400 "Monson, Frederick C." } {fmonson-at-wcupa.edu} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Morning Nami, } } While the dimensions are somewhat less than optimal, the following } } might offer two fruitful paths to try, and I emphasize might. } } I am reminded of an old, tried-and-true, AND direct approach which } } might help you quickly. Freeze drying of the unfixed organisms. Once } the } } organism is dried, retrieval of the statolith should be facilitated via } } microdissection as long as it is not fractured by the freezing and } drying } } procedure. } } While the above will probably enable isolation of the statolith, if } } I were anxious to study a statolith (with those dimensions) and in } ascidian } } larvae, I would NOT try to isolate it, I would try to expose it in situ. } } The easiest method by which one might accomplish that is to deep freeze } the } } entire structure, then cleave it with a single-edged razor blade, then } look } } at it directly. If some of my preparations exposed an intact statolith, } I } } would seek to isolate it after I had viewed it with the SEM and EDS/WDS. } } For TEM, I would process the structure in situ and thin section it as I } } would a piece of bone. } } Getting back to freezing and fracturing, with the proper } } equipment (VPSEM w/cleaving and cryotransfer attachment) you might be } } rewarded with much valuable data. In the absence of such equipment, I } would } } concentrate first on the statolith which doesn't have to be preserved in } the } } same manner as surrounding tissue. } } Now, how to accomplish this without expensive equipment. Since best } } fractures occur at lower temperatures, you might place a piece (brick) } of } } steel (1" thick) in a small SS pan (NOT shallow) which is surrounded by } the } } foam filler one can acquire at Home Depot as an insulator. Pre-cool the } } steel by pouring LN2 first over, then maintain it around the steel } brick. } } Deep freeze the larva or the statolith, place it on the brick in a drop } of } } an appropriate organic with a low freezing point to bind it to the } brick, } } and cleave the frozen statolith with a pre-cooled single-edge razor } blade } } held in a cooled pair of pliers held by your safely gloved hand. Tap } the } } back of the razor lightly, though simply putting it down might be } } sufficient, to create a fracture, and hope for the best. NOTE: A } member } } of my undergraduate class suffocated (and died) in an evaporated LN2 } } atmosphere. Please be careful how you construct an open LN2 system with } } which you must become somewhat intimate. Also, there are technical } concerns } } beyond safety with which you should be familiar, so I recommend you } 'study } } up' on the subject of freezing biological material for microscopic study } } before you proceed (if you decide to try this). } } You have asked for an approach, and I have suggested two. While the } } confocal might very well provide you with some interesting information } about } } the hemisphere of surrounding tissue on the objective side of the } organism, } } there is far less chance of acquiring any internal information since the } } statolith is not transparent and will thus be reflective no matter how } you } } embed it. } } Good luck, } } } } Fred Monson } } } } } From: } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging(CASI) } } West Chester University of Pennsylvania } } Schmucker Science Center II } } South Church Street } } } } West Chester, PA, 19383 } } eMail: fmonson-at-wcupa.edu } } http://darwin.wcupa.edu/casi/ } } } } To: } } } } } Nami Choe } } } Ocean Science Centre } } } Memorial University of Newfoundland } } } St. John's, NF } } } A1C 5S7 Canada } } } phone:(709)737-3247 } } } fax:(709)737-3220 } } } } } } } } } } } } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
Listers, If anyone has available for sale a manual single or double-tilt specimen holder fitting the Philips CM 10/12 series, please contact me with a price quote and condition of holder. Thanks, Winston ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor 10/24/2001 10:29 AM Cannon Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 (Ship to: 1000 Blythe Blvd ) Fax: 704-355-0589 Charlotte, NC 28232-2861 (Ship to: 28203 ) Winston.Wiggins-at-CarolinasHealthCare.org {mailto:WWiggins-at-Carolinas.org} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ "Vocatus atque non vocatus, Deus aderit." - C.G.Jung
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I'm not sure exactly what info you're looking for, but we have adapters for the Coolpix to mount to just about any microscope. On the Nikon microscopes we have 3 options: a 23 mm phototube, a 30 mm phototube and a 38 mm ISO photoport. Give me a call and we can go over the specifics of your scope.
Tom Tottleben wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } To: Great Minds in the Halls of Science Wisdom! } } I realize this topic has been hammered out in this forum a few times } (for everything except the Nikon Optiphot superwide } trinocular) ......can someone direct me to any information that would be } helpful? ("I can do this .......yeah!!!") Thanks, } } Tom Tottleben } Tottleben Scientific Co } PO Box 24 } 104 Burns Farm-W.Ct. } Edwardsville, IL. 62025 } 618 656 9008 Office } 618 656 9599 Fax } 618 558 9008 Cell } 800 283 9997 Orders } tomtot-at-charter.net } www.tscmicroscopes.com
hello all! for those who work (or starting to work, that`s my case) on Bain tissue research: I`m looking for an rat/mouse antibody anti-NCAM and anti-PSA-NCAM. I recently purchased two antibodies from the Developmental Studies Hybridoma Bank of the U of Iowa, but those abs are anti-embrionic form of rat. I am looking for an `adult`rat/mouse form.
Please, I would appreciate your help
:)
Valeria L.Burgos Instituto de Ciencias Bàsicas y Medicina Experimental Hospital Italiano de Buenos Aires - ARGENTINA Tel: 54-11-9590200 extension 8919
_________________________________________________________ ¿Lo probaste? Correo gratis y para toda la vida en http://correo.yahoo.com.ar
I will appreciate to get any information on alignment of ion sources in Gatan DuoMill
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
I am wondering if there are folks who have used the Panasonic GP-US502 1/2" 3CCD camera and its successor the GP-US522 ?
We are thinking of integrating them on some microscopes.
Have you found GP-US502 sensitive enough to capture fluorescence images ?
Criticisms ? Prices Paid ? Thoughts ?
Thanks,
Sincerely,
Ed Monberg, General Manager LMDC 3101 Whipple Road, Union City, CA 94587 510-429-1060, Fax 429-1065, Cell 510-427-0115 WEB CATALOGUE: http://www.lasermotion.com ---------------------------------------------
To All who are concerned about my earlier LN2 reference.
In an earlier recommendation to the originator of the question about statoliths, I cautioned in my suggestions that a friend had died in an LN2 accident. Apparently there is some concern that I was referring to a death by a small amount of LN2. No one knows what volumes were involved, but the next paragraph should clarify the matter.
I didn't say that my friend died from a liter or, I think, suggest that the young lady could. My fraternity brother did die from asphyxiation from a nitrogen leak while he was trying to fix it in the closed cab of a tanker truck - according to the company AND the coroner at the time. We never could figure out why he was working in the cab or how there could have been a leak to fix in such a location. I even asked if it was the cab or the tank, and it WAS the cab. [A pressure line to the cab to monitor pressure in the tank? In their O2 carriers as well?????] All other info was lawyer'd quiet!
I never had a problem with a liter either, but I have recommended that students not work alone with any amount of LN2 in closed rooms with poor or no ventilation even if I couldn't specify the specific danger. I've never felt even drowsy either, but I haven't dealt with large-volume evolutions.
I did have an assistant once who loaded a 20L dewar in a small unventilated closet in which the 160L dewer was kept. When she came out, she claimed to feel faint. When I checked the dewar, it had lost half its contents in two days, and there appeared to be a leak in the siphon valve. I had it moved to a corner in a larger, well-ventilated room once the valve was replaced. Never had a problem thereafter, even when there were leaks.
In the final analysis, however, when I suggest the use of LN2 to people I don't know who work in environments with which I'm not familiar, I want a little "CYA" on my side.
Sorry I raised a red flag by an obscure reference to death. Perhaps it was ill-advised or at least poorly represented. I will not forget the concerns.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
Dear Listers I wish to sincerely thank everyone who responded to my question.This has opened more doors for me to explore.I am grateful for your generosity, in particular providing me with references and email contacts. Kind regards Judi Bowden
Roy, three easy ways to tell the difference between Si and GaAs:
1) GaAs is not as strong as Si. So GaAs wafers are generally thicker than Si wafers of the same diameter and are easier to break. 2) GaAs is a lot denser than Si. So if you have two wafers the same diameter but one is noticeably heavier (also because of (1)), it's GaAs. 3) GaAs cleaves on (110) and Si cleaves on (111). If your substrate has been cleaved up and has a flat edge perpendicular to the top surface, it's GaAs. If the edge orientation varies and is rarely perpendicular, it's Si.
Hope this helps!
Richard
} From: Beavers, Roy } Sent: Tuesday, October 23, 2001 4:44 PM } To: Microprobe List (E-mail) } Subject: Ni Films } } List, } } You have help me with issues before, which is greatly appreciated, so I } would like to ask help on the latest problem I have been working on. I } have } been looking at some 500 angstrom Ni films on Si substrates trying to } determine if there is any W contamination in the film. } } At first I thought I would use an beam of ~7KeV to keep my analysis within } the film as much as possible but I realized I would probably not see the } Ni } or W X-ray lines with this set up so I decided to go with a beam of 15KeV } which is where I usually run just to see what came up. } } Next I checked a Ni and W standard to see what lines I would get. This } looked o.k. so I proceeded to check a sample of the source material and as } expected I got a match for Ni and only Ni. } } Going to the film samples I expected to see Ni and I did, also I see no W, } but I did expect to see Si because of the higher beam energy but I did } not. } This is my first question why no Si lines? } } What I did see on the film samples that was unexpected was what I believed } to be lines for Ga and As. This may answer my question as to why no Si } signal but I want to check my thoughts on this before I tell my customer } they have deposited films on the wrong substrates. } } Any thoughts again will be greatly appreciated. } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu
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I'm looking for advice or anecdotes about moving heavy SEM equipment. I'm hoping to take possession of an old SEM, whose column base is 33"x33" and may weigh as much as 1,200 pounds. The console is wider and may weigh only 300 pounds.
If I could avoid $1,000 on professional movers and wrestle it with a few strong guys, that would be great, but there are other questions in my mind.
Regardless of who moves it, what considerations must they take regarding the sensitive nature of the equipment? Certainly sensitive bits like filaments should be removed and handled separately, right?
And what about the requirements for trucking? If it's going to spend two or three hours on the interstate, will any sort of truck do the job?
Hi All, To add to Fred's line about potential hazards involved with large volumes of LN2... Last spring, during the final stages of installing a new MRI facility here at the hospital a young man from the company doing the installation was killed by a large volume LN2 leak. I guess the coils of those beasts are cooled and there was a leak in one of the cooling lines and the room was filled with N2 gas...he was asphyxiated immediately. As long as our tanks & valves don't leak, the small volumes we actually deal with on a daily basis are probably not a problem, but I instruct everyone to work in a large room with good ventilation, or to leave the door open. I've had the ventilation in my EM room checked, since that is where my tank is stored.
lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
We are thinking to buy softwares which are good in both high resolution TEM image simulation and dynamical SAD and CBED diffraction simulation.
Can anyone recommend us some good ones?
Best Regards,
Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
I have a transformer for a Bausch and Lomb ring illuminator that I wish to donate to any public institution willing to pay the shipping charge ($ 5-10). I believe this works with the fluorescent ring illuminator for the stereozoom series. The electrical specifications are "PRIMARY: 115 V AC-60 cycles SECONDARY: 750 V 45 milliamps". It appears to be in good working condition (i.e. my voltmeter reads a 750 V output). I can send a digital image on request. Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Rick, Many of our customers are very happy with the 8000ED. I have looked into putting TEM negs into it and it could be done by modifying one of the 120/220 film holders. The holders have a raised lip to keep the film in the channel. I believe these could be removed and the film could then just extend out past the scan opening. I have not been able to try this as demand for the scanner has been very high. Another excellent scanner for TEM negs is the Agfa T2500 Duoscan. While lower in resolution(2500dpi optical) it has a glassless carrier design that will enable scanning of an entire TEM negative. It also will scan reflective originals.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
-----Original Message----- } From: Rick Webb [mailto:r.webb-at-mailbox.uq.edu.au] Sent: Wednesday, October 24, 2001 6:01 PM To: Microscopy Listserver
Hi
I have just seen the new Nikon Super Coolscan 8000ED demonstrated. It was certainly very impressive. The 4000dpi and the density range of 4.2 make it a great scanner for EM negatives. I'm at present using a LeafScan 45 and this new Nikon was equal to or better than it in all areas and at a fraction of the cost. I have only a couple of reservations about it. The only negative holders that we saw demonstrated would not hold a standard (3 1/4 x 4 in , 8.8 x 10.2cm) TEM negative. We were forced to cut down the negatives. Has anyone been able to construct an adapter to take a full negative? I'm not too worried about the fact that it won't scan the entire area of the negative but don't like the idea of having to cut up negatives. It also seemed that the construction of the negative carriers did not appear to be very solid. I worry that in our multi user centre they may not last very long.
I'd be keen to hear from anyone who has experience with this scanner
Thanks
Rick --
=================================================== Rick Webb Senior Research Officer Department of Microbiology and Parasitology and Centre for Microscopy and Microanalysis University of Queensland 4072 Australia
To All: I've been wondering whether to respond or not but here is a situation that happened in Vermont about 10 years ago. An artificial breeder was asphyxiated and died when he fell asleep in the pickup bed of his truck. He had his semen in a 5 - 10 gal LN2 Cryogenic Dewar that was in the bed of his pickup truck. I don't think he had a cab over the bed. The conditions were; warm/hot afternoon in Vermont (~80 degrees) cold for Texas. I believe the Breeder wanted to take a nap in the back truck bed. The Cold Nitrogen filled the truck bed and deplaced the Oxygen. The winds were minimal. He was overcome by a relative small amount of LN2 but there was a time factor. I believe they found him a few hours later. Beer may have also played a role.
Bottom Line: Don't sleep on the floor of the Lab if your using LN2
Dave Campbell IBM Micro-Electronics Essex Junction, Vt.
P.S. I was introduced to the Darwin Awards on this forum, but haven't seen any in a few years. Are there any updates? Please Respond off line.
I would not try to guess on this one. The only real solution is to have an oxygen monitor. They can be a pain because they are now required to be set at our facility to 19.5% for the alarm, so small spills can trip them. They also require weekly maintenance. I'd rather have the small annoyance of false alarms than the one time chance of being dead.
At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould 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 add to Fred's line about potential hazards involved with large } volumes of LN2... } Last spring, during the final stages of installing a new MRI } facility here at the hospital a young man from the company doing the } installation was killed by a large volume LN2 leak. I guess the } coils of those beasts are cooled and there was a leak in one of the } cooling lines and the room was filled with N2 gas...he was } asphyxiated immediately. } As long as our tanks & valves don't leak, the small volumes we } actually deal with on a daily basis are probably not a problem, but } I instruct everyone to work in a large room with good ventilation, } or to leave the door open. I've had the ventilation in my EM room } checked, since that is where my tank is stored. } } lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
Dear John, About two years ago I purchased a used SEM across the country and the movers only charged me about $500 to move it all that way. I chose a firm that delivers new SEMs, so they know what to do. You might ask your nearest SEM service provider for references. The microscope must be disassembled for shipping and the shipping bolts put in to stop things from shaking free. There may be some instructions with the documentation but at the least: 1. Remove the gun, cable and the connection into the high voltage tank and wrap up separately. Remove and box rotary pumps, hoses, power supplies and other connected pieces. 2. Label and disconnect all wires that stretch between the two sections. 3. Bolt down the column section. My moving company did not box the SEM, just put it on two pallets and wrapped it in plastic and moved everything, as much as possible, with a fork lift. Two strong men had no problem maneuvering the column section off the pallet and into the right place. If you think the column section is too heavy, you can probably take the lenses off, as well. This is a professional job to do right and IMHO the risk is worth getting an SEM service professional to pack it up and a scientific instrument moving company to move it is well worth what they charge. Make sure they have an "air-bed" truck for moving. At 07:23 AM 10/25/01 -0500, you wrote: } } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I don't know anything about the stuctures you mention; however if you could work from a slice of tissue (e.g., for Xray), check out these publications:
Howell DN, Miller SE. Identification of viral infection by confocal microscopy. 1999. Methods in Enzymology. Academic Press, 307:573-91.
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.
Briefly, we make "vibratome" slices; stain them with propidium iodide, examine them by confocal microscopy for unusual areas, and then cut out the area of interest. Alternatively, if you have an antibody against the structure of interest, you can use a fluorochrome to label them. In t his manner, we've been able to locate a single virus-infected cell by confocal and then find that same cell by EM.
Good luck, Sara
On Mon, 22 Oct 2001, Nami Choe wrote:
} Date: Mon, 22 Oct 2001 15:37:43 -0230 (NDT) } From: Nami Choe {c56nc-at-morgan.ucs.mun.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: microdissection before SEM/TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopy members; } } Hello. I am a marine biology graduate student at Memorial University of } Newfoundland, Canada. One of my research topic is to study the } microstructure of the statolith in the pelagic marine tunicate (Larvacea), } Oikopleura vanhoeffeni. The statolith is a small cylindrical calcium } build up (10 micrometer diameter, 20 micrometer height), attached to the } brain of the tunicate. This bony structure is contained in the } membraneous statocyst and probably works as a gravity detector. } I would like to isolate this structure for SEM and TEM analyses. After } this, Xray microcroprobe and other procedures will follow. } } Dissecting the brain is easy but taking out the statolith from the } statocyst is the difficult part. Visualizing the statolith can be done by } calcium specific stain, alizarin red. I have tried to dissect the } statolith out by using fine insect pins ( { 20 micron tip size), but the } pins are too large. Doing this under dissecting scope is impossible since } 4x magnification does not allow me to do such microdissection. } } I tried to dissolve the statocyst with bleach and hydrogen peroxide to } isolate inorganic statolith. However, after this procedure, I can no } longer visualize the statolith since alizarin red stain dissolves } instantaneously in these chemicals. } } Sonication was also done to rupture the statocyst and pop the statolith } out but it did not work. } } Do you have any suggestion on how I can separate this structure? } Sincerely, } Nami Choe } ____________________________________________________________________________ } } Nami Choe } Ocean Science Centre } Memorial University of Newfoundland } St. John's, NF } A1C 5S7 Canada } phone:(709)737-3247 } fax:(709)737-3220 } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Would the users of the SymParter attachment for the LKB knifebreaker care to comment on how useful it is? If you want to do it anonymously, e-mail me directly and I will post the replies without attribution. Thanks, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
on 10/25/01 5:23 AM, John Foust at jfoust-at-threedee.com wrote:
} I'm looking for advice or anecdotes about moving } heavy SEM equipment.
I've moved three SEMs, one of them locally, and two of them across-country towed behind (uncovered, but contents well-tarped) rental trailers . My experience has been that hiring SEM engineers is totally unnecessary.
our group works on Transmission Electron Microscopy of semiconducting materials, especially low-dimensional materials for optoelectronic applications. We work on InGaAs/GaAs single and stacked quantum dots and we recently started a new research activity on GaN on sapphire and/or Alumina substrates.
We have a good experience with TEM sample preparation, but we recently decided to buy the Multiprep System, to exploit new methods for TEM sample preparation and to minimize the ion milling time.
We just assembled and calibrated the machine, and we are now in the progress of preparing the first plan-view and cross-sectional samples.
As our machine is one of the few presently available in Italy, we are in contact with Allied to have suggestions and directions.
I should appreciate it very much if those of you who have operated the machine can give us suggestions, hints and procedures to get good quality samples with this nice machine.
Thanks in advance for your help
Massimo
Dr. Massimo Catalano IME-CNR Via Arnesano 73100 Lecce - ITALY phone: +39 0832 391199 fax: +39 0832 325299 email: massimo.catalano-at-ime.le.cnr.it
David this is how it should be done, but there is no point in using a monitor if it is disabled. A feature of the 1999 Scottish fatality was that the deceased habitually disabled the oxygen monitor. Having said that, we have operated without an oxygen monitor for years. Now may be a good time to buy one .... The circumstances of the Scottish fatality were somewhat unrepresentative of lab conditions in that having become unconscious, the victim was exposed to an effectively endless stream of nitrogen delivered from a large-volume dewar. My department responded to that incident by making it a rule that the person delivering nitrogen from a storage dewar should always be accompanied by a colleague. That way, there is a chance that someone can raise an alarm if things go pear-shaped.
Chris
} I would not try to guess on this one. The only real solution is to } have an oxygen monitor. They can be a pain because they are now } required to be set at our facility to 19.5% for the alarm, so small } spills can trip them. They also require weekly maintenance. I'd } rather have the small annoyance of false alarms than the one time } chance of being dead. } } } } At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould 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 add to Fred's line about potential hazards involved with large } } volumes of LN2... } } Last spring, during the final stages of installing a new MRI } } facility here at the hospital a young man from the company doing the } } installation was killed by a large volume LN2 leak. I guess the } } coils of those beasts are cooled and there was a leak in one of the } } cooling lines and the room was filled with N2 gas...he was } } asphyxiated immediately. } } As long as our tanks & valves don't leak, the small volumes we } } actually deal with on a daily basis are probably not a problem, but } } I instruct everyone to work in a large room with good ventilation, } } or to leave the door open. I've had the ventilation in my EM room } } checked, since that is where my tank is stored. } } } } lee } } -- } } Leona Cohen-Gould, M.S. } } Sr. Staff Associate } } Director, Electron Microscopy Core Facility } } Manager, Optical Microscopy Core Facility } } Joan & Sanford I. Weill Medical College } } of Cornell University } } voice (212)746-6146 } } fax (212)746-8175 } } } -- } David R. Hull } NASA Glenn Research Center at Lewis Field } Advanced Metallics Branch } Mail Stop 49-1 } 21000 Brookpark Road } Cleveland, OH 44135 } } (216) 433-3281 } fax (216)977- 7132 } david.r.hull-at-grc.nasa.gov } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html }
Even better, forced ventilation so that accumulation is impossible, and an alarm for when the impossible happens.
Ray } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } Organization: Inveresk Cottage } Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } Date: Fri, 26 Oct 2001 08:17:18 +0100 } To: "David R Hull" {David.R.Hull-at-grc.nasa.gov} } Cc: {microscopy-at-sparc5.microscopy.com} } Subject: Re: Potential LN2 Hazard? } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } David } this is how it should be done, but there is no point in using a } monitor if it is disabled. A feature of the 1999 Scottish fatality was } that the deceased habitually disabled the oxygen monitor. Having said } that, we have operated without an oxygen monitor for years. Now may be } a good time to buy one .... } The circumstances of the Scottish fatality were somewhat } unrepresentative of lab conditions in that having become unconscious, } the victim was exposed to an effectively endless stream of nitrogen } delivered from a large-volume dewar. My department responded to that } incident by making it a rule that the person delivering nitrogen from } a storage dewar should always be accompanied by a colleague. That way, } there is a chance that someone can raise an alarm if things go } pear-shaped. } } Chris } } } I would not try to guess on this one. The only real solution is to } } have an oxygen monitor. They can be a pain because they are now } } required to be set at our facility to 19.5% for the alarm, so small } } spills can trip them. They also require weekly maintenance. I'd } } rather have the small annoyance of false alarms than the one time } } chance of being dead. } } } } } } } } At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould 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 add to Fred's line about potential hazards involved with large } } } volumes of LN2... } } } Last spring, during the final stages of installing a new MRI } } } facility here at the hospital a young man from the company doing } the } } } installation was killed by a large volume LN2 leak. I guess the } } } coils of those beasts are cooled and there was a leak in one of the } } } cooling lines and the room was filled with N2 gas...he was } } } asphyxiated immediately. } } } As long as our tanks & valves don't leak, the small volumes we } } } actually deal with on a daily basis are probably not a problem, but } } } I instruct everyone to work in a large room with good ventilation, } } } or to leave the door open. I've had the ventilation in my EM room } } } checked, since that is where my tank is stored. } } } } } } lee } } } -- } } } Leona Cohen-Gould, M.S. } } } Sr. Staff Associate } } } Director, Electron Microscopy Core Facility } } } Manager, Optical Microscopy Core Facility } } } Joan & Sanford I. Weill Medical College } } } of Cornell University } } } voice (212)746-6146 } } } fax (212)746-8175 } } } } } } -- } } David R. Hull } } NASA Glenn Research Center at Lewis Field } } Advanced Metallics Branch } } Mail Stop 49-1 } } 21000 Brookpark Road } } Cleveland, OH 44135 } } } } (216) 433-3281 } } fax (216)977- 7132 } } david.r.hull-at-grc.nasa.gov } } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html } } } } }
I believe that in the case in Scotland the oxygen monitors were either switched off or ignored.
Dave
On Thu, 25 Oct 2001 17:03:58 -0500 David R Hull {David.R.Hull-at-grc.nasa.gov} 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 would not try to guess on this one. The only real solution is to } have an oxygen monitor. They can be a pain because they are now } required to be set at our facility to 19.5% for the alarm, so small } spills can trip them. They also require weekly maintenance. I'd } rather have the small annoyance of false alarms than the one time } chance of being dead. } } } } At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould 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 add to Fred's line about potential hazards involved with large } } volumes of LN2... } } Last spring, during the final stages of installing a new MRI } } facility here at the hospital a young man from the company doing the } } installation was killed by a large volume LN2 leak. I guess the } } coils of those beasts are cooled and there was a leak in one of the } } cooling lines and the room was filled with N2 gas...he was } } asphyxiated immediately. } } As long as our tanks & valves don't leak, the small volumes we } } actually deal with on a daily basis are probably not a problem, but } } I instruct everyone to work in a large room with good ventilation, } } or to leave the door open. I've had the ventilation in my EM room } } checked, since that is where my tank is stored. } } } } lee } } -- } } Leona Cohen-Gould, M.S. } } Sr. Staff Associate } } Director, Electron Microscopy Core Facility } } Manager, Optical Microscopy Core Facility } } Joan & Sanford I. Weill Medical College } } of Cornell University } } voice (212)746-6146 } } fax (212)746-8175 } } } -- } David R. Hull } NASA Glenn Research Center at Lewis Field } Advanced Metallics Branch } Mail Stop 49-1 } 21000 Brookpark Road } Cleveland, OH 44135 } } (216) 433-3281 } fax (216)977- 7132 } david.r.hull-at-grc.nasa.gov } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
John, You didn't state what kind of SEM. I've moved MANY ETECs with a 5x8 covered U-Haul trailer. I built a 10' folding ramp specifically for these low bed trailers and support the center with a couple of scissors jacks. I prefer to transport them under vacuum because the column is small enough to fit fully assembled and doesn't have a whole lot of mass. The optics table needs to be bolted down, as Mary said, and the high voltage supply handled separately, etc. The chief advantage on these is that they're on wheels and one person can do a move.
For systems that aren't on wheels (almost all others) you can rent a Johnson Bar (pry bar with wheels) and a safe jack (pair of 2 wheeled fork lift units that are strapped to the thing you want to move) and do it yourself (with a friend). You might want to consider a truck with a lift-gate and air suspension. Make sure things are well secured before you drive anywhere.
Good luck!
Ken Converse owner Quality Images third party SEM service Delta, PA
John Foust 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 advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John } } } }
My experience is only anecdotal, but highly successful.
About a month ago I moved a JEOL-840 about 80 miles. The scope was donated to the University by a large corporation. It was decommissioned by a JEOL engineer, but it did not appear to have any special preparation other than to have the spring loaded column table bolted down. It was put on a pallet and moved to the shipping department by professional movers. From there on it was strictly an amateur job. I rented a Ryder truck, which I drove myself. Be sure the truck is high enough to do the job. I had to remove an ion pump at the top of the column in order to get the scope through the door of the truck. I unloaded it on my end with the help of one other person and a hydraulic pallet mover. There was a power lift available on both ends to match the height of the truck bed to the height of the loading dock. Two people (middle aged and not particularly hefty) were able to get the scope off the pallet and to its final position, using pry bars and 1/2 in aluminum rods as rollers. I did the installation myself, and it is now working much better than the similar but older scope it replaced.
If you have a few strong, but also smart helpers and some modest equipment, there is no need to fear the job. Work slowly and carefully. Feel free to call me if you want further details.
Dave Wilbur
-- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University voice: 617-627-2163 Fax: 617-627-3443 email: dwilbu01-at-tufts.edu __________________________________
----- Original Message ----- } From: "Paul B. Grover" {pbgrover-at-home.com} To: "John Foust" {jfoust-at-threedee.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 25, 2001 7:30 PM
Rick:
I second the notion that the AGFA T2500 Duoscan is and excellent scanner. My understanding is that the significant internals are made by Microtek which makes the AGFA T2500 nearly identical to the Microtek ArtixScan 2500 (another excellent scanner.) We also have customers quite pleased with these "twin" units. It's also worth noting, from what I've been told, that AGFA is discontinuing its T2500. I don't know if it's out of production yet, but I do know that because of the discontinuation, we've been able to offer good discounting to the customers we've proposed it to. If you choose to pursue the AGFA, (some inventory is still available) your dealer should be able to offer the same.
Good Luck.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045 *Microscopy and Digital Image Analysis for Science and Industry
-----Original Message----- } From: George Laing [mailto:scisales-at-ngscorp.com] Sent: Thursday, October 25, 2001 3:49 PM To: r.webb-at-mailbox.uq.edu.au; Microscopy-at-sparc5.microscopy.com
Rick, Many of our customers are very happy with the 8000ED. I have looked into putting TEM negs into it and it could be done by modifying one of the 120/220 film holders. The holders have a raised lip to keep the film in the channel. I believe these could be removed and the film could then just extend out past the scan opening. I have not been able to try this as demand for the scanner has been very high. Another excellent scanner for TEM negs is the Agfa T2500 Duoscan. While lower in resolution(2500dpi optical) it has a glassless carrier design that will enable scanning of an entire TEM negative. It also will scan reflective originals.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
-----Original Message----- } From: Rick Webb [mailto:r.webb-at-mailbox.uq.edu.au] Sent: Wednesday, October 24, 2001 6:01 PM To: Microscopy Listserver
Hi
I have just seen the new Nikon Super Coolscan 8000ED demonstrated. It was certainly very impressive. The 4000dpi and the density range of 4.2 make it a great scanner for EM negatives. I'm at present using a LeafScan 45 and this new Nikon was equal to or better than it in all areas and at a fraction of the cost. I have only a couple of reservations about it. The only negative holders that we saw demonstrated would not hold a standard (3 1/4 x 4 in , 8.8 x 10.2cm) TEM negative. We were forced to cut down the negatives. Has anyone been able to construct an adapter to take a full negative? I'm not too worried about the fact that it won't scan the entire area of the negative but don't like the idea of having to cut up negatives. It also seemed that the construction of the negative carriers did not appear to be very solid. I worry that in our multi user centre they may not last very long.
I'd be keen to hear from anyone who has experience with this scanner
Thanks
Rick --
=================================================== Rick Webb Senior Research Officer Department of Microbiology and Parasitology and Centre for Microscopy and Microanalysis University of Queensland 4072 Australia
We would appreciate any information/suggestions concerning service providers for a recently acquired Cambridge Stereoscan 240 SEM. To start we would be interested in a routine inspection and maintenance visit. We are located in Southern Connecticut. Thanks very much!
************************************* Christine Caragianis Broadbridge, Ph. D. Associate Professor of Physics Southern Connecticut State University 501 Crescent Street; Jennings 115 New Haven, CT 06515-1355
Hello list, It's been awhile since I needed work done on our Balzers 301 freeze fracture unit, so I'm asking who provides service on these instruments now? Randy Nessler University of Iowa Phone 319-335-8142
Can anyone give me their thoughts on the design of a request form for users of a microscopy core facility. I have done clinical TEM for twenty years but now I've been asked to manage a confocal/TEM core and will be getting a more diverse mix of specimens. What info should I request from the users? The obvious:1)project design- what questions do you wish to answer 2)sample type- cells, tissue etc. 3) fixed or live 4) fixation method 5) probes? 6) embedding method 7) etc. ?????
should there be two separate forms- one for TEM and one for Confocal? or one general form to include both microscopies?
What do you do? Maybe someone can fax me a copy of their labs request form that I can use as a template. Thank you all. Sincerely, Bill Oxberry Health Science Center Brooklyn fax # 718-270-3313
John: It can be done...I managed with the able assistance of a former student to shift a Hitachi 570 LB via a rental truck from the Smithsonian Institute in Washington, D.C. to Binghamton, NY. ` Success is dependent largely on the care taken with dismantling and packing. If possible, consult a service engineer or a firm that services SEMs of that manufacturer. They should be able to give advice on precautions to take prior to dismantling and shipment. Check out the SEM manual. It should give you exact dimensions and weights for each main part. The height can turn out to be all important...but more of that later. Plan on using lots of labeling tape to label connections. Stretch wrap which comes in 5" and 20" width rolls comes in useful. Boxed components should be plastic or stretch wrapped and well padded. There will be water hoses that will have to be disconnected and drained...and perhaps a water chiller to drain. Disconnect the vacuum hose and pack the rotary pump and make sure its kept upright. Disconnect connections between the display and column units. The cable connecting to the gun and high voltage tank should be removed and the ends stretch wrapped. Pack the cables with care to avoid bending too sharply. As with car spark plug leads the insulation could be damaged. Usually the column is suspended on anti-vibration pads. There should be bolts that can be applied to make the column firm for shipment. Ideally the column should shipped under vacuum. If you remove the top gun section, use a blanking plate to seal the rest of the column and rough pump before shipping. For strapping to pallets and to secure to truck interior you will find belt straps and plastic poly strapping come in handy. Strong backs help, but I would advise having the use of a "J" bar and a heavy duty pallet truck. And speaking of pallets... Just as we were on the loading dock and moving the column unit into the truck we discovered to our consternation we were just over one inch too tall to fit through the truck's door. So don't forget to figure in the height of the column PLUS the pallet when selecting a truck with a tall enough loading door. For us, this meant having to break the SEM column vacuum and remove the gun segment which housed the LB6 filament and ion pump, which fortunately was sealed off under it own vacuum. We covered the lower column with a metal plate and had to ship it without being under vacuum. Upon arrival back in Binghamton, NY the SEM had to be temporally stored in a hallway without access to a 208 V,30 AMP power supply. I had to rig up a system to keep the lower column under rough vacuum until the SEM could be properly installed in its permanent position. I had hole drilled into a half-inch thick aluminum plate cover and threaded to fit a pipe with a gas valve shut-off and a barbed end to fit a vacuum hose. This allowed me to rough-pump the column from the top. It was an interesting experience not to mention the coming and going to the Smithsonian Institute during a rather lively time when streets were blocked off during the World Trade Organization protests...but that's another story. Good luck with your move, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
John Foust wrote: } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John
-- ************************************************ Philosophy is the microscope of thought.
Victor Hugo, Book 3 Chapter 3 Les Miserables ************************************************ Laura Rhoads, Assistant Professor Biology Department SUNY Potsdam 44 Pierrepont Avenue Potsdam, NY 13676 315-267-2260 315-267-3170 fax
A postdoctoral position for electron holography studies of GaN structures is available for 14 months from 1 December 2001.
The project makes use of a Hitachi HF2000 FEGTEM equipped with an electron biprism and Gatan Imaging Filter and aims to develop methods of profiling electric fields which exist at interfaces and around defects. A good background in TEM is essential. Experience in electron holography is not essential.
For further details, contact Prof D. Cherns by e-mail, fax or phone as below
David Cherns H.H. Wills Physics Laboratory University of Bristol Tyndall Avenue Bristol BS8 1TL
Tel +44 (0)117 9288702 Fax +44 (0)117 9255624 E-mail D.Cherns-at-bris.ac.uk
If you want better semiconductor and GaN on sapphire samples than you can ever get by polishing and ion milling, may I suggest that you look into using the MicroCleave technique (also known as the small angle cleavage technique). I would recommend that you visit the South Bay Technology web site and check out the PDF files that they have on the kit. You will see an example of GaN on sapphire in one of their PDF files. There is no ion milling damage present at all. That was one of about five samples that was prepared in about two hours with the kit. Their web site is www.southbaytech.com.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it] Sent: Friday, October 26, 2001 2:53 AM To: Microscopy-at-sparc5.microscopy.com
Dear colleagues,
our group works on Transmission Electron Microscopy of semiconducting materials, especially low-dimensional materials for optoelectronic applications. We work on InGaAs/GaAs single and stacked quantum dots and we recently started a new research activity on GaN on sapphire and/or Alumina substrates.
We have a good experience with TEM sample preparation, but we recently decided to buy the Multiprep System, to exploit new methods for TEM sample preparation and to minimize the ion milling time.
We just assembled and calibrated the machine, and we are now in the progress of preparing the first plan-view and cross-sectional samples.
As our machine is one of the few presently available in Italy, we are in contact with Allied to have suggestions and directions.
I should appreciate it very much if those of you who have operated the machine can give us suggestions, hints and procedures to get good quality samples with this nice machine.
Thanks in advance for your help
Massimo
Dr. Massimo Catalano IME-CNR Via Arnesano 73100 Lecce - ITALY phone: +39 0832 391199 fax: +39 0832 325299 email: massimo.catalano-at-ime.le.cnr.it
} The Surface Science Laboratory at AECL Chalk River Labs has an opening for } a professional surface scientist. Anyone interested in applying is } encouraged to contact either Bill Hocking or Ian Muir. This position will } soon appear on the AECL website (www.aecl.ca) under Careers. Please use } this opportunity to apply on-line. } } Ian Bill } ---------------------------------------------- } ----------------------------------------------- } Ian J. Muir William H. Hocking, Ph.D. } Corrosion and Surface Science Branch Section Head, Surface Science } AECL, Chalk River Laboratories Corrosion and Surface Science Branch } Chalk River, Ontario AECL, Chalk River Laboratories } Canada K0J 1J0 Chalk River, Ontario, Canada } ph: (613) 584-8811 ext. 6960 K0J 1J0 } fax: (613) 584-3250 ph: (613) 584-8811 ext. 4651 } email: muiri-at-aecl.ca {mailto:muiri-at-aecl.ca} fax: (613) 584-3250 } email: hockingw-at-aecl.ca {mailto:hockingw-at-aecl.ca} } } } } Surface Science Laboratory } } The Surface Science Laboratory is part of the Corrosion and Surface } Science Branch within the Fuel Channels Division at AECL. The Laboratory } is equipped with a VG Scientific ESCALAB 220i-XL Imaging X-ray } photoelectron Spectrometer (XPS), a CAMECA ims 6F secondary ion } microanalyzer (SIMS), a Physical Electronics SAM 670 scanning Auger } microprobe and two JEOL scanning electron microscopes (SEM). An SEM/EDX } system is available for analysis of low activity samples, whereas highly } radioactive materials are accommodated in a shielded SEM/WDX facility. } The XPS and SIMS instruments are located in modern laboratories designed } for studies of highly radioactive samples and both systems have been } configured for safe handling of such samples. Activities in the Surface } Science Laboratory range from fundamental studies on nuclear materials to } commercial projects and failure analyses with a focus on characterization } of radioactive materials. } } Surface Science Position } } A research scientist is required to conduct studies of diverse materials } degradation and aging problems in the nuclear industry using secondary ion } mass spectrometry (SIMS) and other surface science methods. Applications } will be focused primarily on radioactive samples taken from reactor } components. } } Duties: } * Stress-corrosion cracking (SCC) of feeder pipes and steam-generator } tubes will be the principal area of initial focus, although support for } on-going studies of corrosion and deuterium ingress in pressure tubes will } be important as well. } * Development of expertise in other areas of applied surface science, } through participation in projects that require a multi-technique approach, } will also be expected. This will involve using scanning Auger microscopy } (SAM), imaging X-ray photoelectron spectroscopy (XPS) and scanning } electron microscopy (SEM) with energy dispersive X-ray (EDX) } microanalysis. } * Preparation of proposals for commercial and internal work, and } reporting to customers on such projects, will be key professional } activities. } * } * Qualifications: } * A graduate degree in surface science and a working knowledge of the } practical application of modern surface analytical methods, especially } SIMS, to industrial problems are required. } * Any additional training in, or experience with, electronics, } computers, spectroscopy, metallography, corrosion processes, radioactive } materials, or ultra-high vacuum technology would be an asset. } } The candidate must be a highly motivated individual with good written and } oral communications skills, who will be able to exploit the commercial } potential of the active surface science capabilities at CRL. }
Hi All, I would appreciate some practical help in choosing between the Leica (Reichert) Ultracut R and the Ultracut S on merit alone, notwithstanding considerations of price.
Thanks,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
The National Institute of Standards & Technology (NIST) has many Post Doctoral Positions open. These are offered competitively through the National Research Council (NRC). Within microscopy & microanalysis research areas we have many possible openings described at the web site listed below. The Microanalysis Research Groups at NIST have about twenty full time scientists specializing in the measurement methods in our extensive facilities including SEMs (FEG, EPMA, Environmental,…), AEMs (FEG/LaB6 TEM/STEM/EDS/EELS), Auger probes, SIMS/TOF-SIMS, MicroXRD/XRF, MicroRaman/IR, Synchrotron beam-line with grazing incidence XPS, etc.
We research new and improved microscopy and microanalysis measurement methods and we apply these methods to challenging analytical problems in semiconductor and optoelectronic technology, materials science, environmental and earth science, etc.
The NIST/NRC Post Doc program offers a two-year position at an annual salary of approximately $53,200 with an additional $5,500 for research expenses. The applications are due to the NRC by Jan 15, 2002. This must include a brief proposal and several recommendations.
A candidate must be a U.S. citizen and start work (with their PhD in hand) at NIST between July 2002 and January 2003. So, this is the perfect opportunity for those that are graduating this spring through next fall and others that have received their degree within the last five years. (Please note that NIST/NRC only competes Post Doc positions once a year, unlike some other institutions. )
Please contact me or go to the following site for further info:
We are in the process of ordering 30 student compound light microscopes for an upper level lab course. Our lowest bid ($400.00 below the next lowest) was for Wesco (or Para/Wesco) microscopes. Has anybody had any expierience with Wesco student scopes and can you make any recommendations. Thank you.
Mark J. Grimson
IMPORTANT NOTE: As required by a state of Texas mandate concerning formatting of state employee e-mail addresses, my new e-mail address is: mark.grimson-at-ttu.edu. The old one should continue to work for quite awhile, but you may want to go ahead and make this change in your address book. Thank you.
Mark J. Grimson, Ph.D Electron Microscopy Facility Dept. of Biology, MS 3131 Texas Tech University Lubbock, Texas 79409
One of the best student microscopes are for Meopta ( or Lambda) Czech rep. address is available on www.coleoptera.org in section scientific suppliers.
They have very good optical resolution, and very good design
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
----- Original Message ----- } From: "The Working Boy" {mark.grimson-at-TTU.EDU} To: "MSA" {microscopy-at-sparc5.microscopy.com} Sent: Saturday, October 27, 2001 7:37 AM
Dear all:
I know that somebody asked a question on how to observe paper structure. This is a little different case. I want to observe the structure of pressure sensitive film, Pressurex Film from Fuji. I am not aware enough of the film structure, but I wonder if I could see the ink capsules which normally breaks and provides red inks on being pressurized. If anybody could explain the film structure and/or the sample prep method, I would appreciate it. I am worried that the organic capsule would break and make mess under the e beam in SEM. If I make the film dry completely, would I see the capsule original spherical or like shrunk baloon?
Dear Jondo Yun, Many years ago I had a customer who routinely checked NCR paper for quality control, checking the size and distribution of the ink balls. I believe he just mounted and coated the samples. There was no problem with bursting spheres, etc.
Ken Converse owner Quality Images third party SEM service Delta, PA
Jondo Yun À±Á¸µµ 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 know that somebody asked a question on how to observe paper structure. This is a little different case. I want to observe the structure of pressure sensitive film, Pressurex Film from Fuji. I am not aware enough of the film structure, but I wonder if I could see the ink capsules which normally breaks and provides red inks on being pressurized. } If anybody could explain the film structure and/or the sample prep method, I would appreciate it. I am worried that the organic capsule would break and make mess under the e beam in SEM. If I make the film dry completely, would I see the capsule original spherical or like shrunk baloon? } } Best regards, } } Jondo Yun } jdyun-at-kyungnam.ac.kr } } }
I never used to have any problem, every so often I just sprayed with a multi-solvent household cleaner and the carbon coating on the inside of the bell-jar wiped off easily.
Now I'm using a different bell-jar, one that had been used for Al and that I had to clean up with acids, and I can't budge the carbon resulting from a few useages. I guess that the acid treatment somehow activated the glass surface so that the carbon really stuck.
Two questions:
How can I clean off this stubborn coating?
How can I prevent it from happening in the future?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Ritchie Sims wrote: ======================================================= I never used to have any problem, every so often I just sprayed with a multi-solvent household cleaner and the carbon coating on the inside of the bell-jar wiped off easily.
Now I'm using a different bell-jar, one that had been used for Al and that I had to clean up with acids, and I can't budge the carbon resulting from a few useages. I guess that the acid treatment somehow activated the glass surface so that the carbon really stuck.
Two questions:
How can I clean off this stubborn coating?
How can I prevent it from happening in the future? ======================================================== So far as removing what is already there, probably you are going to have to use some amount of good old-fashioned rubbing and scrubbing, using acetone and lint free cotton wipers. You can't do anything about the past but you can do something about the future.
And that is to use Bell Bright™ Bell Jar Spray, as shown on URL http://www.2spi.com/catalog/supp/supp3.html
A very thin layer of a water soluble polymer is left behind on the glass surface so that when it is time for a cleaning, a good lint-free cotton wiper and water alone, will cause the deposit to lift right off. In addition to saving a lot of hard work, you eliminate the breathing of acetone vapors.
Now for three words of further comment:
1] The formulation is not substantially different from a typical cosmetic hair spray, however it does not have the various additives that would otherwise gunk up a vacuum system.
2] It is a "gas under pressure" which means it is "dangerous goods" from an international shipment point of view, so it is expensive to ship by itself. It can be shipped only by air freight.
3] Disclaimer: SPI Supplies has formulated and sold this product since about 1978, so we have a vested interest in seeing more of it being consumed
I found your name on the website as someone who teaches existing server-side Java courses. My company has developed a new, practical server-side Java course, and we would like you to consider offering it to your students in the future.
You can view the syllabus of this new class that we developed at www.basebeans.com . The web site has the complete details of the Train the Trainer workshop. As technology advances from servlets to JSPs, the next step is towards JSP frameworks. As an instructor, I'm sure you are aware of the constant need to stay ahead of the curve.
If you would contact me at (415) 781-1463, I can tell you more. Thanks, Rene
1 Jiffy or another fine household abrasive. 2 Coat the inside of the belljar with dilute (roughly 1:20) dishwashing detergent. Dry before use. Less carbon will bounce off the coated belljar. So it gets dark faster, but it just about floats off with a bit of hot water. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, October 30, 2001 12:14 AM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi again } } I never used to have any problem, every so often I just sprayed with } a multi-solvent household cleaner and the carbon coating on the } inside of the bell-jar wiped off easily. } } Now I'm using a different bell-jar, one that had been used for Al and } that I had to clean up with acids, and I can't budge the carbon } resulting from a few useages. I guess that the acid treatment somehow } activated the glass surface so that the carbon really stuck. } } Two questions: } } How can I clean off this stubborn coating? } } How can I prevent it from happening in the future? } } } cheers } } rtch } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Version 4.0 of the Image Processing Tool Kit is now available. Free update copies have been sent to all registered purchasers of version 3.0, so if you neglected to send in the original registration card, now would be a good time to do it! New features, etc., are described at http://ReindeerGraphics.com
Am looking for a source to rebuild PGT detector. Crystal and/or the FET needs redone. I know PGT does things different than any of the other guys as far as the detector. Any names would be helpful. Thanks.
Joel McClintock EM Specialist U of Kentucky 859-257-1242
I once cleaned a very stubborn coating on a glass bell jar by using aluminum foil. Just crumple it up and start wiping. It takes time, but it will come off. I recommend that you use a product like bell bright on the glass prior to using the coater. It puts a layer between the glass and the deposit that is easily to take off with a wet cloth.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, October 29, 2001 9:14 AM To: Microscopy-at-sparc5.microscopy.com
Hi again
I never used to have any problem, every so often I just sprayed with a multi-solvent household cleaner and the carbon coating on the inside of the bell-jar wiped off easily.
Now I'm using a different bell-jar, one that had been used for Al and that I had to clean up with acids, and I can't budge the carbon resulting from a few useages. I guess that the acid treatment somehow activated the glass surface so that the carbon really stuck.
Two questions:
How can I clean off this stubborn coating?
How can I prevent it from happening in the future?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} Randy Nessler } University of Iowa } Phone 319-335-8142 } } }
} } Listers, } } Am looking for a source to rebuild PGT detector. Crystal and/or the } FET needs redone. I know PGT does things different than any of the } other guys as far as the detector. Any names would be helpful. } Thanks. }
Hi, Joel
I'm fairly sure that Gresham, of the UK, will rebuild any brand of detector. I think that Jim Nicolino, of X-Ray Optics, Florida, is their US representative.
But why not involve PGT?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I've been asked to look at some polystyrene microspherules in the SEM and would like some advice on how to prepare them. I'm not a biomedical person, so need some help!
The spherules are 5.6 um size with a Lumidin (protein) bonding surface and receptor molecules attached. They're in DI (or a DI solution?), which is apparently the delivery system for the marker molecules. Would they need CPD? If so, I've never CPD'd particles in solution before.... The SEM has hivac and VP modes, plus there's a cool stage.
Any ideas would be highly appreciated! Many thanks, Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
(Dr. Ritchie Sims on "Coating the Bell Jar" asked how to remove stubborn carbon deposits in a bell jar used to coat aluminum, then acid-etched, then reused with carbon.)
In making carbon fibers for composite materials, it is fairly common for manufacturers to activate the carbon surface (so that it will bond to matrix polymer or to a coupling agent) with an oxidizing-acid etch (often sulfuric or nitric). Dr. Sims, what acid(s) did you use, and at what concentration? Was the acid treatment applied again after carbon deposition, or only before? -------Roy
==================================== Roy Arrowood, Associate Professor Metallurgical and Materials Engineering UTEP, El Paso, TX 79968-0520 (915)747-6934 NEW E-MAIL ADDRESS: arrowood-at-utep.edu
I have been asked to demonstrate, by immunohistochemistry, the presence of sialodacryoadenitis virus, a rat coronavirus, in formalin fixed paraffin embedded tissue. Does anyone know of a commercially available antibody against SDAV that works in paraffin?
Unless they are microballoons, I wouldn't think you would need to do much special. I would probably start quick and dirty and get more complicated if necessary. I would try laying some down on a substrate and try crushing a few between glass slides to get a look at the interior to make sure the spheres are solid.
I would gold coat and use high vacuum mode in order to see much detail. I don't know what your clients want to know. We have some here trying to make small spheres and they are usually somewhat interested in the surface so we have to push for all the resolution we can get.
Warren
At 06:07 PM 10/29/01 -0500, you wrote:
} Hi listers, } } I've been asked to look at some polystyrene microspherules in the SEM and } would like some advice on how to prepare them. I'm not a biomedical } person, so need some help! } } The spherules are 5.6 um size with a Lumidin (protein) bonding surface and } receptor molecules attached. They're in DI (or a DI solution?), which is } apparently the delivery system for the marker molecules. Would they need } CPD? If so, I've never CPD'd particles in solution before.... The SEM has } hivac and VP modes, plus there's a cool stage. } } Any ideas would be highly appreciated! } Many thanks, } Dee } } *************************************************************** } Please do not publicly post any of my correspondence without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Peter: I've had to replace a number of parts recently for our Hitachi S 570 LB. All of the items were shipped promptly by Hitachi Instruments Inc. (800) 253-3053. In the future when parts may no longer be available from the manufacturer, I would try asking for help right here on the MSA list serve. Hopefully, someone will be able to come to the rescue. Henry
-----Original Message----- } From: Peter Tomic [mailto:PTomic-at-Anadigics.com] Sent: Monday, October 29, 2001 8:50 AM To: 'Henry Eichelberger'
John: It can be done...I managed with the able assistance of a former student to shift a Hitachi 570 LB via a rental truck from the Smithsonian Institute in Washington, D.C. to Binghamton, NY. ` Success is dependent largely on the care taken with dismantling and packing. If possible, consult a service engineer or a firm that services SEMs of that manufacturer. They should be able to give advice on precautions to take prior to dismantling and shipment. Check out the SEM manual. It should give you exact dimensions and weights for each main part. The height can turn out to be all important...but more of that later. Plan on using lots of labeling tape to label connections. Stretch wrap which comes in 5" and 20" width rolls comes in useful. Boxed components should be plastic or stretch wrapped and well padded. There will be water hoses that will have to be disconnected and drained...and perhaps a water chiller to drain. Disconnect the vacuum hose and pack the rotary pump and make sure its kept upright. Disconnect connections between the display and column units. The cable connecting to the gun and high voltage tank should be removed and the ends stretch wrapped. Pack the cables with care to avoid bending too sharply. As with car spark plug leads the insulation could be damaged. Usually the column is suspended on anti-vibration pads. There should be bolts that can be applied to make the column firm for shipment. Ideally the column should shipped under vacuum. If you remove the top gun section, use a blanking plate to seal the rest of the column and rough pump before shipping. For strapping to pallets and to secure to truck interior you will find belt straps and plastic poly strapping come in handy. Strong backs help, but I would advise having the use of a "J" bar and a heavy duty pallet truck. And speaking of pallets... Just as we were on the loading dock and moving the column unit into the truck we discovered to our consternation we were just over one inch too tall to fit through the truck's door. So don't forget to figure in the height of the column PLUS the pallet when selecting a truck with a tall enough loading door. For us, this meant having to break the SEM column vacuum and remove the gun segment which housed the LB6 filament and ion pump, which fortunately was sealed off under it own vacuum. We covered the lower column with a metal plate and had to ship it without being under vacuum. Upon arrival back in Binghamton, NY the SEM had to be temporally stored in a hallway without access to a 208 V,30 AMP power supply. I had to rig up a system to keep the lower column under rough vacuum until the SEM could be properly installed in its permanent position. I had hole drilled into a half-inch thick aluminum plate cover and threaded to fit a pipe with a gas valve shut-off and a barbed end to fit a vacuum hose. This allowed me to rough-pump the column from the top. It was an interesting experience not to mention the coming and going to the Smithsonian Institute during a rather lively time when streets were blocked off during the World Trade Organization protests...but that's another story. Good luck with your move, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
John Foust wrote: } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John
-- ************************************************ Philosophy is the microscope of thought.
Victor Hugo, Book 3 Chapter 3 Les Miserables ************************************************ Laura Rhoads, Assistant Professor Biology Department SUNY Potsdam 44 Pierrepont Avenue Potsdam, NY 13676 315-267-2260 315-267-3170 fax
I have an InP p-n junction device with Zn-diffused p-region. I would like to stain-etch the cross-section to observe the junction with SEM. Does anyone have a recipe for this stain-etch? Thanks.
Regards,
Judy Zhu, Ph.D. Member of Technical Staff Multiplex, Inc. 115 Corporate Blvd. South Plainfield, NJ 07080 phone: 908-757-8817 x 1167 fax: 908-757-8910 email: jzhu-at-multiplexinc.com
I've been attempting to do immunogold labelling of 4% paraformaldehyde fixed human natural killer cells (pre- and post- embedding). We're trying to label against perforin, but haven't had success. A monoclonal antibody we have labels the protein in cells fixed with methanol, but we need to preserve the ultrastructure of the cells. Is anyone using an antibody against the protein that works well in paraformaldehyde fixed cells? If so, would you please tell me what vendor you purchased it from, and perhaps share your protocol? Thank you.
Edward Haller University of South Florida Pathology Department Tampa, Florida(813)974-9584
I have become aware of a Community College student, Wil Kunkel, at Scottsdale Community College who will be joining this list in the near future. Wil takes some courses at AZ State U as well, and ended up on my door step with some questions re: SEMs.
It seems that SCC was the recipient of an ISI SX40 scanner from a local business which retired the instrument when its principal operator retired. Neither Wil nor the professor who was the receiver of the gift have any background in microscopy, but Dr. Sickafouse turned Wil loose on the scope after it was dumped in the room and somehow, Wil put it together and got it pumping. He next got a beam (with, as far as I can tell, no help) and started to make micrographs! Pretty good for someone with no EM experience, no vacuum system experience, and no help.
When we met, Wil was looking for information as to how he could get better micrographs. I was happy to spend a Saturday afternoon with Wil and Dr. Sickafouse and their merry band of budding microscopists, teaching some basic principles like gun alignment, signal maximization, vacuum cleanliness and protocols, etc. We were able to ascertain that the SEM was quite usable but will require a column liner dusting and cleaning so it can be stigmated and it will require another diffusion pump cleaning and replacement of the silicone based oil they used to begin with. (I hope they don't have too much trouble with contamination from that - it was a reasonable mistake for people with no Ebeam experience to make.)
My purpose here is to introduce Wil as a new microscopist who has no formal background (so his questions won't always be worded the way those of us who have some years in the business would do) but who has a remarkable interest, a willing mind, and a very high level of ability to reason and adapt - how many of us would - successfully- tackle the cleaning and replacement of an oil diffusion pump with no more help that a typical factory user's manual offers? How many of us could have buttoned up an old scope after a trip across town courtesy of amateur movers and got it to pump with no help?
To get to the purpose of this post - I can and have helped with generic questions and all-purpose adjustments, principles and practices, suggestions on writing a protocol so that new users can be trained and use the instrument for various research projects. What I can't do, and what I hope Wil and Dr. Sickafouse can find on the list, is help with their ISI SX40 specific questions and problems. Obviously, they don't have barrels of money and they have done very well already themselves with the nuts and bolts side of things, but there are sure to be questions that other ISI users and/or factory service people can help with much more expediently than can I.
Thanks in advance to all the generous listers who will, as always, I'm sure, help new people make it work.
Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Here at the Central Michigan Biology Department Microscopy Facility we have found our lab not longer requiring the services of an old SEM that takes incredible images.. . . Lots of spare parts - contact me directly: willi1gl-at-cmich.edu
I am forwarding (with permission) a different take on electron microscopy service. This might be an excellent approach for private industry.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Excalibur Mineral Co [mailto:tony-at-excaliburmineral.com] Sent: Tuesday, October 30, 2001 8:27 AM To: tindallr-at-missouri.edu
We are still accepting applications for the position listed below
US-IL-Chicago Northwest-Electron Microscopist -- SEM
UOP LLC, located in northwest suburban Chicago, is an international leader in technology, products and services for the petroleum processing and separation industries. Our R & D Deptartment, which supports fundamental and market-focused R & D, has an immediate need for an Electron Microscopist to work with chemists and engineers throughout the company on structural and analytical characterization issues relating to catalytic, adsorbent, ion exchanger and metallurgy materials.
Technical assignments will include:
- Interaction and collaboration with solid state and inorganic chemists, applications specialists, and technical service personnel in applying appropriate microscopic and data analysis techniques to problem solving in catalytic and adsorbent materials fields. - Interaction and collaboration with existing microscopy group to effectively utilize SEM and EDXS facilities, including training of junior technical staff. - Development and continuation of in-house expertise in current and emerging scanning electron microscopy techniques and applications.
Minimum qualifications include:
- BS/MS with multi-year relevant work experience, or recent PhD in Geology, Chemistry, Materials Science, or Physics (preferred coursework in electron microscopy, solid state chemistry and physics). - Expertise in SEM and electron probe X-ray microanalysis techniques. - Knowledge of sample preparation techniques, including ultramicrotomy. - Excellent team and communication skills.
Knowledge of other microscopic characterization techniques, for example TEM and STEM, and industrial R&D experience desirable but not essential. Above all, we are seeking a motivated individual, who will demonstrate a willingness to learn and openness to acquiring new areas of expertise in characterization, chemistry and materials properties.
We offer a state-of-the-art scientific research environment coupled with exciting career opportunities coupled with a highly competitive compensation and benefits package including medical and dental coverage, 401(k) Plan, Pension Plan and tuition reimbursement.
US employment authorization required.
UOP is an Equal Opportunity Employer
Visit us at www.uop.com
---------------------------------------------------------------------------- ---- Additional Information Position Type: Full Time, Employee Ref Code: MB-01-11
---------------------------------------------------------------------------- ---- Contact Information UOP Internet Resume Manager cro-at-uop.com UOP LLC 25 E. Algonquin Road, P.O. Box 5017 Des Plaines IL 60017-5017 Fax: 847.391-2921
I have examined polystyrene beads under the SEM. It is not difficult to do. I applied a suspension of beads to a solid support with a pipetter, attached the support to an SEM stub, then sputter-coated the beads with gold. Once the sputter-coater pulls a vacuum, your solution will evaporate. I then viewed the beads under the SEM as usual, without any special attachments.
One thing you need to be careful of is losing the beads. Unless they are physically attached to your surface, they will not adhere to it. (Since they are spheres, they roll off, I guess.) If you have a steady hand this will not be a problem. The biggest challenge for me was smoothly inserting and removing the stub with the beaded support from our sputter coater. Several times I took the beads/stub over to another building to use the SEM. This involved walking down several flights of stairs, and I did not lose many beads (i.e., I saw numerous beads under the SEM.)
Hope this helps,
KD
-----Original Message----- } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] Sent: Monday, October 29, 2001 6:07 PM To: microscopy-at-sparc5.microscopy.com
A few colleagues and I have an interest in viewing SEM images of anthrax spores/virus. If someone can point me to a website that may have them in a library accessible to me, or if someone would forward some to me, we would greatly appreciate it.
Just so everyone is aware, this laboratory's principal function is the study of microelectronic failure mechanisms and not in any way related to biological technologies or biological warfare. We're simply curios.
Regards, Peter Tomic Anadigics, Inc. Warren, New Jersey
I am the lab coordinator here at ASUB. We have light microscopes with 100X oil immersion lenses. My supervisor wants me to attend a school of microscopy to enable me to better maintain and repair our microscopes. Do you know of available schools of microscopy that will meet my needs? Thanks, Linda
Rice University invites applications for a Staff Scientist position in the newly established Center for Biological and Environmental Nanotechnology (CBEN). Responsibilities include management of a major electron and probe microscopy facility, training of students in the use of the equipment, day-to-day maintenance of equipment, and participation in the research projects at Rice. The electron microscopy facility includes a JEOL 2010 TEM, JEOL 6320F field emission SEM, Phillips XL30 environmental SEM, Nanoscope AFMs, a Kamica Electron Microprobe and high-resolution XRD. This position requires a Ph.D. in Materials Science, Physics or a related field with specialization on electron microscopy. Preference will be given to the applicant with (1) experience in hands-on operation and maintenance of a range of electron and probe microscopes, (2) prior management experience, and (3) demonstrated research experience involving transmission electron microscopy. Interested persons should apply (include resume, statement of interest, list of publications, names and addresses of at least three references) by November 15, 2001 to:
Dr. Kevin Ausman Center for Biological and Environmental Nanotechnology MS-63 Rice University P.O. Box 1892 Houston, TX 77005-1892
*Rice University is an Affirmative Action/Equal Opportunity Employer*
I know this question was discussed in the past, but anyway. I would need a pretty comprehensive textbook(s)/ reference book(s) covering EM use in biology - possibly with stress on TEM, including sample preparation techniques. Any bright ideas?
Thanks,
Michael Jarnik
-- Michael Jarnik, Ph.D. Fox Chase Cancer Center Electron Microscope Facility Philadelphia
Semantics because this mistake has been making me crazy - ANTHRAX IS A BACTERIUM! This is a particularly important point because the word "virus" scares people silly. When I think of virus, I think of AIDS and Ebolla (sorry it that's misspelled). When I think of bacterium, I think antibiotic. No offense intended, Kristen
At 09:36 AM 31-10-2001 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011
Thanks to everyone for their helpful suggestions. Given below is the summary of all the replies I received regarding disposal of osmium contaminated refrigerator.
****************************
You can make them happy by moving the refer next to a hood and cleaning it with household bleach, the stronger the better. What this does is takes the relatively safe but dark osmium dioxide residue and converts it to the colorless but dangerous osmium tetroxide residue. It is generally impossible to decontaminate the surfaces, just cycle between toxic and nontoxic. The only other way is to rip out the contaminated areas of the interior of the refer and dispose of it in a toxic landfill. I'm sure they will appreciate this for milligram quantities of osmium. *****************************
If there is black discolouration inside, it is due to insoluble osmium dioxide. You can possibly remove it by sponging with diluted hydrogen peroxide. This will reoxidise the dioxide to the tetroxide which is more soluble and can be washed off. Of course, in doing this, you will be exposed to osmium tetroxide solution and vapour, so you need protective clothing. Otherwise you have to scrub off the dioxide with some kind of abrasive cream cleaner (do you have JIF?). The dioxide is inert and relatively harmless. BTW adding hydrogen peroxide dropwise to osmium fixative which has gone dark will reoxidise it to the tetroxide (and water) and restore it to useful condition. ***************************
There is often times a wide dichotomy between what safety people demand and what common logic and good sense suggest. If it is there at all, it would be black, right? Remember that if it was actually in the tetroxide form, it would be colorless and you would not see it, you would smell it, but actually it would disappear by itself because of its high vapor pressure. So if you have areas that are black, it would be the dioxide which is harmless. Just don't put in contact with any oxidizing agents which would reverse the reaction back to the tetroxide. You can clean it off however you think is the most convenient, perhaps with some laboratory Alconox, and then you could collect the reside removed and put it aside for recycling with your other precious metals slated for recyling and recovery. Unfortunately, too many "safety" people are starting to actual believe what they tell people like yourself. A few weeks ago, we had someone like yourself being told the refrigerator had to be sent to land fill and buried in its entirety! *********************************
100% ethanol will reduce the OsO4 to OsO2, and the latter is a harmless oxide.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
One of the particularly good ones is: Bozzola JJ, Russell LD, 1991. Electron Microscopy: Principles and Techniques for Biologist. Jones and Bartlett Publisher (Boston), 542 pp, ISBN 0-86720-126-6.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Wednesday, October 31, 2001 11:15 AM -0500 Michael Jarnik {M_Jarnik-at-fccc.edu} wrote:
} } Listers, } } I know this question was discussed in the past, but anyway. I would need } a pretty comprehensive textbook(s)/ reference book(s) covering EM use in } biology - possibly with stress on TEM, including sample preparation } techniques. Any bright ideas? } } Thanks, } } Michael Jarnik } } } -- } Michael Jarnik, Ph.D. } Fox Chase Cancer Center } Electron Microscope Facility } Philadelphia }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmichael-at-usc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, October 31, 2001 at 12:13:55 ---------------------------------------------------------------------------
Email: dmichael-at-usc.edu Name: Darren Michael
Organization: University of Southern California
Education: Graduate College
Location: Los Angeles, CA
Question: Hi,
I'm trying to find high index of refraction plastic coverslips. I've read about eyeglasses with high index of refraction (1.66) plastic lenses and have been unable to locate a manufacturer of coverslips using the same types of materials. Have you come across them? Do you have any suggestions as to whom to ask?
My department here is looking to acquire interactive scheduling software, preferably for the internet, that will cover all of our microscopes. Users should be able to create an account, manage it (add appointments, edit them, delete them...), and be able to see what time slots are available for the scope they wish to use. Please feel free to give me all suggestions that come to mind as to what software is available that will accomplish this. Thank you all very much.
Chris
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
Just wondering if anyone can give me some idea of the correct procedure for closing and opening both the column and gun valves when venting a Leica 440 SEM. Our manual only talks about closing the gun valve when using a Lab6 filament. The manual also talks about the gun valve 'flashing' when the vacuum is ready, and that at this point, the valve should be re-opened. Both the valves on our machine flash upon closing them and stay flashing until we return the leavers to the open position We are using a Tungsten Filment not a Lab6 and we do not have an Ion Pump on the column. We have the standard Rotary and Turbo pumps.
The manual does not explain when to close or open these valves when using a Tungsten Filament. My question is:- what is the correct procedure for closing the valves and re-opening them if we are using a Tungsten Filament and not a Lab6. At present we are closing the valves upon venting the machine in order to keep some vacuum in the column when the chamber is at air. As soon as we close the valves they flash red. Once we pump the column again, the gun and column valves stay flashing until we return the leavers to the open position.
Maxine Dawes Technical Officer School of Environmental Science and Management Southern Cross University PO Box 157 Lismore, NSW, 2480