Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, January 2, 2005 at 06:56:05 ---------------------------------------------------------------------------
Organization: The Unit for Nanoscopic Characterization, The Hebrew University of Jerusalem
Title-Subject: [Microscopy] [Filtered] MListserver: Need help in imaging of microemulsions
Question: Dear Listers, I need your help in imaging microemulsions of types oil-in-water/water-in-oil with oil drops of 10 to 50 nm size. Do you have any experience in SEM/ESEM/CryoSEM imaging of such objects?
Many thanks to those who replied to my earlier posting regarding CL alignment on a JEOL 840, but please note that it's the CL alignment specifically (the bit you have to do after tearing down the column) that I'm having problems with, not the objective aperture or stigmator adjustment.
I'm still seeking understanding of that part (ie the CL alignment), if anyone has that one sorted.
thanks and all the best for 2005
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 3 08:19:47 2005
If you have access to an SEM with a liquid nitrogen-cooled cryostage and a way of plunge freezing samples and transferring them frozen into the specimen chamber, this would be a good way to do it. I have imaged foamy liquids with great success like this, and the process was very simple.
Briefly, the liquid to be imaged was shaken violently to form the bubbles, then a drop of the foam was put on the specimen holder and plunged into liquid nitrogen. I was using an EMITech cryopreparation unit, so I took the sample out of the LN2 bath under vacuum and transferred it to the fracture and coating stage of the unit. Using a built-in pick, I fractured off a piece of the frozen foam, then coated the sample with gold before transferring it under vacuum to the SEM stage. Micrographs were taken with no problems.
This was actually one of the easiest cryo specimens I ever worked with, and I expect yours would work much the same way. My only concern might be the size of the droplets you want to image, since cryoSEM, in my experience, doesn't have the resolution of "normal" SEM work. If you have any questions, please let me know.
Good luck, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! 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.emc.missouri.edu
-----Original Message----- } From: by way of MicroscopyListserver [mailto:innap-at-savion.cc.huji.ac.il]
Sent: Sunday, January 02, 2005 8:54 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, January 2, 2005 at 06:56:05 ------------------------------------------------------------------------ ---
Organization: The Unit for Nanoscopic Characterization, The Hebrew University of Jerusalem
Title-Subject: [Microscopy] [Filtered] MListserver: Need help in imaging of microemulsions
Question: Dear Listers, I need your help in imaging microemulsions of types oil-in-water/water-in-oil with oil drops of 10 to 50 nm size. Do you have any experience in SEM/ESEM/CryoSEM imaging of such objects?
The new CytoViva system which debuted last month at Cell Bio, would also give you an interesting approach, with much less sample prep.
The system provides both contrast and high resolution. It retrofits easily to existing light microscopes (research stands) and I've seen it work quite well with colloids on the order of 50nm. The neat thing for emulsions is that you can observe them in real time.
If you are interested, take a look at their website : www.CytoViva.com. Although the applications shown there are biological, I've worked with emulsions before and the imaging principles are very similar. I'd also recommend your contacting Dr. Tom Hasling at Aetos (the manufacturer) and ask him to run a sample for you (+1 334-749-0134). If you can't send him something directly, perhaps you can suggest an emulsion system which is similar to yours.
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
Caveat: MME was part of the initial launch team for this product.
At 08:24 AM 1/3/2005, Tindall, Randy D. wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 3 18:42:58 2005
The new CytoViva system which debuted last month at Cell Bio, would also give you an interesting approach, with much less sample prep.
The system provides both contrast and high resolution. It retrofits easily to existing light microscopes (research stands) and I've seen it work quite well with colloids on the order of 50nm. The neat thing for emulsions is that you can observe them in real time.
If you are interested, take a look at their website : www.CytoViva.com. Although the applications shown there are biological, I've worked with emulsions before and the imaging principles are very similar. I'd also recommend your contacting Dr. Tom Hasling at Aetos (the manufacturer) and ask him to run a sample for you (+1 334-749-0134). If you can't send him something directly, perhaps you can suggest an emulsion system which is similar to yours.
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
Caveat: MME was part of the initial launch team for this product.
At 08:24 AM 1/3/2005, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 3 22:38:47 2005
Happy New Year...Hope you are all well and still on holidays instead of being back at work. I wish I was.
Now to my question....This is probably a silly question but its better to ask. Can anyone recommend an etch solution to reveal the grain structure of electro plated Ni. I know of Kalling's No. 2 and Acetic Glyceregia for Ni based alloys. I expect them to work on Ni but just wanted to be sure and find out if anyone can recommend at better solution.
Regards George
George Theodossiou Physicist / Microscopist Microscopy and Microanalysis Laboratory
AMCOR Research and Technology Ph: +61 3 9490 6135 Fax: +61 3 9499 4295 Mobile: 0409 568 840 email: George.Theodossiou-at-amcor.com.au
************************************************************************ CAUTION - This message may contain privileged and confidential information intended only for the use of the addressee named above. If you are not the intended recipient of this message you are hereby notified that any use, dissemination, distribution or reproduction of this message is prohibited. If you have received this message in error please notify AMCOR immediately. Any views expressed in this message are those of the individual sender and may not necessarily reflect the views of AMCOR. ************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 00:47:11 2005
The etchant that I prefer for electoplated nickel is a 1:1 mixture of acetic and nitric acids.
George Theodossiou writes:
} Can anyone recommend an etch solution to reveal the grain structure of } electro plated Ni. I know of Kalling's No. 2 and Acetic Glyceregia for Ni } based alloys. I expect them to work on Ni but just wanted to be sure and } find out if anyone can recommend at better solution. } } Regards } George } } George Theodossiou } Physicist / Microscopist } Microscopy and Microanalysis Laboratory }
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 02:33:17 2005
} From Etchants Database, http://www.kaker.com/Etch/Etch.html :
Material: Nickel and Nickel Alloys Type: Polishing Method: Electropolishing Etchant (Electrolyte): 37 ml H3PO4 (conc.), 56 ml glycerol, 7 ml water. Procedure: 9-10 A/in2, Pt cathode, water cooled. Remarks: Nickel 200. Reference: Metallography, Structures and Phase Diagrams, Metals Handbook, 8th Edition, Vol. 8, ASM (American Society for Metals), Metals Park, Ohio 44079, USA, 1973, p. 137.
Material: Nickel and Nickel Alloys Type: Macro Method: Chemical etching Etchant (Electrolyte): 25 ml nitric acid (conc.), 75 ml hydrofluoric acid (conc.). Procedure: Etching time is 3-10 min. Use fume cupboard. Remarks: Ni-Cr and Ni-Cr-Fe alloys. Reference: H. Modin and S. Modin, Metallurgical Microscopy, Butterworths, London, 1973., p. 390.
Material: Nickel and Nickel Alloys Type: Polishing Method: Electropolishing Etchant (Electrolyte): 37 ml H3PO4 (conc.), 56 ml glycerol, 7 ml water. Procedure: 8-10 A/in2, Pt cathode, water cooled. Remarks: Duranickel 301. Reference: Metallography, Structures and Phase Diagrams, Metals Handbook, 8th Edition, Vol. 8, ASM (American Society for Metals), Metals Park, Ohio 44079, USA, 1973, p. 137.
Material: Nickel Specimens and Nickel Alloys Type: Micro Method: Electro polihsing Etchant (electrolyte): A.) For Ni: 50 % hydrochloric aicd, 25 % water. B.) For Ni alloys: 75 % hydrochloric acid, 25 % water. Procedure: No data. Remarks: Jet electro polishing for discs. Reference: S.Mader, et.al., J.App.Phys., Vol.34, 1963, p.3376.
Henrik Kaker
George Theodossiou wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi All, } } Happy New Year...Hope you are all well and still on holidays instead of } being back at work. I wish I was. } } Now to my question....This is probably a silly question but its better to } ask. Can anyone recommend an etch solution to reveal the grain structure of } electro plated Ni. I know of Kalling's No. 2 and Acetic Glyceregia for Ni } based alloys. I expect them to work on Ni but just wanted to be sure and } find out if anyone can recommend at better solution. } } Regards } George } } George Theodossiou } Physicist / Microscopist } Microscopy and Microanalysis Laboratory } } AMCOR Research and Technology } Ph: +61 3 9490 6135 } Fax: +61 3 9499 4295 } Mobile: 0409 568 840 } email: George.Theodossiou-at-amcor.com.au } } ************************************************************************ } CAUTION - This message may contain privileged and confidential } information intended only for the use of the addressee named above. } If you are not the intended recipient of this message you are hereby } notified that any use, dissemination, distribution or reproduction of } this message is prohibited. If you have received this message in error } please notify AMCOR immediately. } Any views expressed in this message are those of the individual sender } and may not necessarily reflect the views of AMCOR. } ************************************************************************
-- Dr. Henrik Kaker Metal Ravne SEM-EDS Laboratory Koroska cesta 14 SI-2390 Ravne Slovenia Phone: +386 2 870 7076 Fax: +386 2 870 7022 GSM: +386 31 380 875
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 05:20:02 2005
I am wondering whether anybody can help me obtaining microscopical slides of the nervous system of classical experimental animals.
I will be teaching students in a course on histology of the nervous system and am supposed to compare specimens of the human nervous system with the nervous system of classical experimental animals. We have many slides of the human nervous system here at this department, but are lacking slides of other animals. I myself work on insects, so it will be easy for me to prepare slides with ganglia of insects, but for comparison, I shall need slides of other invertebrates (squid, Aplysia, Lymnaea... ?). Does anybody know a source of spare, prepared slides of parts of the nervous system of such invertebrates? Or, does anybody know a source of Aplysia or Lymnaea and have dissection istructions, so that I can attempt to dissect them myself?
thank you
Gerd Leitinger
Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, January 4, 2005 at 05:55:02 ---------------------------------------------------------------------------
Email: neil-at-young8696.freeserve.co.uk Name: Neil
Organization: University of Birmingham
Title-Subject: [Microscopy] [Filtered] TEM School, Antwerp
Question: Hi, just thought I'd see if anyone is attending the winter school at Antwerp this month?
Neil Young Nanophysics Research Laboratory University of Birmingham
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jfrancis-at-powell.k12.ky.us) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, January 4, 2005 at 11:37:24 ---------------------------------------------------------------------------
Email: jfrancis-at-powell.k12.ky.us Name: Jennifer Francis
Organization: Powell County Schools
Education: 6-8th Grade Middle School
Location: Stanton,KY, USA
Question: I am formerly an elementary school teacher, now working with middle school students. I would like to invest in a microscope that we can view on a tv or computer monitor. Can you tell me what equipment I might need to do that? I am not sure what the differences are between a digital microscope, a flex cam, etc. The digital microscopes seem to be less expensive, and I wondering if I connect that to a computer/laptop if I could use a digital projector (already own) to project from there and if that is better than something like the flex cam? Would you have any ideas? Thanks!
============================================== Thrid and Final Notice of Microscopy Listserver DataBase Update: ============================================== Jan 4, 2005
Colleagues, if you have subscribed/resubscribed since Dec. 21st You may ignore the remainder of this message you will already be in the new database. The Listserver will begin using the new database records next week.
======================================= Second Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 28, 2004 ======================================= First Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 21, 2004
Colleagues....
After nearly a dozen years of operation, the master database of the Microscopy Listserver needs a serious reworking and purging. I have done the hard work of restructuring the software model but now the database needs to be "repopulated" with updated/new user information.
The easiest way for me to accomplish this is to simply purge the entire database, and have each of you resubscribe as if you are a new user.
As a result, I'm asking EACH and EVERY ONE of you to visit the following WWW site and fill out the new subscription form. Here is the URL:
You can do this beginning immediately, the changes will be stored and I will implement the new system on or about the second week of Jan. 2005. If all goes well, resubscribing should only take a few minutes of your time, so please do it as soon as possible.
For the balance of 2004 the old database will remain in use, but if you have not resubscribed by ~ the first week of Jan 2005, you will no longer receive Listserver Email as I will cease using the old database and only forward Listserver Email to those individuals who have subscribed and are recorded in the new system.
I'll post this message every Monday through Jan 3, as a reminder and shortly thereafter will transition away from the old database to the new one.
Thanks in advance for your patience and cooperation.
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 16:57:04 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, January 4, 2005 at 08:24:11 ---------------------------------------------------------------------------
Question: My co-worker has a proposal for a massive database of TEM images. Here it is.
I am interested in finding (or creating) an easily accesible transmission electron microscopic image database which will have thousands of images available to preview....without comments or arrows or descriptions on the actual images.... but with the option to retrieve the article or a description of the treatment/species/etc on a separate link. There could be an image preview mode, and a high resolution link and from there a link to the specifics of species and treatment etc and/or an article on PubMed or Medline. The database should be open to the public, and have a mechanism for adding images to the database (in a particular format) through fields just like filling out a form. The sorting of images could be by mesh headings. The databases which I have been able to access do not appear in this format, and there is too much in the way of "other links and descriptors" to allow for a quick visual scan of the images. Does anyone out there have an interest in such a database... and would anyone out there be willing to be co-investigator on a grant to prepare and maintain such a database. Thanks for your responses.
There are alot of possible image databases out there both commerical and public-domain. None that I know of do exactly what you ask, but then that is why you asked for input isn't it?
I'll point you to the public-domain TPM Electronic Notebook at :
http://tpm.amc.anl.gov
As an electronic notebook, it does alot of what you asked for and technically it is simply a database with a WWW based GUI front end. As a Notebook it handles not only text but also images. It is searchable and can be customized with a modicum of effort to incorporate nearly everything you asked about.
If your interested in fiddling, go to the WWW site, then select ENotebook Button, then choose the PUBLIC Notebook and "Enter the ENotebook. Both public and private ENotebooks are available, the difference being simply login requirements.
Each "record" in the Notebook is a seperate page, but the pages can be of unlimited length and data can be added to any individual pages at any time (dated and time stamped of course).
A customizable thumbnail page can be made to show summary images (Table of Contents). Which will give you a preview of multiple pages.
There are some example images in the public notebook. Comment fields, form based input, various file formats supported (JPG/TIF/GIF/BMP/PNG/QT/MOV/MPG/AVI/PDF/DOC/XLS/PPT/ASCII/Binary ) thumbnails and full view images. Links and articles describing the images are appendable to the page, or can be embedded on the page itself.
The ENotebook is (obviously) WWW based and currently runs on a Linux Server. This is part of the TelePresence Microscopy Collaboratory Project at Argonne National Lab. A single server can support multiple private and/or public ENotebooks. In principle the only limitation to the number of images is the size of your Disk Drive.
Rumor has it I know the developer, so get back to me if you have any questions.
Nestor Your Friendly Neighborhood SysOp
} } Email: Stacey.Andringa-at-uc.edu } Name: Stacey Andringa } } Organization: University of Cincinnati } } Title-Subject: [Microscopy] [Filtered] image database } } Question: My co-worker has a proposal for a massive database of } TEM images. Here it is. } } I am interested in finding (or creating) an easily accesible } transmission electron microscopic image database which will } have thousands of images available to preview....without } comments or arrows or descriptions on the actual images.... } but with the option to retrieve the article or a description } of the treatment/species/etc on a separate link. There } could be an image preview mode, and a high resolution link } and from there a link to the specifics of species and } treatment etc and/or an article on PubMed or Medline. The } database should be open to the public, and have a mechanism } for adding images to the database (in a particular format) } through fields just like filling out a form. The sorting of } images could be by mesh headings. The databases which I } have been able to access do not appear in this format, and } there is too much in the way of "other links and } descriptors" to allow for a quick visual scan of the images. } Does anyone out there have an interest in such a database... } and would anyone out there be willing to be co-investigator } on a grant to prepare and maintain such a database. } Thanks for your responses. } } marian.miller-at-uc.edu } } } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 18:17:31 2005
We have an Olympus BX61 that we are using alternating control of from one software package to another. One of the software packages is proprietary and must have the dip switches on the microscope communications hardware set a certain way.
Does anybody know what these switch setting mean so that we can attempt to connect with the other software which may be flexible?
I posted images of the problem at http://www.aecom.yu.edu/aif/temp/sky/index2.htm
Any help would be greatly appreciated.
Thank you.
-Michael ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ **This electronic transmission contains information that is privileged.**
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 18:53:24 2005
On Jan 4, 2005, at 2:09 PM, by way of Ask-A-Microscopist wrote:
} Question: I am formerly an elementary school teacher, now working with } middle school students. I would like to invest in a microscope that } we can view on a tv or computer monitor. Can you tell me what } equipment I might need to do that? I am not sure what the differences } are between a digital microscope, a flex cam, etc. The digital } microscopes seem to be less expensive, and I wondering if I connect } that to a computer/laptop if I could use a digital projector (already } own) to project from there and if that is better than something like } the flex cam? Would you have any ideas? Thanks! } Dear Jennifer, Caroline Schooley is an expert on this; her contact info and the web site for Project MICRO are:
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Project MICRO's purpose is to enhance pre-college education, as stated on the web page. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 4 20:22:03 2005
Hi Stacey We have such a data base at UBC. Dr. Lacey Samuels instigated it as a resource for a Bio200 cell biology course and Joseph Dietz designed it. It just grew from there. Anyone can add images to it, and we encourage users of this facility to "add a useful image." You can access it from my website http://www.emlab.ubc.ca On the left hand side is the link to the BioMedia Database. There is a very easy search engine.
We have the policy that if the images are to be used for educational purposes and non-profit then you can download them for free. If they are to be used for profit, then the copyright stays with the author.
We are at present setting up the link to the protocols pages on the emlab website for the images in the database.
Let me know if you would like to add images to it and need more information. Elaine
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Dr. Elaine Humphrey Director, BioImaging Facility 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-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 07:44:54 2005
Seeing that you are in the Bronx, why don't you just call out to Melville and ask them? They are very freindly and should be able to fax you off the dip switch info.
Corporate Headquarters: Olympus America Inc. 2 Corporate Center Drive P.O. Box 9058 Melville, NY 11747-9058 1-800-645-8160
} } We have an Olympus BX61 that we are using alternating control of from one } software package to another. One of the software packages is proprietary } and must have the dip switches on the microscope communications hardware } set a certain way. } } Does anybody know what these switch setting mean so that we can attempt to } connect with the other software which may be flexible? } } I posted images of the problem at } http://www.aecom.yu.edu/aif/temp/sky/index2.htm } } Any help would be greatly appreciated. } } Thank you. } } -Michael } ____________________________________________________________________________ } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ } **This electronic transmission contains information that is privileged.** } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 11:29:49 2005
You can actually turn nearly any microscope into a digital microscope by attaching a camera to it. Several companies which come to mind which have cameras that can slip right into the place where a normal eyepiece goes are VideoLab and Ken-a-Vision. I'd also suggest seeing if Swift Instruments has anything.
Several years ago, we taught a week-long course for jr. high school and high school teachers (ironically, chemistry and physics) at Miami University (Oxford, OH). Early in the program, we put these small, inexpensive cameras (I think, at that time, that the VideoLab camera only cost about $258 for a direct feed into a video monitor... maybe another $125 for a card to go into a computer, if you wanted to capture images) on the normal, high-school level microscopes and ran experiments on polarized light, diatoms, and contrast techniques (ex: salt crystals under Darkfield and a technique popular in the 1860's called Rheinberg), as well as calibration experiments and measurements. The teachers were incredibly impressed with the ability to transfer the image from the microscope to the video monitor. (Actually, they all instantly reverted from being mature, in-control adults to enthusiastic 7 year-olds! ... and I mean that in the most complimentary sense).
As for equipment: For the simplest connection, just the camera and a video monitor. the cameras typically come with some sort of clamping device. Just remove the eyepiece and replace with the camera and adapter.
For the next level up, you can get an analog-to-digital (ADC) video capture card that goes into the computer. The camera then hooks into the back of the card and the card, in conjunction with some special software, captures the images and makes them available for storage and manipulation in the computer.
Although I've not researched the market lately, I think that all of the inexpensive cameras (like the Flexcam), are analog. They require both the electronic interface and software to communicate with a computer and produce digital images.
Once you have the images in digital format, you can use your computer and digital projector to project the still images in the classroom. Alternatively, using the simple set up described above, you can project live moving objects, like pond critters and crystal growth.
I think you will find a web search and the camera manufacturers a wealth of info, but contact me off-line if you have further questions.
Good hunting! .... and have fun!!!
Best regards, Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 04:09 PM 1/4/2005, jfrancis-at-powell.k12.ky.us wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 12:02:31 2005
Jennifer; A number of years ago, Intel and Mattel teamed up to produce a very inexpensive microscope that interfaced to a computer. Unfortunately, they ceased production, although they can be found on Ebay. (I just checked-there are 16 on Ebay at this moment, with the highest price being $40) I regret that I never bought one. Everyone I know who bought one for their kids was delighted. BTW, I do work for Intel, but have I no connection at all to this microscope, and as I said, it is out of production.
John Mardinly
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:jfrancis-at-powell.k12.ky.us] Sent: Tuesday, January 04, 2005 2:10 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jfrancis-at-powell.k12.ky.us) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, January 4, 2005 at 11:37:24 ------------------------------------------------------------------------ ---
Email: jfrancis-at-powell.k12.ky.us Name: Jennifer Francis
Organization: Powell County Schools
Education: 6-8th Grade Middle School
Location: Stanton,KY, USA
Question: I am formerly an elementary school teacher, now working with middle school students. I would like to invest in a microscope that we can view on a tv or computer monitor. Can you tell me what equipment I might need to do that? I am not sure what the differences are between a digital microscope, a flex cam, etc. The digital microscopes seem to be less expensive, and I wondering if I connect that to a computer/laptop if I could use a digital projector (already own) to project from there and if that is better than something like the flex cam? Would you have any ideas? Thanks!
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Jennifer -
The answer to your question has to start with the politics of education. Digital microscopes with a computer feed qualify for "technology" funds, which makes them easier to purchase in many school districts. Your classroom computer must be new enough to have a USB port. Yes, the projector will work fine. Flex cams are available with either analog (TV) or digital (PC) feed, and are a good choice if you already have a microscope; they start at around $300. An advantage of the flex cam approach is that it will work with both compound and dissecting scopes, and many of the things that middle school students want to observe (bugs, rocks, flowers, etc.) are best observed with a dissecting scope. A basic monocular dissecting scope costs $75.
That said, I urge you to think about what your students will see with that digital projector; another image on a monitor, passively. From the Project MICRO point of view, I'd like THEM to be the observers! At the very least, buy a dozen or so "flashlight" style 30x microscopes (~$10 each) as a supplement, so that each student can learn to observe. You may find the advice on buying school microscopes that is part of the MICRO web page helpful (URL below). -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 15:17:28 2005
This microscope is still available, now from Digital Blue: http://www.playdigitalblue.com/products/ There's the old QX3, and a new "improved" QX5. The quote marks are because the QX5 no longer supports TWAIN, so it can't be run through, say, Photoshop, like the QX3 can be -- the 5 only runs with the proprietary software. Pity, that.
Phil
} Jennifer; } A number of years ago, Intel and Mattel teamed up to produce a } very inexpensive microscope that interfaced to a computer. } Unfortunately, they ceased production, although they can be found on } Ebay. (I just checked-there are 16 on Ebay at this moment, with the } highest price being $40) I regret that I never bought one. Everyone I } know who bought one for their kids was delighted. BTW, I do work for } Intel, but have I no connection at all to this microscope, and as I } said, it is out of production. } } John Mardinly } } -----Original Message----- } } From: by way of Ask-A-Microscopist [mailto:jfrancis-at-powell.k12.ky.us] } Sent: Tuesday, January 04, 2005 2:10 PM } To: microscopy-at-microscopy.com } Subject: [Microscopy] AskAMicroscopist: microscope that we can view on a } tv or computer monitor } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 16:05:21 2005
I second this recommendation for the Intel QX3, and if you are a mac user there is a great freeware app called Mixscope that will capture jpegs, tiffs, and timelapse quicktime movies from the QX3 on OS X.
http://homepage.mac.com/aireck/qx3/
best regards, kevin
----- Original Message ----- } From: "Mardinly, John" {john.mardinly-at-intel.com}
I have one of these Intel 'scopes and am very unhappy with it. We never use it. A cheap video camera and a monitor is so much better.
Here's a really simple and cheap approach (certainly easier than the BX61 we're trying to set up!): http://www.aecom.yu.edu/aif/gallery/cheapscope/index.htm which also shot the pictures, but with a 3 megapixel camera, at http://www.flushart.com/gla/livinglake/index.htm
Regardless, based on my experience having done a project with two second grade classes in September, the kids really need to look into the barrel of the microscope themselves to get excited.
-Michael C.
} Jennifer; } A number of years ago, Intel and Mattel teamed up to produce a } very inexpensive microscope that interfaced to a computer. } Unfortunately, they ceased production, although they can be found on } Ebay. (I just checked-there are 16 on Ebay at this moment, with the } highest price being $40) I regret that I never bought one. Everyone I } know who bought one for their kids was delighted. BTW, I do work for } Intel, but have I no connection at all to this microscope, and as I } said, it is out of production. } } John Mardinly
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ **This electronic transmission contains information that is privileged.**
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 16:44:01 2005
} Optical Elements Corporation (OPELCO) has current openings for a Light Microscope and a Confocal Microscope Sales Specialist in the Washington, DC region. If interested, please click on the following link or contact me directly. } http://www.opelco.com/employmentcontact.htm } } Best Regards, } Cliff Glier } COO } OPELCO } 105 Executive Drive Suite 100 } Dulles, VA 20166 } 703.471.0080 x230 } 703.904.9432 (fax) } cglier-at-opelco.com } www.opelco.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 16:44:22 2005
I need some expert advice so I can help a user in my lab. He wants to see some nannoparticles he has synthesized by the following procedure:
} The sample is CdTe, a highly fluorescent NP with a shell of either } thioglycolic acid or 2-mercaptoethylamine. In theory they should be about } 2-5nm in diameter, but they are synthesized in aqueous solution, and in } order to properly seperate the particles I perform a ligand exchange and } redissolve into organic solvents(ie toluene).
So, he shows up and I put his solution onto a carbon coated formvar grid. I look around. I don't see much, some junk, but nothing like nannoparticles. He is disappointed.
I am scratching my head. Is there something there and I can't see it? Would I see it if it were there?
Maybe you have some ideas?
Would raw CdTe particles at 2 nm size have enough contrast to show up?
Could the solution be so concentrated that it looks like a solid field rather than separate particles?
The solution he gave me didn't really dry on the grid like I thought it would. How fast does toluene evaporate and could it mangle the formvar?
Any other helpful hints to get some results?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 5 21:26:44 2005
} I need some expert advice so I can help a user in my lab. He wants to } see } some nannoparticles he has synthesized by the following procedure: } } } The sample is CdTe, a highly fluorescent NP with a shell of either } } thioglycolic acid or 2-mercaptoethylamine. In theory they should be } } about } } 2-5nm in diameter, but they are synthesized in aqueous solution, and } } in } } order to properly seperate the particles I perform a ligand exchange } } and } } redissolve into organic solvents(ie toluene). } } So, he shows up and I put his solution onto a carbon coated formvar } grid. I } look around. I don't see much, some junk, but nothing like } nannoparticles. } He is disappointed. } } I am scratching my head. Is there something there and I can't see it? } Would } I see it if it were there? } } Maybe you have some ideas? } } Would raw CdTe particles at 2 nm size have enough contrast to show up? } } Could the solution be so concentrated that it looks like a solid field } rather than separate particles? } } The solution he gave me didn't really dry on the grid like I thought it } would. How fast does toluene evaporate and could it mangle the formvar? } } Any other helpful hints to get some results? } Dear Jon, I have had a few experiences looking at quantum dots and zeolite precursors for some of the materials scientists and chemical engineers here, and I may be able to answer some of your questions. First of all, these objects are small and inherently hard to see, so it would be easy to go to a relatively high mag, scan across a grid square or two and not see much. I used cryofixation and looked at frozen-hydrated material, which is generally lower contrast than particles on thin carbon; however, the uniformity of the background could be better for ice than for a carbon coat, and this would render the particles more visible. CdTe certainly has more contrast than what I was looking at, so that is not the problem. It is very unlikely that the material is too concentrated to see. Especially since the evaporation of the toluene was not as expected, the particles are much more likely to aggregate than to form a uniform layer. If you have an oven or even a warm room, you might try drying the toluene at a somewhat elevated temperature. If the toluene mangled the formvar, I'd expect to see prominent, irregular features (as happens when one gets poor formvar removal using chloroform). If the particles as prepared are well-dispersed in the aqueous solution, and if you have access to cryopreparation equipment, you could try looking at plunge-frozen specimens. Measuring the fluorescence of the solution should allow you to calculate the number of particles per microliter, so you could determine roughly how many particles you would expect to see in a field at the magnification you use. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 06:49:39 2005
Does anyone have a recommendation for what make/model of sputter coater to buy, if one was going to buy one? Currently our SEM stub coating needs are met by an ancient Edwards evaporator (which is still working, by the way). I know there's a number of other things one can do with an evaporator that can't really be done with a sputter coater, but we don't do those things - we really just need something to coat the occasional SEM stub, in case the old Edwards unit finally buys the farm. So if I was to have a small amount of capital dumped on me for such a purchase, how much would I have to spend for a reasonably reliable little sputter coater, and which one(s) should I consider?
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 08:29:24 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sbledsoe-at-iupui.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 6, 2005 at 07:23:28 ---------------------------------------------------------------------------
Email: sbledsoe-at-iupui.edu Name: Sharon B Bledsoe
Organization: Indiana University, School of Medicine
I know that individual 5nm gold spheres start to become difficult to see in a real sample on carbon/formvar grids. This is not so much size as the density you will get against a relatively thick background coating (} 50nm). Cd and Te are both lighter than Au and if the particles are not too symmetrical then I would have thought that they would be practically invisible.
It sounds like one of those classical e.m. problems where the resolution of the microscope is not the issue but the resolution and contrast of the sample may be. I suppose you could play around with the voltage and aperture size to enhance contrast a bit, as well. Someone has already mentioned shadowing but I wonder if a simple negative stain might help - I really have no idea. But I'd be interested to hear how you get on.
It would certainly be worth trying with the bare particles if you can.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: jmkrupp-at-cats.ucsc.edu (Jon Krupp)
Hi all,
I am looking for a protocol for epoxy processing of 8-10um previously sectioned cryoembedded tissue. I last did this about 10 years ago and can not locate my notes. I recall stepping the slide through the processing solutions with shortened times in a covered coplin jar until the final infiltration steps when the plastic was pipetted directly onto the slide's surface over the section and placed under vacuum. I also remember embedding by inverting the slide over an over filled beam capsule, polymerizing, then separating the two by using LN2 to pop the slide away from the plastic block.
If you have recommendations for the duration of the processing steps, I'd appreciate hearing them.
Thank you, Mary Ellen Pease
Mary Ellen Pease, M.S. Laboratory Manager Glaucoma Research Lab & Microscopy and Imaging Core Facility Wilmer Eye Institute, Johns Hopkins Hospital 175 Woods Research 600 N. Wolfe Street Baltimore, MD 21287 410-955-3337 (phone/voicemail) 443-287-2711 (fax) mpease-at-jhmi.edu
WARNING: Email sent over the Internet is not secure. Information sent by email may not remain confidential. DISCLAIMER: This email is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you receive this email by mistake, notify the sender and destroy the email.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 09:54:29 2005
Frank, Although I may not be able to make an ultimate recommendation for purchase I would suggest you avoid Cressington. We have their 208HR model sputter coater with mtm20 thickness controller and have been generally pleased with the operation of the unit. We use either Pt/Pd or Cr targets to coat polymeric materials. What has been especially unsatisfactory about the unit is its 'voracious appetite' for targets. Typically our Pt/Pd targets (57mm dia. x 0.1mm thick) last 2 to 4 months and our usage is not heavy. The unit's power is concentrated in a central ring of about 20mm and when the targets fail a small crescent-shaped hole remains where the Pt/Pd has been etched away. I made some measurements of the actual consumption of Pt/Pd upon failure and found it to be only 8 to 12%. When you are spending approx. $500 US per target and you are left with about $450 US of waste Pt/Pd you can see that it is a very inefficient use of your money. Apparently Cressington have been developing a smaller magnetron head which would accept a smaller diameter target and hence reduce the amount of waste target material. One other dislike of the unit in its present configuration is the inability to maintain vacuum in the chamber once power is switched off. I am unaware if their newer models reflect improvements in these two areas.
Good luck in your search.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Thursday, January 06, 2005 7:55 AM To: microscopy-at-microscopy.com
Listers -
Does anyone have a recommendation for what make/model of sputter coater to buy, if one was going to buy one? Currently our SEM stub coating needs are met by an ancient Edwards evaporator (which is still working, by the way). I know there's a number of other things one can do with an evaporator that can't really be done with a sputter coater, but we don't do those things - we really just need something to coat the occasional SEM stub, in case the old Edwards unit finally buys the farm.
So if I was to have a small amount of capital dumped on me for such a purchase, how much would I have to spend for a reasonably reliable little sputter coater, and which one(s) should I consider?
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 11:12:32 2005
----- Original Message ----- } From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Karen Bovard" {kbovard-at-creighton.edu} Sent: Thursday, January 06, 2005 9:15 AM
Dear Paul, One solution to your problem is to contact Abe Dayani (tel. 702-368-0579) at Refining Systems Inc. (http://www.refiningsystems.com/) for new sputtering targets and a credit on the unused materials in your spent targets. He prices his targets on the precious metal content and they are usually about half the price of the targets from the manufacturer. No financial interest, just a satisfied customer. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Gerroir, Paul" {paul.gerroir-at-xrcc.xeroxlabs.com} To: "Thomas, Frank" {FThomas-at-nrcan.gc.ca} ; {microscopy-at-microscopy.com} Sent: Thursday, January 06, 2005 7:57 AM
Dear Frank, I received a used Denton DeskII when I bought a used SEM and it has been a good, solid performer. It is not fancy, but it is inexpensive, easy to service and reliable. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 12:00:44 2005
SEM or TEM? For either, we just deposit the particles on Formvar coated grids, with or without C, directly from the water solution. Just as long as their are no salts or the like to precipitate. This works with colloidal particles down to 3 - 5 nm, and composed of Au, Pt, Pd, Ag, Fe, and combinations and alloys of these. The particles separate fine without doing anything else. This is also true for particles conjugated to proteins like antibodies. I did try to look at some home-made Cd/Se quantum dots that had been synthesized in toluene (and a couple of other organic solvents, but I forget which), and it was a no-go. The toluene attacked and either dissolved or "wadded up" the formvar film. This may be what happened to your samples.
Phil
} Hi: } } I need some expert advice so I can help a user in my lab. He wants to see } some nannoparticles he has synthesized by the following procedure: } } } The sample is CdTe, a highly fluorescent NP with a shell of either } } thioglycolic acid or 2-mercaptoethylamine. In theory they should be about } } 2-5nm in diameter, but they are synthesized in aqueous solution, and in } } order to properly seperate the particles I perform a ligand exchange and } } redissolve into organic solvents(ie toluene). } } So, he shows up and I put his solution onto a carbon coated formvar grid. I } look around. I don't see much, some junk, but nothing like nannoparticles. } He is disappointed. } } I am scratching my head. Is there something there and I can't see it? Would } I see it if it were there? } } Maybe you have some ideas? } } Would raw CdTe particles at 2 nm size have enough contrast to show up? } } Could the solution be so concentrated that it looks like a solid field } rather than separate particles? } } The solution he gave me didn't really dry on the grid like I thought it } would. How fast does toluene evaporate and could it mangle the formvar? } } Any other helpful hints to get some results? } } Thanks } } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 14:04:49 2005
Hi all, I have a user who wants to stain lignin in JB-4 resin-embedded corn stalks. He tried Phloroglucinol which works fine in paraffin embedded samples since the paraffin is removed. However, this stain requires an HCl treatment to reveal the desired color. The acid treatment interacts adversely with the JB-4 resin.
The resin embedding gives much better resolution than the paraffin so it is not an option to go back to the paraffin-embedded tissue.
Does anyone have an alternative stain that will work with this resin?
Thanks in advance, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 14:42:37 2005
I am currently using the 4pi Revolution system to capture digital images on my JEOL 840 (and two other SEMs as well). You can find the information on the system at:
http://www.4pi.com/
I have been very pleased with the system for a number of years and also would point out that 4pi offers integrated EDS or WDS analysis with the imaging system.
Scott Miller (No financial interest in 4pi, just a very satisfied customer)
F. Scott Miller, Ph.D. Advanced Materials Characterization Lab University of Missouri - Rolla 223 McNutt Hall Rolla, MO 65409 USA fax: 573 341 6934 voice: 573 341 4727
} ---------- } From: Karen Bovard } Sent: Thursday, January 6, 2005 8:44 AM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Image grabbing systems for SEM } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I have a JEOL 840A SEM and am interested in upgrading it to digital } photograpy capabilities. } } I am aware of the Orion, SIS ADDA II, and the JEOL Orion systems. } } Are there any different options (preferably cheaper) to consider? } } Karen Bovard } EM Lab } Pathology } Creighton University } Omaha, NE } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 14:54:02 2005
Imapro QCRZi 35mm Film Recorder. Comes with 35mm module & instruction manuals. Utilizes GPIB interface. Has cables, software and instruction manuals and PCI card from National Instruments. Running on Mac OS 9.2, the RIP software is Graphx RasterPlus includes all manuals. Includes Power Mac 8500 computer and 19" Hitachi monitor. In excellent working condition $500 USD or best offer. Please contact Ian Craig directly at: Ian Craig Media Specialist Faculty of Science The University of Western Ontario 591-661-2111 ext.86778 icraig-at-uwo.ca
Richard Harris Laboratory Supervisor Department of Biology University of Western Ontario London Ontario CANADA N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 6 15:33:05 2005
I think formvar support film is not good for such type of experiment. You need thin carbon film to reduce scatter from background and enhance signal-to-noise ratio. I had difficulties to see nanogold particles (about 2 nm in diameter) in bright but dark field. In dark field on 1.8 nm carbon they are perfectly visible. Your particles has lover density, so it would be even trickier to see them than gold. Again, sometime it's easier to see the particles on the film rather on the screen (so you focus on some junk in hope to have your object nearby). Anyway, I think it's not such easy to see nanosize object on the any support film. Negative staining would just complicate the situation because UA staining for instance generated approximately 0.8 nm granularity of background. I don't think you could resolve 2 nm object well with 0.8 nm probe at least at the negative staining condition. Sergey
At 07:12 AM 1/6/2005, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
I bought Epo-tek Epoxy 353ND since I heard Gatan G1 and 353ND are the same. However, no instruction came with the epoxy. On the website I found that Cure Time: 1 min.at 150°C or 5 min. at 100°C. For embedding materials, what temperature and time is the best? Please advise.
Thank you, Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 7 03:09:05 2005
.. I have no commercial interests or benefits with this product, only detailed knowledge about the functionality (from my jobs in former times).
Best regards
Frank
Karen Bovard wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Jan 7 06:01:47 2005
Hello, Its quit difficult to view the 2nm particles, as such being very tiny and thin they will have a very little contrast and it further depend on the atomic weight.
We have successfully viewd 2-3nm CdS particles on formar film. Yes you have to play with the aperature and accelarting voltages and spot size. According to my personal experience, though at 200kv you will get the better resolution but for such kind I maily use the accelarating voltage of 120kv which gives better contrast. Moreover the alignment should be perfect. We use to align the Gun everytime and correct the astigmata for such kind of samles moreover one need absolute concentration and patience for such kind of samples. Many times I too use to get frusated with such kind of samples but then slowly i started getting the good results. Pl do not hesistate to contact if u need any further information Goodluck
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I think formvar support film is not good for such } type of experiment. You } need thin carbon film to reduce scatter from } background and enhance } signal-to-noise ratio. I had difficulties to see } nanogold particles (about } 2 nm in diameter) in bright but dark field. In dark } field on 1.8 nm carbon } they are perfectly visible. Your particles has } lover density, so it would } be even trickier to see them than gold. Again, } sometime it's easier to see } the particles on the film rather on the screen (so } you focus on some junk } in hope to have your object nearby). Anyway, I think } it's not such easy to } see nanosize object on the any support film. } Negative staining would just } complicate the situation because UA staining for } instance generated } approximately 0.8 nm granularity of background. I } don't think you could } resolve 2 nm object well with 0.8 nm probe at least } at the negative } staining condition. Sergey } } At 07:12 AM 1/6/2005, you wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Jonathon } } } } I know that individual 5nm gold spheres start to } become difficult to } } see in a real sample on carbon/formvar grids. This } is not so much size } } as the density you will get against a relatively } thick background } } coating (} 50nm). Cd and Te are both lighter than Au } and if the } } particles are not too symmetrical then I would have } thought that they } } would be practically invisible. } } } } It sounds like one of those classical e.m. problems } where the } } resolution of the microscope is not the issue but } the resolution and } } contrast of the sample may be. I suppose you could } play around with the } } voltage and aperture size to enhance contrast a } bit, as well. Someone } } has already mentioned shadowing but I wonder if a } simple negative stain } } might help - I really have no idea. But I'd be } interested to hear how } } you get on. } } } } It would certainly be worth trying with the bare } particles if you can. } } } } Malcolm } } } } Malcolm Haswell } } e.m. unit } } School of Health, Natural and Social Sciences } } University of Sunderland } } Tyne & Wear } } SR1 3SD } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } ----- Original Message ----- } } } From: jmkrupp-at-cats.ucsc.edu (Jon Krupp) } } Date: Wednesday, January 5, 2005 10:47 pm } } Subject: [Microscopy] Looking for CdTe } nannoparticles } } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line } Help } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hi: } } } } } } I need some expert advice so I can help a user } in my lab. He wants } } } to see } } } some nannoparticles he has synthesized by the } following procedure: } } } } } } } The sample is CdTe, a highly fluorescent NP } with a shell of either } } } } thioglycolic acid or 2-mercaptoethylamine. In } theory they should } } } be about } } } } 2-5nm in diameter, but they are synthesized in } aqueous solution, } } } and in } } } } order to properly seperate the particles I } perform a ligand } } } exchange and } } } } redissolve into organic solvents(ie toluene). } } } } } } So, he shows up and I put his solution onto a } carbon coated } } } formvar grid. I } } } look around. I don't see much, some junk, but } nothing like } } } nannoparticles.He is disappointed. } } } } } } I am scratching my head. Is there something } there and I can't see } } } it? Would } } } I see it if it were there? } } } } } } Maybe you have some ideas? } } } } } } Would raw CdTe particles at 2 nm size have } enough contrast to show up? } } } } } } Could the solution be so concentrated that it } looks like a solid field } } } rather than separate particles? } } } } } } The solution he gave me didn't really dry on the } grid like I } } } thought it } } } would. How fast does toluene evaporate and could } it mangle the } } } formvar? } } } Any other helpful hints to get some results? } } } } } } Thanks } } } } } } } } } Jonathan Krupp } } } Microscopy & Imaging Lab } } } University of California } } } Santa Cruz, CA 95064 } } } (831) 459-2477 } } } jmkrupp-at-cats.ucsc.edu } } } } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-080 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } } }
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From MicroscopyL-request-at-ns.microscopy.com Fri Jan 7 12:00:25 2005
The lower the temp, the longer the time. It will not cure at room temperature. However, if you go too hot, the stuff fumes a bit, so be careful. I use a hot plate at 120C, but I can't tell you the time since I use a small vise and it takes time for the vise to get up to temperature. There is a color change with this epoxy. I use a small dollop on a piece of Teflon that I can take off to tell me when it is done. The color goes from a clearish yellow to a deep brownish red. I would test it with your process. Use a sharp Exacta blade or a razor blade and poke it when you think that it looks done. When it is done, it will be hard.
-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: hkonishi-at-indiana.edu [mailto:hkonishi-at-indiana.edu] Sent: Thursday, January 06, 2005 4:57 PM To: message to: MSA list
Hello
I bought Epo-tek Epoxy 353ND since I heard Gatan G1 and 353ND are the same. However, no instruction came with the epoxy. On the website I found that Cure Time: 1 min.at 150°C or 5 min. at 100°C. For embedding materials, what temperature and time is the best? Please advise.
Thank you, Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 7 12:24:18 2005
I worked with a client earlier this week using SEM/EDS to characterize some WW-II medals. The client wanted to determine the nature of authentic medals so that forgeries might be easily detected.
I thought the session to be fairly straightforward. We looked at both the paint and the metals involved and were able to clearly show a few things that had never been seen before. That is, it appears that nobody had ever looked at such items by SEM and EDS before. That seems strange to me, but I guess there are still a lot of things that have not been examined.
Well, now I have been contacted by another collector from the NY area who is wondering if labs are available there to do similar work. I have forwarded his request below so that interested parties may contact him directly. I suppose you may also contact me for more details about the work.
Warren
} } From popserve Wed Jan 5 13:39:02 2005 } Date: Wed, 5 Jan 2005 11:32:21 -0800 (PST) } From: Marc Garlasco {marc-at-garlasco.com} } Subject: [Microscopy] Tom Hansen } To: wesaia-at-iastate.edu } X-PMX-Version: 4.7.0.111621, Antispam-Engine: 2.0.0.0, Antispam-Data: } 2005.1.5.1 } X-Perlmx-Spam: Gauge=IIIIIII, Probability=7%, Report='__CT 0, } __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_VERSION 0, __SANE_MSGID 0' } } Mr. Straszheim, } } I am a friend of Tom's and I am amazed at the } cutting-edge work you guys did on the crosses } yesterday! I would like to enlarge the pool of data } by getting my crosses analyzed here in the NY area. } Can you suggest good departments to contact? } } Regards, } Marc Garlasco
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Here is the January 2005 Microscopy Today table of contents. I will close the subscription list for this issue on Tuesday 11 January 2005.
THIS ISSUE CONTAINS THE MT SALARY SURVEY RESULTS
Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Unfolding and Folding Proteins Stephen W. Carmichael, Mayo Clinic
Spiral Powder Overlays P. Fraundorf and Shuhan Lin
TEM Morphometry Reveals Membrane Deficits in Parietal Cells Lacking Specific Ion Transporters Miller ML, Gawenis LR, Andringa A, Shull GE
Color Matching to Ink Jet Printers from a Computer Screen, Part 2 Jerry Sedgewick
Electron Tomography in the Study of Bacterial Structure and Function Kenneth H. Downing,* Haixin Sui,* Luis R. Comolli* and Hoi-Ying Holman
Having Your Cake and Eating It Too: A Procedure for Obtaining Plan View and Cross Section TEM Images from the Same Site R.B. Irwin, A. Anciso, P.J. Jones, and C. Patton
Confocal Scanning Laser Holography: A Tool for Non-Invasive Internal Measurement RA McLeod, P Jacquemin, S Lai, RA Herring
Dehydration and Rehydration Issues in Biological Tissue Processing for Electron Microscopy Christian T. K.-H. Stadtländer
Microscopy Today 2004 Salary Survey Results Ron Anderson and Barbara Foster
Funding Opportunities for Acquiring Major Equipment from Federal Granting Agencies M&M 2004 Core Facility Management – Part I: NIH Organizer: Debby Sherman
Specimen Capsules For Critical-Point Drying Sol Sepsenwol
Industry News
NetNotes
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 7 12:53:54 2005
Jon- We have successfully imaged 1 nm particles under even more adverse conditions. That is, in 70-nm thick sections of epoxy resin.
One has to collect two digital images and subtract them. The one is run through a 3x3 kernel to smooth it, and the 1-nm gold image drops out of that one.
I don't know whether you have digital image collection and processing, but if you have, this would be an easy way to solve your problem.
} X-Authentication-Warning: ns.microscopy.com: mail set sender to } MicroscopyL-request-at-ns.microscopy.com using -f } X-Sender: jmkrupp-at-cruzmail.ucsc.edu } Date: Wed, 5 Jan 2005 14:47:17 -0800 } To: microscopy-at-microscopy.com } From: jmkrupp-at-cats.ucsc.edu (Jon Krupp) } Subject: [Microscopy] Looking for CdTe nannoparticles } X-UCSC-CATS-MailScanner: Found to be clean } X-UCSC-CATS-MailScanner-SpamCheck: } X-MASF: 0.00% } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu http://www.bgsu.edu/departments/biology/facilities/MnM ___________________________________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 7 15:37:29 2005
As my experience shows it may be additional benefits in terms of strength is you can increase a little bit humidity during the curing. Water vapour, for example, placing an open bottle filled with water at some place closer to your bonding. Hope it will help.
Victoria
----- Original Message ----- } From: "Walck, Scott D." {walck-at-ppg.com} To: {hkonishi-at-indiana.edu} Cc: "MicroscopyListserver (E-mail)" {Microscopy-at-microscopy.com} Sent: Friday, January 07, 2005 9:44 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rebecca.burgin-at-onsemi.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 7, 2005 at 11:34:19 ---------------------------------------------------------------------------
Question: We are in the market for a used Hitachi S4800/S4700 SEM (or comparable). Please contact me at 602-244-5775 (phone) or rebecca.burgin-at-onsemi.com (email)if you have one available or have any leads. Thanks in advance.
A while back I tried to locate a 21X BF/DF objective for an antique Bausch & Lomb Research Metallograph that I am bringing back to life. So far, the solution hasn't come into coincidence with my limited budget. However, I am getting "close enough" by adapting a short-barrel Carl Zeiss 21X objective which gives excellent images on the Kodak MDS 100 digital camera that I've adapted to the metallograph.
However, the Zeiss objective was designed for a tube length of 190 mm, but the metallograph uses a tube length of 215 mm. The images are good enough that I really ought to accept the present situation. However, I am nevertheless trying to do it right by adding a compensating lens in the light path. However, I have not yet found out how to do the math. I understand that a negative lens is needed to stretch the image distance from 190mm to 215mm. I even have the Bausch & Lomb compensating lens that comes with their vertical-illuminator attachment for transmitted-light microscopes, but that lens is designed to shift the image distance by 55mm, i.e., the extra optical path length introduced by the vertical illuminator.
Can anyone steer me to a suitable on-line or library reference that shows how to do the calculation ?
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 8 19:38:41 2005
On 8 Jan 2005, at 9:03, George Langford, Sc.D. wrote: } } Hello Microscopists ! } } A while back I tried to locate a 21X BF/DF objective for an antique Bausch & } Lomb Research Metallograph that I am bringing back to life. So far, the } solution hasn't come into coincidence with my limited budget. However, I am } getting "close enough" by adapting a short-barrel Carl Zeiss 21X objective } which gives excellent images on the Kodak MDS 100 digital camera that I've } adapted to the metallograph. } } However, the Zeiss objective was designed for a tube length of 190 mm, but the } metallograph uses a tube length of 215 mm. The images are good enough that I } really ought to accept the present situation. However, I am nevertheless } trying to do it right by adding a compensating lens in the light path. } However, I have not yet found out how to do the math. I understand that a } negative lens is needed to stretch the image distance from 190mm to 215mm. I } even have the Bausch & Lomb compensating lens that comes with their } vertical-illuminator attachment for transmitted-light microscopes, but that } lens is designed to shift the image distance by 55mm, i.e., the extra optical } path length introduced by the vertical illuminator. } } Can anyone steer me to a suitable on-line or library reference that shows } how to do the calculation ?
Sorry, can't help you with the math, but the following link will show the tolerance of dry objectives to changes in tube length based on the n.a of the lens.
or go to http://groups.yahoo.com/group/Microscope, on the left side of the page, click on Files, scroll down to folder named "Microscope Theory and Figures" click and then click on Tube length deviation.jpg
Unless your objective has an n.a. } 0.5 you won't introduce any measurable distortion in the image. You might have to piece the long link back together in your browser.
Bob Sunley
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 03:56:31 2005
Karen Bovard wrote: ======================================================================= I have a JEOL 840A SEM and am interested in upgrading it to digital photograpy capabilities.
I am aware of the Orion, SIS ADDA II, and the JEOL Orion systems.
Are there any different options (preferably cheaper) to consider?
Karen Bovard EM Lab Pathology Creighton University Omaha, NE ========================================================================== The "cheapest" option, which was offered by SPI Supplies for many years was Spectrum Mono (previously known in some parts of the world as Image Slave). However it is no longer being offered by the manufacturer.
We are now offering the ORION™ Digital Image System for SEMs, or in simple words, the Orion "frame grabber". See URL http://www.2spi. com/catalog/instruments/ORION_Digital_Image_Acquisition_System.html Various optional modules are also available to extanding the software's capabilities.
It is a passive image capture system, is easy to operate and easy to install We use it all the time in our own laboratory and have found it to be quite easy to learn to use as well. An active image capture system it is not, but then again, there are many who do not need the benefits of an active capture system (which is much more expensive as well).
This system should not be confused with the JEOL Orion system. It is unfortunate that two firms are using the same trade name since it is bound to cause confusion in the marketplace. These are most certainly not the same product.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 08:08:16 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Question: Hello, does anyone out there have experience in SEM on lubricates or grease? I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one?
} From their Web site: 'QuantomiX develops and commercializes breakthrough solutions based on its proprietary wetSEM™ technology which enables direct scanning electron microscopy (SEM) of wet samples.'
I have no experience of using the holders or any connection with the company. I've just seen them advertising.
Ron
On Mon, 10 Jan 2005 08:16:53 -0600 by way of MicroscopyListserver {diller-at-stefan-diller.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } 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 (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 } --------------------------------------------------------------------------- } } Email: diller-at-stefan-diller.com } Name: Stefan Diller } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hello, } does anyone out there have experience in SEM on lubricates or grease? } I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... } My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one? } } Thanks for your help. } Stefan Diller } } --------------------------------------------------------------------------- } }
---------------------- 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 http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 11:50:28 2005
I think you may need to do frozen imaging or cryoSEM. It might work doing a freeze-fracture-etch and then image with a high resolution coating. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha 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
On Mon, 10 Jan 2005, by way of MicroscopyListserver wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } 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 (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 } --------------------------------------------------------------------------- } } Email: diller-at-stefan-diller.com } Name: Stefan Diller } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hello, } does anyone out there have experience in SEM on lubricates or grease? } I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... } My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one? } } Thanks for your help. } Stefan Diller } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 12:18:15 2005
I have experience looking at greases and lubricants in the SEM (JEOL JSM-5800LV). One such request had nanoparticles in the grease. Unfortunately, my results aren't promising.
I don't really see how a lubricant can be imaged without using an environmental chamber SEM. I had limited success when the lubricant thinned out enough to where I can begin to see the nanoparticles. It also helped that the particles were tungsten-based for backscatter image contrast. Resolution was poor. EDS analysis was somewhat doable.
Can you perhaps separate or concentrate the particles by using solvent and centrifuge techniques before imaging? This may help.
Stu Smalinskas, P.E. SKF USA Plymouth, Michigan (734) 414-6862
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Stefan wrote:
Question: Hello, does anyone out there have experience in SEM on lubricates or grease? I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one?
Thanks for your help. Stefan Diller
Email: diller-at-stefan-diller.com Name: Stefan Diller
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 12:41:44 2005
In do have a cheap and cheerful way to overcome your problem. One feature however may make my explanation null and void! You need a microscope that has a manifold directly pumped by a diffusion or turbo pump, not a manifold that bleeds the specimen vacuum to the gun? You also need a backscattered electron detector. In this procedure we are taking advantage of a poor vacuum bleeding away surface charge and reducing media evaporation from the specimen.
If you have a manifold system and a backscattered detector the following works very well for up to 20 minute working periods. Take a rubber bung/stopper, that will fit into the pumping line at the rear of the specimen chamber, freeze it in liquid nitrogen and drill a 0.5mm hole in the bung/stopper. Prepare your specimen and place it in to the microscope at the same time fitting the bung/stopper in to the pumping line. Pump down and switch off the SE detector bringing the BSE detector into play. Use the BSE detector to observe the specimen in the "poor vacuum" environment that you have created. I have used this technique many times, the only drawback is that the many manufacturers took us away from a decent vacuum system when they decided to pump the column through the specimen chamber, rather than pumping the microscope through a manifold.
So users of older microscopes have the cheapest possible "VP System", but those caught in the middle era in SEM development miss out on this one I am afraid!
Good luck
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide www.emcourses.com tel +44 1280 816512 fax +44 1280 814007
PS I probably should not point out that we once used this system to watch paint dry!
----- Original Message ----- } From: "by way of MicroscopyListserver" {diller-at-stefan-diller.com} To: {microscopy-at-microscopy.com} Sent: Monday, January 10, 2005 2:16 PM
One of my colleagues many years ago at Lockheed, George Hopple, had excellent success critical point drying greases. He was then able to image the thickener and the oil, and the quite striking differences in the greases were successfully tied to bearing failures in gyroscopes. I don't have any contact information for George at this time, as he left Lockheed over a decade ago, but maybe somebody out there can find him.
John Mardinly Intel
-----Original Message----- } From: by way of MicroscopyListserver [mailto:diller-at-stefan-diller.com] Sent: Monday, January 10, 2005 6:17 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 ------------------------------------------------------------------------ ---
Email: diller-at-stefan-diller.com Name: Stefan Diller
Question: Hello, does anyone out there have experience in SEM on lubricates or grease? I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one?
Hello, I was reading over some things about diatoms and wanted to get a good light microscope image of some organisms from a biofilm/water. I was wondering what is a good way of imaging live microbes with an inverted light microscope? Just use a coverslip, slide and place it upside down on the inverted stage? I read about using modified petrie dishes too. Any advice appreciated. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha 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
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 20:22:07 2005
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, I was reading over some things about diatoms and wanted to get a good light microscope image of some organisms from a biofilm/water. I was wondering what is a good way of imaging live microbes with an inverted light microscope? Just use a coverslip, slide and place it upside down on the inverted stage? I read about using modified petrie dishes too. Any advice appreciated. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha 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
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 20:38:34 2005
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Question: Hello, does anyone out there have experience in SEM on lubricates or grease? I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one?
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think you may need to do frozen imaging or cryoSEM. It might work doing a freeze-fracture-etch and then image with a high resolution coating. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha 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
On Mon, 10 Jan 2005, by way of MicroscopyListserver wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } 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 (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 } --------------------------------------------------------------------------- } } Email: diller-at-stefan-diller.com } Name: Stefan Diller } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hello, } does anyone out there have experience in SEM on lubricates or grease? } I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... } My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one? } } Thanks for your help. } Stefan Diller } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 20:40:41 2005
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Karen Bovard wrote: ======================================================================= I have a JEOL 840A SEM and am interested in upgrading it to digital photograpy capabilities.
I am aware of the Orion, SIS ADDA II, and the JEOL Orion systems.
Are there any different options (preferably cheaper) to consider?
Karen Bovard EM Lab Pathology Creighton University Omaha, NE ========================================================================== The "cheapest" option, which was offered by SPI Supplies for many years was Spectrum Mono (previously known in some parts of the world as Image Slave). However it is no longer being offered by the manufacturer.
We are now offering the ORION™ Digital Image System for SEMs, or in simple words, the Orion "frame grabber". See URL http://www.2spi. com/catalog/instruments/ORION_Digital_Image_Acquisition_System.html Various optional modules are also available to extanding the software's capabilities.
It is a passive image capture system, is easy to operate and easy to install We use it all the time in our own laboratory and have found it to be quite easy to learn to use as well. An active image capture system it is not, but then again, there are many who do not need the benefits of an active capture system (which is much more expensive as well).
This system should not be confused with the JEOL Orion system. It is unfortunate that two firms are using the same trade name since it is bound to cause confusion in the marketplace. These are most certainly not the same product.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 20:46:08 2005
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Stefan:
I have experience looking at greases and lubricants in the SEM (JEOL JSM-5800LV). One such request had nanoparticles in the grease. Unfortunately, my results aren't promising.
I don't really see how a lubricant can be imaged without using an environmental chamber SEM. I had limited success when the lubricant thinned out enough to where I can begin to see the nanoparticles. It also helped that the particles were tungsten-based for backscatter image contrast. Resolution was poor. EDS analysis was somewhat doable.
Can you perhaps separate or concentrate the particles by using solvent and centrifuge techniques before imaging? This may help.
Stu Smalinskas, P.E. SKF USA Plymouth, Michigan (734) 414-6862
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Stefan wrote:
Question: Hello, does anyone out there have experience in SEM on lubricates or grease? I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one?
Thanks for your help. Stefan Diller
Email: diller-at-stefan-diller.com Name: Stefan Diller
__________________________________ Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. http://info.mail.yahoo.com/mail_250
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 10 20:46:35 2005
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Stefan,
Try www.quantomix.com
} From their Web site: 'QuantomiX develops and commercializes breakthrough solutions based on its proprietary wetSEM™ technology which enables direct scanning electron microscopy (SEM) of wet samples.'
I have no experience of using the holders or any connection with the company. I've just seen them advertising.
Ron
On Mon, 10 Jan 2005 08:16:53 -0600 by way of MicroscopyListserver {diller-at-stefan-diller.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } 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 (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 10, 2005 at 03:22:34 } --------------------------------------------------------------------------- } } Email: diller-at-stefan-diller.com } Name: Stefan Diller } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hello, } does anyone out there have experience in SEM on lubricates or grease? } I need to do SEM on nanoparticles in lubricates as well as various states of lubricates growing old... } My first idea is using a coolstage, but is there any possiblity (without a low vac SEM) to work around this not having one? } } Thanks for your help. } Stefan Diller } } --------------------------------------------------------------------------- } }
---------------------- 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 http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 11 08:35:17 2005
A colleague has a Philips EM400T TEM that is exhibiting odd behavior as the user mags up and down. At certain mags, the OL current goes way out, but to a reproducible figure, as follows:
At 100kV with Z-axis corrected, the OL current at true focus reads 6.66. When tracking from the upper mags toward the lower, at the transition between 130kX to 100kX, the OL current jumps to 6.44. If the user corrects back to true focus (again with an OL reading of 6.66 or 6.67), then tracking down to the next mag step causes the OL to jump again back to 6.44. This jump proceeds at every mag until the user reaches 2800X, when the OL goes to (or stays at) 6.66.
Also confusing, this sequence doesn't happen 100% of the time, and not always at every mag within the mag range described, but it does so more often than not. My guess is that a certain board that defines lens correlations in the middle-mag range has a fluky relay. Does anyone have an idea which board(s) we should be looking at to test this theory? Or does anyone have a better idea on how to address this issue?
We do have access to another EM400 that we can use for parts. Any help on how to proceed would be very helpful, and gratefully acknowledged!
Thanks all,
Ann Hein Lehman Assistant Director, Electron Microscopy Facility Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 11 16:07:05 2005
Just a piece of information to help with your decision (hopefully not to confuse you):
A PASSIVE system such as the one Mary mentions essentially takes the data generated by the SEM and digitizes them. Basically, you work with your SEM and when you see an image that you want to record, you push a button and the PC records the image. This also means, that you are limited to what the SEM can provide. If the SEM scan generator can only provide a 1000 line image, the highest resolution will be something like 1300x1000. On the other hand, passive systems can be a bit less expensive.
An ACTIVE system basically replaces the scan generator with one that is controllable by the PC. Again, you would work with your SEM normally until you see an image that you want to record. You then push a button, and the PC records an image. Different from the passive system, though, the scanning is now controlled by the PC and you can acquire the images at a higher resolution (up to 4ooox4ooo in case of our ADDA). An active system also provides control over dwell-time (noise reduction) and synchronization with 60Hz noise signals, generally resulting in better images. Furthermore, with additional hardware you can also easily acquire elemental distribution maps. An active system is usually a bit more expensive.
Disclaimer: We produce and sell the ADDA II system mentioned in Karen's original email, which is both passive and active.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Thursday, January 06, 2005 10:18 To: Microscopy
----- Original Message ----- } From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Karen Bovard" {kbovard-at-creighton.edu} Sent: Thursday, January 06, 2005 9:15 AM
According to the established TEM literature the depth of focus is:
Dfocus = Dfield x Magnification x Magnification.
With depth of field around 50 nm, which is typical imaging condition, according to the above relation the depth of focus should be about 500 meters at magnification of 100,000.
We have a CCD camera and a TV camera mounted beneath the CCD on one of our TEMs. Well, the image focused on the TV is not in perfect focus on the CCD and vice versa at magnifications in the range of 100,000 and above.
What is wrong or is there something I am missing in the Depth of focus equations?
Krassimir N. Bozhilov
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 12 02:55:16 2005
I feel that one of the 2 cameras is not well focused Find the focus of the image on fluorescent screen and compare with both cameras; you will find the one in fault Usually the CCDs cameras have a focus adjustment. Refer to your provider for the procedure The focus should be same on all cameras and fluo screen. the depth is always enough to get good focus on axial cameras.
The only cameras which usually needs to change focus are those mounted after an EELS system. best regards Mic
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According to the established TEM literature the depth of focus is:
Dfocus = Dfield x Magnification x Magnification.
With depth of field around 50 nm, which is typical imaging condition, according to the above relation the depth of focus should be about 500 meters at magnification of 100,000.
We have a CCD camera and a TV camera mounted beneath the CCD on one of our TEMs. Well, the image focused on the TV is not in perfect focus on the CCD and vice versa at magnifications in the range of 100,000 and above.
What is wrong or is there something I am missing in the Depth of focus equations?
Krassimir N. Bozhilov
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 12 09:22:05 2005
I'm looking for user feedback on low angle ion mills. Please email me directly with your experience on reliability and relative advantages of the different vendors' models.
Thanks,
Rhonda
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 12 09:39:40 2005
I'm looking at a paper that did some work with and Etec Autoscan. I'm not very familiar with the instrument, and no model number is given. Can anybody fill me a bit on the history of this SEM? What is it's vintage, W, Lab6 or FE, etc.? Does the company exist under another name? All the images and info I can find on the web look to be a 70s to 80s type instrument, but...
Gee, a "History of Microscope Manufacturers: Intrigue, Bankruptcy and Hostile Takeovers" web site would be handy...
As usual, thanks in advance!
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I am looking for a source for replacement lamps for an ancient Leitz Orthoplan microscope (ca. 1965 I think). I would appreciate any hints as to where I might purchase such. Thanks, Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 12 16:05:23 2005
I don't know the bulb number, but you might try: www.bulbdirect.com
At least they carry some Leitz bulbs...
Woody
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-ufl.edu] Sent: Wednesday, January 12, 2005 3:59 PM To: Microscopy-at-MSA.Microscopy.Com
I am looking for a source for replacement lamps for an ancient Leitz Orthoplan microscope (ca. 1965 I think). I would appreciate any hints as to where I might purchase such. Thanks, Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 12 16:51:29 2005
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Can you actually focus the image on both cameras?
If either or both of the cameras is lens coupled, then there may be a problem with the focusing of the camera itself on the scintillator.
Having said that, yes, I think that there can often be small focus differences between cameras and phosphors at different levels., especially between a wide-filed CCD mounted above the viewing chamber and a high mag CCD beneath the viewing chamber. -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 13 03:07:54 2005
I have a C. P. Goerz 3D Condenser for a microscope labeled Goerz MOM Hungary in a box marked Goerz American Optical Company, New York and a tag Universal had written and stuck on the box with a hand written number 1060 on it.
I can find no reference at all to it on the Internet. It appears to be manufactured in the Post war years from the look of the labels, box, materials and workmanship. So I suspect it was made in the 50's or 60's in Hungary and imported by American Optical.
It is fully working and in fine working condition and I would like to know any thing you have about it. Of course a copy of operating manual would be my ultimate objective.
Thanks Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 13 08:24:07 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rodel.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 13, 2005 at 07:18:54 ---------------------------------------------------------------------------
I have two LEICA Ergoplan defect review microscopes configured for 200 mm wafers. I am looking for third party service and support. The OEM charges are too high.
Both systems are running VisconNT defect review software. One is a manual load system and the second has a wafer handling robot attached. The manual load systems is having some startup errors, which might be eprom errors. The auto load systems has just been moved to my lab, and I am looking for someone to come in to install and check out the system. I can supply pictures for both tools.
Does any one know of third party support for these type of tools.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwilkinson-at-seton.org) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, January 12, 2005 at 09:45:18 ---------------------------------------------------------------------------
Question: The EM lab at Brackenridge in Austin Texas is considering a purchase of Leica specimen trimmer. I would appreciate your opinion of the valus of the trimmer. I have hear Pro's and con's and would value your experience.
Thanks to every one who responded to my query about a replacement bulb. I found one at donsbulbs.com. I am going to check out all of the other suggested sources as Don does not accept credit cards.
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 13 11:32:41 2005
Dear James, I operated an ETEC Autoscan (Manufacturer: "ETEC", model: "Autoscan") that was purchased in 1973, before I started here. It was renowned for beautiful images, very low mag capability and was a favourite SEM of its time. I believe many of David Scharf's brilliant poster images are recorded on an modified ETEC Autoscan. The Perkin-Elmer company bought ETEC up in the early '80s for their electron-beam lithography system and shut down the SEM division. There are still some running, including my old one that I sold to the military. To answer your questions: it was a conventional W filament SEM, mag. 5 to 200,000X. No FE, no VP-SEM, no computer, etc., just a good, solid SEM with top resolution about 6.0 nm. They were good for their day, but a small company that maybe couldn't keep up when more companies started to manufacture SEMs. I'm sure others know more, but no one seemed to be answering. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "James M. Ehrman" {jehrman-at-mta.ca} To: "Microscopy Listserv" {Microscopy-at-MSA.Microscopy.com} Sent: Wednesday, January 12, 2005 7:39 AM
Hey Jim!
Yes. I have owned directly or indirectly about 4 of these instruments over the past 30+ years. They were conceived in about 1969 by a company in California (ETEC Corp) and were the first serious challenge to the Cambridge Stereoscan and JEOL U2. They were considered the "Rolls Royce" of SEMs in their day until Jim Dow (I think that was his name) sold the company in the early 1980s. I believe Perkin-Elmer purchased it in order to make the first electron beam lithography devices The company (ETEC) still exists today as far as I am aware for this purpose. They no longer make SEMs.
It was innovative for its time in that it was of a completely modular design where whole functionalities could be exhanged overnight as nim-bin modules. As an SEM it only used W filaments.
Also, as far as I know, there are still a fair number of operating models out there. All mine I am afraid have been replaced and/or donated but one I gave to Puerto Rico was working (with a lot of TLC) until about 2 years ago. FYI, there is a person who has a company who deals in spares and rebuilds ETECs - his name is Gary Easton and his company is "Scanners Company" in Maryland somewhere (Google to find out!) Also another fellow is Hank Bebe who works for the Rich Lee Group who knows as much as anyone about the Autoscan.
I haven't checked Google to verify my memory of all the above but I don't believe I am far off!
Please feel free to give me a call if you would like to discuss more. There are some fun anecdotes concerning this instrument and the ETEC company!
Cheers etc
Peter
} - } } Hi all, } } I'm looking at a paper that did some work with and Etec Autoscan. I'm not } very familiar with the instrument, and no model number is given. Can anybody } fill me a bit on the history of this SEM? What is it's vintage, W, } Lab6 or FE, etc.? } Does the company exist under another name? All the images and info I can find } on the web look to be a 70s to 80s type instrument, but... } } Gee, a "History of Microscope Manufacturers: Intrigue, Bankruptcy } and Hostile Takeovers" } web site would be handy... } } As usual, thanks in advance! } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/dmf
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
Thanks to all who sent the many detailed descriptions of the Etec Autoscan. I have more than enough information for what I was curious about. But it's always good to read about scopes that people have known and loved. This looks to be true for the Autoscan. I think the subject has come up before, but it would be nice to have a repository of images and specifications for the various instruments through the ages. Oops! Sounds like I just volunteered. But if those interested would like to forward me info about their favorite (or most dreaded) scope and interesting anecdotes, horror stories, etc. I'll see what I can do about putting a "Rogue's Gallery" on my website. Any interest out there?
Thanks again,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
The website idea isn't bad, but it would be very frustrating to put together, much less maintain. I'd host it, though, if anyone wants to help.
ETEC is a company that grew in the 70's, started by some engineers from MAC (you can still see some MAC microprobes around). They were very successful and made some solid, well engineered SEMs. One of the more interesting aspects is the wide range of accessories they had available. These instruments were extreme research machines and anything they could think of making for it, they did. There was another model, the Omniscan, that had some cute ideas but ended up being a maintenance nightmare. For the most part, they used tungsten cathode, although LaB6 became available around the end (the first to offer it that I know, the tips didn't last long, though). They were also developing a variable pressure SEM that never made it to market.
The end, by the way, was around 1983. ETEC had been developing Electron Beam Lithography (EBL) for years. In 1979, they were bought by Perkin-Elmer, who wanted the EBL component, but didn't care about the laboratory instruments. They simply let the SEM manufacturing die a slow death, selling off existing inventory, without ever really letting customers know.
The ETEC EBL was a raster-based device. An elaborate table supported by air bearings held the wafer, which was stepped around to allow the beam to expose each small square area of the wafer. Since it had to store the pattern for an entire wafer at a resolution of less than 100nm, large amounts of memory were needed. Shortly after being bought by Perkin-Elmer, some Japanese manufacturer's came in with vector based EBL systems and pretty well trounced ETEC.
ETEC lives on today as an independent (I think) manufacturer of EBL.
I left ETEC in 1982. It was becoming obvious that they were winding down the SEM business and trying to pressure me into the EBL lines. They virtually sold you a service engineer along with the instrument - he'd move where the instrument went and be on 24/7 call. Now, some SEM customers can be rather demanding, but a semiconductor manufacturer loosing millions of dollars every hour an instrument is down can be a real pain. Didn't sound like too much fun to me.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Wednesday, January 12, 2005 9:39 AM, James M. Ehrman [SMTP:jehrman-at-mta.ca] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Hi all, } } I'm looking at a paper that did some work with and Etec Autoscan. I'm not } very familiar with the instrument, and no model number is given. Can anybody } fill me a bit on the history of this SEM? What is it's vintage, W, Lab6 } or FE, etc.? } Does the company exist under another name? All the images and info I can } find } on the web look to be a 70s to 80s type instrument, but... } } Gee, a "History of Microscope Manufacturers: Intrigue, Bankruptcy and } Hostile Takeovers" } web site would be handy... } } As usual, thanks in advance! } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/dmf }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 13 17:20:59 2005
This position has come to my attention, and I thought it might be of interest to the broader SEM community.
Regards,
Peter
Peter McSwiggen McSwiggen & Associates, P.A. 2855 Anthony Lane South, Suite B1 St. Anthony, MN 55418 phone: 612.781.2282 fax: 612.781.7540 e-mail: PMcS-at-McSwiggenAssoc.com
_________________________ Guidant is seeking a SEM/EDS technician to operate a Scanning Electron Microscope (SEM). This is a full time, 1st shift position in Guidant's Corporate laboratory. Minimum requirements include: A two year technical degree and 2 - 4 years of experience in Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). Ability to work independently and in a team oriented environment. Good writing skills with an ability to independently write reports using standard and custom software. Preferred qualifications include experience with surface and failure analysis of electronic components in a FDA regulated environment. Knowledge of material properties and characteristics is a plus.
For more information or to apply contact:
Bruce Peacock Sr. Scientist, Corporate Lab Guidant Corporation 4100 Hamline Ave N. St. Paul, MN 55112 Telephone: 651.582.2075 bruce.peacock-at-guidant.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 14 03:55:37 2005
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Hi all, I have a user who wants to stain lignin in JB-4 resin-embedded corn stalks. He tried Phloroglucinol which works fine in paraffin embedded samples since the paraffin is removed. However, this stain requires an HCl treatment to reveal the desired color. The acid treatment interacts adversely with the JB-4 resin.
The resin embedding gives much better resolution than the paraffin so it is not an option to go back to the paraffin-embedded tissue.
Does anyone have an alternative stain that will work with this resin?
Thanks in advance, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 14 04:05:19 2005
I thought that it was about mid 60's and Norma Reid's text on ultramicrotomy cofirms that it was 1965. It was an improvement on the 'Ultratome I' because it had finer manual cutting range and extended cutting stroke as well as upgraded optics and a few other things. There was never an 'Ultratome II' because this was the designation for the upgrade kit for the I which turned it into something like a III. We still occasionally use a I with a part II kit modification.
Ref Ultramicrotomy, Norma Reid (1975) (Part II of Vol 3 Practical Methods in Electron Microscopy - Ed Audrey M. Glauert).
Hope this answers your question without giving you too much information.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Ephram Shizgal {shizgal-at-lv-em.com}
Hi
Interesting about the Ultratome III coming out in '65. We bought one around 1978 I think and then after I had changed jobs, a IV in about 1984. So the III was around for a long time.
Gareth
At 11:05 2005-01-14, Malcolm Haswell wrote:
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Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 14 08:47:53 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lbalakrishnan-at-medicine.nodak.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, January 13, 2005 at 11:03:29 ---------------------------------------------------------------------------
Email: lbalakrishnan-at-medicine.nodak.edu Name: Lata Balakrishnan
Organization: University of North Dakota
Education: Graduate College
Location: Grand Forks, North Dakota, USA
Question: Hi, I have been trying to visualize SV40 chromatin using TEM. Our department does not have a glow discharge apparatus. So I am unable to do a glow discharge before applying my samples to the carbon coated formavar grid. Is there any other method that alternates the glow discharge so that the chromatin will adhere to the grid. At this point I am unable to pick up any chromatin just by rotary shadowing and tungsten coating. Also during tungsten coating sometimes the grid fries up and is compeletely broken. Any suggestions?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fahayes-at-comcast.net) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 13, 2005 at 12:50:22 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (derek.dunfield-at-gmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 13, 2005 at 19:28:08 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: Long Distance DIC?
Question: Hello all, Currently my lab is looking for a long distance (around 13mm or more depending on the shape of the objective - the longer the better) DIC or phase lens with high magnification (50X - 100X, again bigger is better). The idea is to use this lens with electrophysiology tools -hence the need for the long working distance. Do they exist? Any help would be most appreciated!
We are buying DVDs from a "namebrand" manufacturer for storing copies of our digital data. I know that this discussion has gone around before, but I am still curious.
I can buy "4X" speed DVD-Rs for about $1 each. The same namebrand offers "8X" speed DVD-Rs for half the price, which presumably would allow us to write or read the data twice as fast. And of course off-brand DVDs are sold for much less.
I want to write data safely and reliably, so I'm paying for the namebrand. Is it logical to also buy the slower access time DVD-Rs at the higher price? I have heard horror stories about losing data on cheaper media, or on media with the wrong style of writing implement used to mark the contents.
Is 8X a safe bet, or should I stay with slower media?
Have a good weekend. -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 14 16:44:29 2005
The act of having an image in focus in the TEM is when the objective lens places the image on the exact plane being viewed by the diffraction lens. Once these two lenses have been brought to this value the image will stay in focus no matter how many lenses we place between this point, known as the first image plane, and any viewing device. Possible solutions are outlined below
1) I suggest that you set focus on your viewing screen (which matches the two lenses as described above) and then adjust the other devices to bring them into focus. I am not sure how difficult this may be but the problem is not an instrument (microscope) focus problem.
2) The viewing device with the shorter specimen to device distance provides you with a lower magnification image than the device further away from the specimen. As this is the case the lower device would be more critical of the focus setting and maybe it is this that goes some way to that device appearing out of focus. If you focus the image on the screen at say 200,000X with a 10X binocular and then display it upon the TV camera at 100,000X does the more critical image focus solve the problem? This assumes that there is not a diffraction lens change between these two magnifications. Such a change would change the desired objective lens setting.
Good luck
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide www.emcourses.com tel +44 1280 816512 fax +44 1280 814007
----- Original Message ----- } From: "K.N. Bozhilov" {bozhilov-at-ucr.edu} To: {microscopy-at-microscopy.com} Sent: Wednesday, January 12, 2005 12:42 AM
David Hall writes ...
} I can buy "4X" speed DVD-Rs for about $1 each. The same namebrand } offers "8X" speed DVD-Rs for half the price, which presumably would } allow us to write or read the data twice as fast. And of course } off-brand DVDs are sold for much less. } } I want to write data safely and reliably, so I'm paying for the } namebrand. Is it logical to also buy the slower access time DVD-Rs } at the higher price? I have heard horror stories about losing data } on cheaper media, or on media with the wrong style of writing } implement used to mark the contents.
You are correct regarding "cheap" media, and you are somewhat safe buying name brands, but not entirely, because you can buy the same name brand one month and realize it is made in Japan, and buy the same again the next month and realize after it is made by a different process in Taiwan.
Many will also imply the safest write speed is the slowest, ... e.g., to purchase any reputable media and always write at 2.4x ... with patience, and enable verification after the write. It's your data!
Hi all, We have a position available shortly in our unit - see details below. The selection criteria etc should be available at the given web address next week, or email Alanah.McCann-at-anu.edu.au Briefly, we are a campus-wide unit doing approximately 60% materials work and 40% biological, 3 TEMs, 4 SEMs, a FIB/SEM, 3 EDXAs and light microscopes, cryo-gear. The work will involve user support, instrument maintenance and development.
Regards Sally Stowe
Dr SJ Stowe Facility Coordinator ANU Electron Microscopy Unit The Australian National University www.anu.edu.au/EMU/index.htm sally.stowe-at-anu.edu.au GPO Box 475,Canberra ACT 2601 Australia ANU CRICOS#00120C
Salary Package: $39,341 - $47,861 pa plus 17% Super
We are seeking a highly motivated person to join the staff team in a cross-disciplinary microscopy facility. Knowledge and expertise in fields relevant to scientific instrumentation such as electronic or mechanical engineering, computing and image analysis would be an advantage, as would direct experience with electron microscopy. The web address for the unit is www.anu.edu.au/EMU/index.htm Selection Criteria: http://info.anu.edu.au/hr/Jobs/General_Positions/_PDF/QESS .pdf or email: Alanah.McCann-at-anu.edu.au, phone 6125 4138.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ronpeters-at-integraonline.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, January 16, 2005 at 16:01:22 ---------------------------------------------------------------------------
Email: ronpeters-at-integraonline.com Name: Ron Peters
Education: Graduate College
Location: Prior Lake, MN, USA
Question: I am confused about the exact difference between objectives marked "Oil" immersion and those marked Homogenous Immersion ("HI"). What exactly are the differences in applying these two different objectives? Does an HI objective use a cover slip? Does an HI objective use the same immersion oil as an "Oil" objective?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ronpeters-at-integraonline.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, January 16, 2005 at 16:01:22 ---------------------------------------------------------------------------
Email: ronpeters-at-integraonline.com Name: Ron Peters
Education: Graduate College
Location: Prior Lake, MN, USA
Question: I am confused about the exact difference between objectives marked "Oil" immersion and those marked Homogenous Immersion ("HI"). What exactly are the differences in applying these two different objectives? Does an HI objective use a cover slip? Does an HI objective use the same immersion oil as an "Oil" objective?
I found from experience that the side port (35mm port) on most TEMs is not parfocal with the plate camera film or the bottom port. Some TEMs are close enough, others are not. There are a few exceptions, most notably the JEOL-100C (and 200C), which have side ports that are dead-on parfocal with the focusing screen, plate camera film, and bottom port.
The whole issue arises in the following cases:
1) A side mounted CCD camera (instead of binoculars and focusing screen) is used to focus for a film plate camera (a bad idea unless the camera is low view angle and side port is parfocal with the plate film).
2) A TEM focusing screen with binoculars is used to focus for a side mounted slow scan camera that has no live focus capability.
3) A bottom mounted TV camera is used as a focusing aid for a side mounted slow scan CCD. Such configuration is not common.
Of course, cases (2) and (3) are no problem for a 1K x 1K or fewer pixel count side mounted CCD camera with a wide viewing angle, but a side mounted camera of more than 2 megapixels had better have a live focus capability. Otherwise, some focus correction will probably be required compared to the focusing screen or bottom mounted TV camera. The higher the pixel density is, the more noticeable a focus difference becomes.
I know that the entire space below the final projector lens is considered to be parfocal (including several floors below the TEM :-). In practice, however, this is not always the case. I should confess that I didn't study this condition beyond accumulating TEM brand/model- specific statistics.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane Duluth, GA 30096 Tel. (770)232-7785 Fax (770)232-1791 Mobile (678)467-0012 www.sia-cam.com ----- Original Message ----- } From: "K.N. Bozhilov" {bozhilov-at-ucr.edu} To: {microscopy-at-microscopy.com} Sent: Tuesday, January 11, 2005 7:42 PM
Electron Microscopy Technician
The Materials Characterization Laboratory, part of Penn State's Materials Research Institute, has an immediate opening for an experienced electron microscopy technician. Responsibilities will include overseeing the maintenance on three transmission electron microscopes (TEM), one focused ion beam (FIB) and a variety of support equipment. Applicants should have hands-on experience operating and repairing electron columns, vacuum systems and electronics. Other responsibilities will include maintaining sample preparation equipment such as diamond saws, ion thinners, polishing equipment as well as training users in the operation of that equipment.
MINIMUM QUALIFICATIONS: Associate's degree in a technical or administrative program, or equivalent knowledge, plus 1 year related work experience.
Send resume and cover letter (by e-mail, USPS, or courier) to: Joe Kulik The Pennsylvania State University 194 Materials Research Institute Building University Park, PA 16802-7003
e-mail: juk12-at-psu.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 19 04:36:10 2005
Did anyone ever answer your question? You asked, "Why don't we see the monotonic rise as described by "self-biasing", but instead hills and valleys?"
I would be interested in reading the answers you received to this question.
Since we can't post attachments, I looked for some slides on the internet for me to discuss how this circuit works (between looking for a position in microscopy). I was pleasantly surprised to find a PDF file at my old alma mater, Ohio State University, in the geology department no less. See pages 17 and 18 below. http://www.geology.ohio-state.edu/~bhattiprolu/GS675/notes/lecture1.pdf
This PDF file shows a general characteristic curve shape on page 18.
My background in electronics, as related to microscopy and other areas, is below.
Paul Beauregard Chemist (Ohio State) & Microscopist. Electronics, University of Akron, Summa Cum Laude. FCC licensed commercial radiotelephone license holder. FCC extra class license holder, station call sign is KC8O /3. • — • — • • • • — • —
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 19 10:01:44 2005
I don't know how much stock I would place in the Ohio State set of notes since they name wavelength as the chief factor in SEM resolution instead of probe size.
Franklin Bailey University of Idaho Electron Microscopy Center
-----Original Message----- } From: Beaurega [mailto:beaurega-at-westol.com] Sent: Tuesday, January 18, 2005 11:11 AM To: michael shaffer; MSA listserver
Dear Michael,
Did anyone ever answer your question? You asked, "Why don't we see the monotonic rise as described by "self-biasing", but instead hills and valleys?"
I would be interested in reading the answers you received to this question.
Since we can't post attachments, I looked for some slides on the internet for me to discuss how this circuit works (between looking for a position in microscopy). I was pleasantly surprised to find a PDF file at my old alma mater, Ohio State University, in the geology department no less. See pages 17 and 18 below. http://www.geology.ohio-state.edu/~bhattiprolu/GS675/notes/lecture1.pdf
This PDF file shows a general characteristic curve shape on page 18.
My background in electronics, as related to microscopy and other areas, is below.
Paul Beauregard Chemist (Ohio State) & Microscopist. Electronics, University of Akron, Summa Cum Laude. FCC licensed commercial radiotelephone license holder. FCC extra class license holder, station call sign is KC8O /3. • — • — • • • • — • —
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 19 11:35:44 2005
} I don't know how much stock I would place in the Ohio State set of notes } since they name wavelength as the chief factor in SEM resolution } instead of probe size.
One of the primary factors which minimize the spot size is "diffraction aberration", a function of wavelength ... is it not(?)
} -----Original Message----- } From: Beaurega [mailto:beaurega-at-westol.com] } Sent: Tuesday, January 18, 2005 11:11 AM } To: michael shaffer; MSA listserver } Subject: [Microscopy] Re: RE: RE: RE: SEM: gun saturation } } } } } ------------------------------------------------------------------ } ---------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------ } ---------- } --- } } Dear Michael, } } Did anyone ever answer your question? } You asked, "Why don't we see the monotonic rise as described by } "self-biasing", but instead hills and valleys?" } } I would be interested in reading the answers you received to this } question. } } Since we can't post attachments, I looked for some slides on the internet } for me to discuss how this circuit works (between looking for a } position in } microscopy). I was pleasantly surprised to find a PDF file at my old alma } mater, Ohio State University, in the geology department no less. } See pages } 17 and 18 below. } http://www.geology.ohio-state.edu/~bhattiprolu/GS675/notes/lecture1.pdf } } This PDF file shows a general characteristic curve shape on page 18. } } My background in electronics, as related to microscopy and other areas, is } below. } } Paul Beauregard } Chemist (Ohio State) & Microscopist. } Electronics, University of Akron, Summa Cum Laude. } FCC licensed commercial radiotelephone license holder. } FCC extra class license holder, station call sign is KC8O /3. } • — • — • • • • — • — } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 19 19:20:43 2005
Dear Colleague I am working on article regarding holey film preparation technique I succesfully use for two decades. I want to present historical data on this issue and sort of summary on different techniques used in EM. Unfortunately, most of the work on holey film preparation done in 50-60es and is not indexed in modern databases and my personal archive was lost when I moved to US. So, I could not restore some important references. I would greatly appreciate your help in pointing on old references/articles on holey film preparation I could cited/used in my work. I would be happy to share the information with EM community. Thanks for your help in advance, Sergey
P.S. Any suggestions where I could publish such work (with detailed instruction how to make holey films) would be greatly appreciated also. Have a great day.
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 20, 2005 at 20:31:36 ---------------------------------------------------------------------------
I have a feew question regarding petrographic microscope. does anyone know the process of preparing petrographic sample for imaging ? Please kindly share with me your valubale knowledge.
We are hiring a facility manager for the Imaging and Microscopy Facility at UC Merced (primarily for electron microscopy). UC Merced is the newest campus in the University of California system, located approximately 2 hours SE of the San Francisco Bay Area, and will open to 1,000 students in the Fall of 2005. Eventual enrollment is expected to be 25,000 students. Please see the job posting below for more details.
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We need to Epon embed and section neurons growing attached to a membrane. In the past (relatively long ago), we used to use millipore membrane inserts treated with poly-Lys. The neurons stuck very well to the membrane and more over, the membrane "curled" nicely during the processing giving us effectively larger surface cut on each section and so better chance to find what we needed. Currently, our neurons refuse to stay on the membrane and in addition, the membrane stays flat. I remember a discussion about the topic some time ago, but have problems to find it in the archives. Does anybody have a good idea?
Thanks,
Michael Jarnik, FCCC, Philadelphia
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 21 11:31:07 2005
The simple answer is that, typically, a slab of rock is cut off with a diamond saw, glued to a petrographic slide, then ground and polished to an appropriate thickness (ex: 30microns). If it is a mineral that contains quartz, the 30micron thickness is evaluated by its color between crossed polars (146nm retardation). For more complete information and tips regarding specific minerals, I'd recommend your contacting Buehler, Struers, Logitech, or Mark V Labs. They all have the necessary equipment, supplies and expertise.
One other note: petrographic slides are not the same size as normal 1"x3" microscope slides.
Hope this was helpful,
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 09:33 PM 1/20/2005, kssim-at-mmu.edu.my wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Jan 21 20:16:25 2005
Hi Sergey, Do you have access to Desmond Kay's Techniques for Electron Microscopy, 2nd edition, 1965? If so, Chapter 3 was written by D. E. Bradley , pg 58-74, on The Preparation of Specimen Support Films and has early references for it. It covers several of the early techniques.
Kay, Desmomd (Ed), 1965, Techniques for Electron Microscopy, F. A. Davis, Co., Philadelphia, PA
If you do not have access to it, I can scan in the references for you and send them to you off line.
If it is a new technique for making holey grids, then perhaps the Microscopy and Microanalysis Journal which is the official journal of the Microscopy Society of America (see microscopy.com and follow information to the journal). Also, you might consider Journal of Microscopy Research and Techniques. If it is just a short article, you might consider Microscopy Today (information also available at microscopy.com).
Best of Luck, Judy Murphy Stockton, CA
Sergey Ryazantsev wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Dear Colleague } I am working on article regarding holey film preparation technique I } succesfully use for two decades. I want to present historical data on } this issue and sort of summary on different techniques used in EM. } Unfortunately, most of the work on holey film preparation done in } 50-60es and is not indexed in modern databases and my personal } archive was lost when I moved to US. So, I could not restore some } important references. I would greatly appreciate your help in pointing } on old references/articles on holey film preparation I could } cited/used in my work. I would be happy to share the information with } EM community. Thanks for your help in advance, Sergey } } P.S. Any suggestions where I could publish such work (with detailed } instruction how to make holey films) would be greatly appreciated } also. Have a great day. } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-080 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } } }
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From MicroscopyL-request-at-ns.microscopy.com Sat Jan 22 11:48:15 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (germaine_g_boucher-at-groton.pfizer.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 21, 2005 at 13:07:37 ---------------------------------------------------------------------------
Hi Germaine, Normally we dealt with sections of paraffin blocks as we wanted a specific area.
For fiber and mineral content: 1. We used a conventional 6 micron section from a paraffin block. 2. We mounted it on a glass slide treated it with xylene and alcohol to remove the wax.
For fiber and mineral content: If we were doing fiber analysis in the sample, we allowed it to dry, ashed it in a muffle furnace at 450 degrees C until the tissue was completely ashed. 3. Added polyvinyl alcohol (PVA) solution to the specimen area, and allowed it to dry. 4. Peeled off the hardened film. 5. Placed hardened film upside down on a slide. 6. Evaporated carbon on upper face of hardened film. 7. Floated carbonized specimen onto the surface of hot water. This dissolves the water-soluble plastic and leaves the carbon film and the contained ashed tissue floating on the surface. 8. Gently, broke up the film into 3 mm squared pieces. 9. Picked up pieces on EM grid and examined
This was used way back in the seventies by F.C.Pooley and details published by Langer, A.M. et al, 1972. Chemical Characterization of Uncoated Asbesots Fibers from the lungs of Asbestos Workers by Electron Microproble Analysis. J. Histochem. Cytochem 20:735.
To re-embed for TEM: After 1 and 2 above 3. Once in alcohol, changed 2 or 3 times to be sure it was dehydrated in 100% EtOH. 4. For Epon Resin Mixture: further dehydrated in propylene oxide, 2 times 10 min each on the slide. 5. Added graded series of PO:Epon replacement mix(2:1; 1:1; 1:2) for 30 min each 6. Changed to pure epon mix with 2 changes for 30 min each, on the slide 7. Took off excess resin from slide with sample, and put a full capsule (full of epon mix) over the slide. 8. Put in oven (60C) and hardened it. 9. After hardening, can pop it off by putting in liquid nitrogen quickly and one gets the section at the tip of the capsule. 10. We developed a slide holder (sold yrs ago by Polysciences) to hold the slide while it was easily popped off, but anything will do just so it is held so the pressure point is at the capsule/slide interface.
NOTES: A. If LRWhite is used, the propylene oxide was not used and we prevented air from getting to the mixture when being polymerized. Have occasionally used propylene oxide by mistake over the yrs and that seems to work as well with LR White but generally do not use it with LR White. With LR White, we hardened at 52C if we were not in a hurry or 1 hr at 90C, if we had to do the same day. B. Most EM Supply companies sell Epon substitute (Ted Pella, EMS, Fullam, SPI, etc.) We used Glauerts medium mix for hardness. All Epon mixes had the catalyst in them when used. C. In later yrs , we used the microwave for the later stages after the paraffin was gone. Paraffin is transparent to microwaves so doesn't melt in the microwave, just like a dry ice cube doesn't melt because it also is transparent to the microwaves! (As an aside, that is a great bar trick! - before you bet, just be sure the ice cube is dry of moisture which acts as a catalyst for melting). If you use a microwave, use one with a cooled specimen stage. The microwave speeds up the process tremendously and it is done in no time and one can section and do the analysis the same day easily with time to spare.
I have not tried the entire paraffin block as usually I had specific areas I was looking for so had to section the paraffin block to find out where I wanted to look. It would seem that the same process would work, however I would think the times would have to be increased somewhat. Of course, as you know, section or block, the ultrastructure is only as good as the fixation and preparation was initially when the sample was fresh!!!
Hope that is helpful, Judy
Judy Murphy, PhD Microscopy & Imaging Consultant Stockton, CA murphyjudy-at-comcast.net
by way of MicroscopyListserver wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Jan 22 19:30:06 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pilttdownman-at-wmconnect.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, January 22, 2005 at 18:06:32 ---------------------------------------------------------------------------
Email: pilttdownman-at-wmconnect.com Name: Ron Joyner
Organization: Mississippi Gulfcoast community college
Education: Undergraduate College
Location: Gautier, Mississippi
Question: what advantage does a cardioid condensor have over a star diaphragm?
Hi, This is a post script for the procedure below for deparaffinization of materials.
If one is doing morphology the below procedure which is listed below will work however for quantitative numbers of fibers which must be identified particularly by diffraction, plasma ashing should be used to prevent the asbestos from changing its crystallinity structure which of course prevents a correct diffraction pattern.
It is good to have good friends that help point out missing information especially important missing information. Thanks Chuck,
Judy Murphy
Judy Murphy wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Reprocessing paraffin blocks } } Hi Germaine, } Normally we dealt with sections of paraffin blocks as we wanted a } specific area. } } For fiber and mineral content: } 1. We used a conventional 6 micron section from a paraffin block. } 2. We mounted it on a glass slide treated it with xylene and alcohol } to remove the wax. } } For fiber and mineral content: } If we were doing fiber analysis in the sample, we allowed it to dry, } ashed it in a muffle furnace at 450 degrees C until the tissue was } completely ashed. } 3. Added polyvinyl alcohol (PVA) solution to the specimen area, and } allowed it to dry. } 4. Peeled off the hardened film. } 5. Placed hardened film upside down on a slide. } 6. Evaporated carbon on upper face of hardened film. } 7. Floated carbonized specimen onto the surface of hot water. This } dissolves the water-soluble plastic and leaves the carbon film and the } contained ashed tissue floating on the surface. } 8. Gently, broke up the film into 3 mm squared pieces. } 9. Picked up pieces on EM grid and examined } } This was used way back in the seventies by F.C.Pooley and details } published by } Langer, A.M. et al, 1972. Chemical Characterization of Uncoated } Asbesots Fibers from the lungs of Asbestos Workers by Electron } Microproble Analysis. J. Histochem. Cytochem 20:735. } } To re-embed for TEM: } After 1 and 2 above } 3. Once in alcohol, changed 2 or 3 times to be sure it was dehydrated } in 100% EtOH. } 4. For Epon Resin Mixture: further dehydrated in propylene oxide, 2 } times 10 min each on the slide. } 5. Added graded series of PO:Epon replacement mix(2:1; 1:1; 1:2) for } 30 min each } 6. Changed to pure epon mix with 2 changes for 30 min each, on the slide } 7. Took off excess resin from slide with sample, and put a full } capsule (full of epon mix) over the slide. } 8. Put in oven (60C) and hardened it. } 9. After hardening, can pop it off by putting in liquid nitrogen } quickly and one gets the section at the tip of the capsule. } 10. We developed a slide holder (sold yrs ago by Polysciences) to } hold the slide while it was easily popped off, but anything will do } just so it is held so the pressure point is at the capsule/slide } interface. } } NOTES: } A. If LRWhite is used, the propylene oxide was not used and we } prevented air from getting to the mixture when being polymerized. } Have occasionally used propylene oxide by mistake over the yrs and } that seems to work as well with LR White but generally do not use it } with LR White. With LR White, we hardened at 52C if we were not in a } hurry or 1 hr at 90C, if we had to do the same day. } B. Most EM Supply companies sell Epon substitute (Ted Pella, EMS, } Fullam, SPI, etc.) We used Glauerts medium mix for hardness. All } Epon mixes had the catalyst in them when used. } C. In later yrs , we used the microwave for the later stages after } the paraffin was gone. Paraffin is transparent to microwaves so } doesn't melt in the microwave, just like a dry ice cube doesn't melt } because it also is transparent to the microwaves! (As an aside, that } is a great bar trick! - before you bet, just be sure the ice cube is } dry of moisture which acts as a catalyst for melting). If you use a } microwave, use one with a cooled specimen stage. The microwave speeds } up the process tremendously and it is done in no time and one can } section and do the analysis the same day easily with time to spare. } } I have not tried the entire paraffin block as usually I had specific } areas I was looking for so had to section the paraffin block to find } out where I wanted to look. It would seem that the same process would } work, however I would think the times would have to be increased } somewhat. Of course, as you know, section or block, the } ultrastructure is only as good as the fixation and preparation was } initially when the sample was fresh!!! } } Hope that is helpful, } Judy } } Judy Murphy, PhD } Microscopy & Imaging Consultant } Stockton, CA } murphyjudy-at-comcast.net } } } } by way of MicroscopyListserver wrote: } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (germaine_g_boucher-at-groton.pfizer.com) from } } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, } } January 21, 2005 at 13:07:37 } } --------------------------------------------------------------------------- } } } } } } Email: germaine_g_boucher-at-groton.pfizer.com Name: Germaine Boucher } } } } Organization: Pfizer } } } } Title-Subject: [Microscopy] [Filtered] MListserver: } } } } Question: Does anyone out there have a protocol for reprocessing } } specimens from paraffin blocks for TEM? } } } } --------------------------------------------------------------------------- } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 23 08:45:24 2005
Thanks for clarify my about the petrographic microscope. It is valuable information.
Again thanks for helping me Ks ----- Original Message ----- } From: "Barbara Foster" {bfoster-at-mme1.com} To: "by way of MicroscopyListserver" {kssim-at-mmu.edu.my} ; {microscopy-at-microscopy.com} Sent: Sunday, January 23, 2005 1:32 AM
Colleagues, can anybody provide the formula/composition for Ilford PERCEPTOL B/W developer ? Obviously the photochemical department of ILFORD (UK) has filed {se?lp=ende&p=/Mn4k.&search=file} a {se?lp=ende&p=/Mn4k.&search=a} petition {se?lp=ende&p=/Mn4k.&search=petition} in {se?lp=ende&p=/Mn4k.&search=in} bankruptcy and I can't get / order the above cited developer anymore here in Germany. {se?lp=ende&p=/Mn4k.&search=bankruptcy} I need this developer for developing ("tenderize") the B/W ortho-films I use with my ZEISS EM109 TEM. Thanks for any advice! Peter Heimann
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 24 08:29:34 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ecd10-at-psu.edu) from http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html on Monday, January 24, 2005 at 08:15:03 ---------------------------------------------------------------------------
Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Penn State
Title-Subject: [Microscopy] [Filtered] MListserver: Post-doc Opening at Penn State
Question: POST-DOCTORAL POSITION in Analytical Transmission Electron Microscopy at The Pennsylvania State University
A postdoctoral position is available in the area of analytical transmission electron microscopy beginning March 1, 2005. The research project focuses on understanding structure and chemistry of amorphous metal oxides for capacitor and IR sensor applications. Through a variety of electron imaging, spectroscopy and diffraction (e.g. fluctuation EM) techniques we aim to quantify structure and chemistry of amorphous metal-oxide layers to help establish processing/structure/property relationships for this class of materials. Furthermore, we aim to understand the microstructural and microchemical evolution under applied bias. Most of the research will be conducted on a JEOL 2010F Field Emission TEM/STEM outfitted with an EDAX energy dispersive x-ray spectrometer (EDS), Gatan Enfina electron energy loss spectrometer (EELS), high-angle annular dark field STEM detector, and acquisition hardware and software for spectrum imaging. The ideal candidate for this position will have experience in HREM, STEM, EDS and EELS. The salary will be commensurate with qualifications and experience. Exceptionally qualified candidates may be considered at the Research Associate level.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
There was a well-known British photographer who died suddenly last year at a relatively young age, worked mostly (exclusively?) in B&W, can't remember his name, Barry something or other? I have his book at home, I think he used his own version of Perceptol, I will check at home tonight. I did not find Perceptol in Steve Anchell's books.
Geoff
PETER HEIMANN wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Colleagues, } can anybody provide the } formula/composition for Ilford PERCEPTOL B/W developer ? } Obviously the photochemical department of ILFORD (UK) has filed } {se?lp=ende&p=/Mn4k.&search=file} a {se?lp=ende&p=/Mn4k.&search=a} } petition {se?lp=ende&p=/Mn4k.&search=petition} in } {se?lp=ende&p=/Mn4k.&search=in} bankruptcy and I can't get / order the } above cited developer anymore here in Germany. } {se?lp=ende&p=/Mn4k.&search=bankruptcy} } I need this developer for developing ("tenderize") the B/W ortho-films } I use with my ZEISS EM109 TEM. } Thanks for any advice! } Peter Heimann } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 24 09:44:11 2005
My name is Erin Wagner. I am currently a graduate student at the University of the Pacific in Stockton, Ca. Recently, I have been reviewing the literature for my graduate project and have found some difficulty in retrieving the information I am looking for. While researching on-line I ran across your website. I was wondering if you would be willing to help me find the information I have been looking for.
For my project I am studying a protein that may be involved in cross-linkage and we would like to confirm this using SEM. However, the protein is embeded in a fiber and the only way to expose the protein is to solubilize the fiber in guanine-HCL. The fiber can be transferred to a solution less harsh such as 50 mM Tris-HCL by dialysis but the fiber is not soluble in water. I was wondering if SEM is possible under such conditions and if so whether you might know a protocol or where I could find a protocol to conduct the experiment. All the papers I have found discuss growing tissue on a slide, since my sample is a protein on a fiber that can only be exposed in guanine-HCL I can not grow it on a slide. Also, I think I would need to bind the primary antibody (which is a polyclonal antibody) in the tris-Hcl then bind a secondary antibody with the gold, maybe after fixation. I am unsure which fixation technique would be best and what size of gold to use.
Any help you could give me would be greatly appreciated. Thank you, Erin Wagner {mailto:e_wagner-at-pacific.edu} e_wagner-at-pacific.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 24 11:01:40 2005
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! 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.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 24 18:11:23 2005
Erin It would be much easier to answer your questions if will be more specific: what is the protein, which fibril - how your protein "embedded" into fibril? Does fibril formed from the protein?
Basically, I think the best technique in your case would be TEM with negative staining. Antibodies (AB) will not interact with specific antigen in guanidine-HCl - this chemical is strong denaturing agent, it'll equally denature (read deactivate) most of the ABs. If your protein somehow present on fibril or part of the fibril- then ABs will react and you may visualize the complex by negative staining. You may also enhance the signal using gold-conjugated secondary ABs (10 nm would be OK). Basically, SEM may be useless based on information you provided.
Have a good day, Sergey
At 07:49 AM 1/24/2005, 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 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Many years ago, this was treated as Kodak Microdol-X. I used either one interchangeably. They were pretty much the same to me. Temperature and freshness were critical.
I figure that all b/w developers (other than Microdol, et. al.) are variances of the basic chemistry of b/w developers. Check out the following links...
Ilford b/w film seemed to turn out better with Microdol for either FP4+ or HP4+. But the difference for results was the Zone treatment. In this case, over exposure and underdevelopment resulted in astonishing results. There is a huge amount of discussion about this that lingers to this day. The stated ISO of these films were not what I used.
If you want to see what Ilford FP4+ and HP5 do, relative to fine art, navigate your way through
http://www.photoweb.net
You will find a large set of photo images that are b/w and all shot using Ilford.
Why not underexpose and over develop? Well, if the info is not on/in the neg, over developing is not going to bring it out. This brings out the S curve about which Ansel Adams made history. For SEM, this is pre-collection LUT/gamma. No info at collection cannot be made up later on. And blown out highlights cannot be reduced to fact later on.
I guess that film is not dead yet. But the principles slowly migrate to digital.
gary g.
At 09:07 AM 1/24/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jan 25 02:56:58 2005
I am passing along a question regarding SEM sample preparation of a hard-soft specimen that I am hoping to get advice on. A colleague has implanted a collagen/titanium sample into bone and now removed it, and wishes to observe the interface of the bone with collagen, collagen with titanium, titanium with skin in the SEM. Any suggestions on sample preparation methods would be greatly appreciated. Further details of the sample are below.
It is 5 cm wide by 2 cm deep and the length is as follows:
/bone/collagen/titanium/skin {2 cm} / {5cm} / {4cm} (titanium and skin)
Thank you, Valerie -------------------------------------------------------- Valerie J. Leppert, Assistant Professor University of California, Merced School of Engineering
Mailing Address: P.O. Box 2039 Merced, CA 95344
Physical Address (for couriers/parcels): 4225 N. Hospital Road, Bldg 1200 Atwater, CA 95301
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Many years ago, this was treated as Kodak Microdol-X. I used either one interchangeably. They were pretty much the same to me. Temperature and freshness were critical.
I figure that all b/w developers (other than Microdol, et. al.) are variances of the basic chemistry of b/w developers. Check out the following links...
Ilford b/w film seemed to turn out better with Microdol for either FP4+ or HP4+. But the difference for results was the Zone treatment. In this case, over exposure and underdevelopment resulted in astonishing results. There is a huge amount of discussion about this that lingers to this day. The stated ISO of these films were not what I used.
If you want to see what Ilford FP4+ and HP5 do, relative to fine art, navigate your way through
http://www.photoweb.net
You will find a large set of photo images that are b/w and all shot using Ilford.
Why not underexpose and over develop? Well, if the info is not on/in the neg, over developing is not going to bring it out. This brings out the S curve about which Ansel Adams made history. For SEM, this is pre-collection LUT/gamma. No info at collection cannot be made up later on. And blown out highlights cannot be reduced to fact later on.
I guess that film is not dead yet. But the principles slowly migrate to digital.
gary g.
At 09:07 AM 1/24/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 25 03:03:17 2005
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Erin It would be much easier to answer your questions if will be more specific: what is the protein, which fibril - how your protein "embedded" into fibril? Does fibril formed from the protein?
Basically, I think the best technique in your case would be TEM with negative staining. Antibodies (AB) will not interact with specific antigen in guanidine-HCl - this chemical is strong denaturing agent, it'll equally denature (read deactivate) most of the ABs. If your protein somehow present on fibril or part of the fibril- then ABs will react and you may visualize the complex by negative staining. You may also enhance the signal using gold-conjugated secondary ABs (10 nm would be OK). Basically, SEM may be useless based on information you provided.
Have a good day, Sergey
At 07:49 AM 1/24/2005, 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 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (germaine_g_boucher-at-groton.pfizer.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, January 25, 2005 at 06:38:57 ---------------------------------------------------------------------------
Question: Thank you to everyone who responded to my question regarding reprocessing paraffin blocks for TEM. Your comments and suggestions have been helpful and enlightening.
Germaine Boucher TEM lab Pfizer Global Research and Development (860)715-2708
I am not sure I fully understand your experimental design but I differ with some of the comments that Sergey made. If you solublilize the protein in guanine-HCl, then allow it to dry down on a substrate, you could gently was with buffer, apply your primary antibody and follow that with a 5-10 nm colloidal gold conjugated secondary antibody. Smaller gold colloids react with higher efficiency than larger one (} 10 nm) so it is better to use a smaller label and then enhance the gold size so that it was easier to see in the SEM. I prefer gold enhanced gold (e.g., Nanoprobes kit) but silver enhanced gold is also ok. We do this for tissues in the SEM all the time with great results. Look at the gold with backscattered electron (BSE) detectors and the surface topography with secondary electron detectors. I am not sure why you need a fixation for an isolated fiber unless it was to bind it to the substrate; this would increase retention but lower immunoreactivity. Guanine-HCl treatment may destroy the reactivity of the epitope with the antibody but, on the other hand, it may increase reactivity similar to many antigen retrieval protocols.
At 09:49 AM 01/24/05, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
First, how was the sample stabilized i.e. in what solution, when it was removed and how is it being stored? Also, is there still a lot of soft tissue remaining?
Next do you have access to an environmental SEM or critical point dryer?
If the tissue is already dried, then rehydrating it in a fixative and standard biological SEM prep (given below), might help however there are certain interfaces that were likely damaged if air drying already occurred especially in the skin, as the pressure would be significant from the drying.
SO, if the sample were already dried, I likely would try the following:
1. Examine the sample carefully in the stereo microscope and get digital images and look for any possible drying artifacts (i.e. collapse between the epidermal cells and the Ti. Check carefully also the Ti and collagen. The collagen is fairly robust likely but the interface might have pulled away.
2. If you have an environmental, or low pressure SEM, then examine it directly in the SEM. This would be the easiest thing to do if the instrument is available.
If no ESEM or low pressure SEM is available
A. Fix the sample in 3% glutaraldehyde (buffered with cacodylate at pH 7.2), 1 hr. Agitate in a rotator during fixation. If the skin is thick, put in a vacuum chamber at room temp (i.e. a vacuum desiccator or vacuum oven). Usually 20 mm Hg pressure is enough. Pull the vacuum until all bubbles are gone from the vial, then turn off vac and let the sample fix. NOTE: Cacodylate (sodium salt) is usually a better buffer choice than phosphate for SEM as phosphates often precipitate.
B. Wash sample in buffer, 2 or 3 times
C. Fix in 2% aq Osmium tetroxide (in hood) for 1 hr, RT
D. Dehydrate in 50, 75, 95% EtOH, 2-10 min changes and 2-30 min changes in 100% EtOH
NOTE: If you have an industrial microwave, these times can be considerably shortened.
E. Critical point dry (CPD) sample using carbon dioxide.
F. Coat with conductive material e.g. AuPd while rotating the sample if possible.
If no CPD, then you could try putting it in xylene after the alcohol and air dry or dry it directly after the alcohol. The surface tension will be reduced somewhat when dried from an organic solvent, but of course there will be some shrinkage. In reality even CPD gives shrinkage, but usually less than drying from a solvent.
There is a method that uses HMDS (a silane), but don't think it is any more commercially available. People used the drying from a solvent and HMDS when they didn't have a CPD.
If the sample is already dried and you don't have a ESEM, then you may want to simply coat it and look at it. It somewhat depends on the number of samples you have. Once you coat it with a conductive coat, it will become more difficult to fix, however it still could be fixed from the bottom side if it isn't damaged from how it was mounted on the stub for viewing.
If you have a current field emission gun SEM, then you may also be able to use a low KV and get away with looking at it without coating. Many of the new SEMs have a low pressure mode, which could be used to view the sample also without coating.
The choice will be dependent on exactly what state the sample is in now, i.e. wet, or already dried.
Let me know if you need more details.
Good Luck, Judy
Judy Murphy, PhD Microscopy & Imaging Consultant Stockton, CA murphyjudy-at-comcast.net
vleppert-at-ucmerced.edu wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jan 25 18:07:35 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (psneeley-at-xmission.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, January 25, 2005 at 08:19:18 ---------------------------------------------------------------------------
Email: psneeley-at-xmission.com Name: P.S. Neeley
Organization: None
Title-Subject: [Microscopy] [Filtered] Looking for an AO Series 2 & 4 Microscope Reference Manual
Question: use an American Optical Series 2 scope (160mm tube length) and have been unable to find a copy of the reference (user) manual. These scopes (Series 2 and 4) were manufactured in the 1950s after the heyday of the ëblackí AO models, and before the advent of the infinity corrected AO models (Series 10, 20, 110, etc.). It is possible that you may still have some of these scopes, and their manuals, kicking around the lab or classroom someplace.
I would like to obtain the reference manual if possible.
Ultimately, I would scan the reference manual and place it with the other AO manuals and catalogs available at: http://www.xmission.com/~psneeley/Personal/Microscope.htm for anyone to access and use.
Thank you very much for the response. After careful consideration, my colleague, Dr. Hossein Hosseinkhani at ICYS, NIMS, Japan, tried the method suggested by Judy Murphy. He reports that it appears to have worked well and wanted me to convey his thanks for the suggestions.
Cheers, Valerie
-------------------------------------------------------- Valerie J. Leppert, Assistant Professor University of California, Merced School of Engineering
Mailing Address: P.O. Box 2039 Merced, CA 95344
Physical Address (for couriers/parcels): 4225 N. Hospital Road, Bldg 1200 Atwater, CA 95301
----- Original Message ----- } From: Judy Murphy {murphyjudy-at-comcast.net}
Microscopists,
Could anyone tell me how to measure and calculate the brightness and dose on the fluorescent screen of a transmission electron microscope? We have the Tecnai TEM.
Many thanks,
Lu
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 12:02:42 2005
Below is a question from a colleague (who is not on the list) regarding staining of organic carbon.
Dave Joswiak University of Washington
Hi,
I would be interested in any information regarding staining of organic carbon prior to observing the sample in a TEM. I work with meteorites and I am trying to distinguish the organic carbonaceous phases from the not-organic carbon present in the samples. I wonder if there exists any staining method to distinguish the organic carbon from the rest(it does not need to be specific to a type of organic carbon such as proteins, etc.). And I also wonder if there is a way/procedure of staining directly the TEM grids containing microtomed sections on them.
Thanks in advance.
-- Graciela Matrajt Dept. of Astronomy
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 14:03:15 2005
Hi, I do not know of a stain that would differentiate organic carbon from not-organic carbon. But am responding to the comment
"And I also wonder if there is a way/procedure of staining directly the TEM grids containing microtomed sections on them."
Most of the standard TEM section stains attach to proteins, nucleic acids, or unsaturated lipids or to the Os which reduced those unsaturated bonds. There are several specialty stains as well which can attach to for instance, certain sugars.
However the standard staining method for TEM sections is usually the Reynolds uranyl acetate, lead citrate stain or some derivative of that. Lead citrate can be made from components (lead nitrate, sodium citrate) or using the compound lead citrate directly (e.g. Venables instant lead citrate stain). Generally I have found that making lead citrate from its components is more stable for a longer amount of time than using the "instant" lead citrate stains, however I have used both successfully. When I use the "instant" lead citrate, I usually do so by making it up fresh each time.
Reynolds, E.S., 1963. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy, J. Cell Biol. 17, 208. Venable, J. H. and Coggeshall, R., 1965. A simplified lead citrate stain for use in electron microscopy, J. Cell Biol 25, 407. General reference Lewis, P.R., and Knight, D.P., 1977. Staining methods for sectioned material, in "Practical Methods in Electron Microscopy", Volume 5, Part I,Audrey Glauert (Ed), Elsevier/North-Holland Press.
Below is one way to do the Reynolds uranyl acetate, lead citrate stain. It is likely that the formatting will not be retained in the text mode so in addition I am sending you off line a word document as an attachment which will retain the format. The below method indicates keeping the stains in syringes in a charged desiccator however you can also keep the stains in the refrigerator, just so they are sealed in something that keeps especially the lead citrate away from air and UA away from light.
Especially if you are interested in trying to find something that will differentiate a very minute difference, it is suggested to use deionized, glass distilled water. I have found when working with a variety os polymer staining methods that water that is purified by a resin column method (e.g. millipore) tends to bring along some of the resin on occasion which also nicely stains.
If you find such a stain that can differentiate what you are looking for, would appreciate knowing about it.
Uranyl Acetate and Reynolds Lead Citrate Staining of Thin Sections for TEM: Preparation and Use Judy Murphy 050124
Staining of TEM thin sections involves a two step process, first staining with uranyl acetate and then with lead citrate. During the lead citrate staining, it is imperative to minimize the exposure of lead citrate with air to prevent precipitation on the sections of lead carbonate. To this end, NaOH pellets are used which removes the CO2 from the air. The more care that is taken to do the staining, the cleaner the sections will look.
NOTES: • Preparation of Reynolds Lead Citrate stain takes approximately 30-40 minutes. • Uranyl Acetate stain should be stored in a brown bottle to prevent reaction with light. • Both stock stains are stored in the refrigerator at 4OC. • Stains MUST be Millipore filtered just before use. • Small amounts of the stains may be stored in 3cc syringes with the filters attached and placed in a charged desiccator.
1. Boil deionized, distilled water as follow: NOTE: Freshly boiled water must be used to make the 0.02 N NaOH (if none available), water wash in wash bottle after staining, and to fill 10 ml water beakers at least 2 times. If new Lead Citrate stain needs to be made then it also needs to be made with freshly boiled water. A. Prepare Beaker as follows: a. Procure a clean 400-500 ml glass beaker. CAUTION: Use a beaker 2-3 times larger than the volume of water to be boiled b. Rinse the clean beaker with a small amount of deionized, distilled water. B. Fill the rinsed beaker half full with deionized, distilled water. C. Place beaker in middle of microwave and close the door. E. Microwave water for 10 mins. Exact time depends on the microwave. NOTE: Water should be allowed to boil about 5 minutes after it comes to a boil, which is about 10 min. in the microwave oven. CAUTION: Do not leave boiling water unattended as water may boil over. F. Using hot pad to protect skin, remove the beaker from the microwave. G. Cover the beaker with half of a petri dish. H. Allow the water to cool to room temperature. NOTE: It is important that as little CO2 which is found in the air be allowed to enter back into the water. For that reason, water should be boiled shortly before use, leaving enough time only to cool the water.
2. Assemble stains and staining materials while waiting for water: A. Freshly boiled and cooled deionized, distilled water (See Step 1 for preparation). B. 2% Uranyl Acetate (aqueous) stain in labeled, clean syringe with FRESH Millipore filter attached (See Step 5A for preparation). This is stored in a charged desiccator specifically for the stains. C. Reynold's Lead Citrate stain in labeled, clean syringe with FRESH Millipore filter attached (See Step 5B for preparation). This is stored in a charged desiccator specifically for the stains. D. NaOH pellets in sealed container. E. Wash bottle containing 0.02M NaOH (for rinse after Lead Citrate staining) a. Measure out 180ml of the freshly boiled and cooled deionized, distilled water. b. Pour water into a clean and rinsed wash bottle. c. Add 2 NaOH pellets. NOTE: Each pellet weighs approximately 0.07g d. Mix solution well but do not cause CO2 to enter. F. Wash bottle containing freshly cooled boiled distilled water. G. Assemble the following necessary materials on a lined tray: a. Three large glass Petri dishes (tops and bottoms). Line one Petri dish with a piece of filter paper indicated below. b. One small Petri dish. c. Three 10ml beakers. d. Two waste beakers, one for the 0.02 N NaOH rinse waste and one for the water rinse waste. e. Two squares of Parafilm cut to fit into one of the large Petri dish. f. 2 pieces of filter paper, one for lining the Petri dish. g. Filter paper arrows (fairly small arrows). h. Clean EM tweezers, preferably locking tweezers, if available (may use small o-ring to lock regular tweezers). i. Grid box containing sections to be stained with proper identification sheets.
3. Uranyl Acetate Staining: A. Put grids that are to be stained within 20 min. in a covered Petri dish lined with a piece of filter paper. B. Heat grids by putting them in the 52°C over for 15 minutes. A. Fill the 10ml beakers 3/4 full with boiled deionized, distilled water. B. Prepare Staining Petri dish as follows: a. Place a square of parafilm in the bottom half of a glass petri dish. CAUTION: Be sure that parafilm is flat. b. Place drops of 2% UA on the paraflim layer in the petri dish, one per grid to be stained. NOTE: Drops should be just larger than the grids. C. Float each of the grids, singly, section-side down on a separate drop of stain and cover the Petri Dish. D. Allow grids to stain for approximately 25-30 minutes. E. Remove the grids one at a time and immediately rinse by quickly dipping 20 times into each of the three10ml beakers of fresh deionized, distilled water. NOTE: Use a straight up and down motion. F. Holding the grid in the tweezers, remove the water retained on the grid by touching filter paper arrows to the edge of the grid. G. Place the grid on filter paper in a Petri dish. H. Clean up of UA: a. Dispose of stain in appropriate waste bottle under the fume hood. b. Dispose of solid waste (filter paper arrows, parafilm etc.) in appropriate bio-hazardous waste container.
4. Lead Citrate Staining: A. Fill the 10ml beakers approximately to 3 mm below the top, with freshly boiled and cooled deionized, distilled water. B. Prepare Petri dishes as follows: NOTE: Use a double Petri dish system i.e. small Petri dish top inside top and bottom of larger Petri dish. a. Place a square of Parafilm in the bottom half of a glass Petri dish. CAUTION: Be sure that parafilm is flat. b. Place 4 NaOH pellets close to but NOT touching where the Lead Citrate droplets will be placed. c. Place drops of Lead Citrate stain on the Parafilm layer in an area that can be covered by the small Petri dish, one per grid to be stained, near where the NaOH pellets are but NOT touching the pellets. NOTE: Drops should be just larger than the grids. d. Immediately put the small Petri dish top over the Parafilm area which has the Lead Citrate droplets and the NaOH pellets. Cover also the large Petri dish. e. Allow it to sit about 5 min. to remove CO2 from the atmosphere surrounding the stain. C. When ready to stain, quickly open the Petri dishes and float each of the grids, singly, section-side down on a separate drop of stain and quickly cover the Petri Dish. NOTE: Open the Petri dishes only as far as necessary to minimize the amount of air (which contains CO2) that gets into the Petri dishes. D. Allow grids to stain for appropriate time. This can vary between 5 to 20 min. NOTE: Stain time will vary. Perform test for type of tissue and stain. Staining Time Test: Start with 3 grids that all contain sections of the same thickness. These should be silver when viewed in the ultramicrotome. Stain one grid for 5 min, one for 10 min, and one for 15 min. with Lead Citrate. Uranyl acetate stain can all be 30 min. each. E. Rinse the grids one at a time as follows: a. Pick up the first grid and holding the grid with clean tweezers, gently run a stream of 0.02M NaOH (in wash bottle) over the grid and into a waste beaker. NOTE: Start the stream of NaOH first and then bring the grid into the stream so it doesn't put extensive washing force on the grid. b. Quickly, rinse the grid in the same manner with freshly boiled deionized, distilled water in a wash bottle. c. Immediately rinse the grid by quickly dipping at least 20 times into each of the three 10ml beakers of fresh deionized, distilled distilled water. NOTE: Use a straight up and down motion. F. Remove liquid on tweezers and grid as follows:] a. Holding the grid in the tweezers, remove the water retained in the tweezers by gently putting a paper arrow in between the tweezers coming from the non-pointed end downward. NOTE: Anti-capillary tweezers do not require this step. b. Still holding the grid in the tweezers, remove the water retained on the grid by carefully touching filter paper arrows to the edge of the grid. G. Place the grid on filter paper in a covered Petri dish to dry. After a few minutes, the grid can be placed in a grid box indicating proper labeling information on the Grid Box Information Sheet. H. Be sure to clean tweezers with clean water before picking up the next grid to be stained or before putting the tweezers away. I. Clean up Lead Citrate waste as follows: a. Dispose of used stain in appropriate waste bottle under the fume hood. b. Dispose of solid waste (filter paper arrows, Parafilm etc.) in appropriate biohazardous waste container.
5. Preparation of Stains: CAUTION: Wear Gloves when preparing stains. A. 2% aqueous Uranyl Acetate Stain: a. Measure 48ml deionized, distilled water into a clean100ml beaker. b. Weigh out 1 gram Uranyl Acetate. c. Add measured Uranyl Acetate to the deionized, distilled water in the beaker. d. Bring volume up to 50ml with deionized, distilled water. e. Stir solution on the magnetic stirrer to dissolve crystals. f. Store stock stain in properly labeled brown bottle. g. Prepare 3cc syringe of UA as follows: i. Using a 3cc syringe, draw about 2ml of UA. ii. Remove air from the syringe by holding the syringe straight up (needle on top) and slowly push on the plunger until the air is removed. iii. Place a clean Sweeney filter holder loaded with a NEW Millipore filter on the end of the syringe. iv. Label the syringe: "2% UA, Date and initials ". v. Wrap the syringe with tin foil to protect the stain from light. B. Reynold's Lead Citrate CAUTION: Lead stains precipitate as white opaque carbonate grains on grids if any contact with C02 is made. Be careful not to breathe on stain drops or on grids while rinsing. NOTE: Concentrated NaOH solutions should NOT be stored in glass-stoppered bottles or the stoppers will "freeze" and not be able to be removed. a. Pour 30ml of the boiled and cooled deionized, distilled water into a 50ml volumetric flask. b. Weigh out the following: i. Lead Nitrate crystals: 1.33 g ii. Sodium Citrate crystals: 1.76 g c. Add the Lead Nitrate and Sodium Citrate crystals to the 30ml of water in the volumetric flask. d. Mix the solution as follows: i. Tightly seal the top of the volumetric flask with parafilm. ii. Shake vigorously for 1 minute. NOTE: Solution should be milky white. iii. Allow to stand with occasional mixing for 30 minutes. e. Add 1 N NaOH until cloudiness disappears as follows: i. Add one drop of 1 N NaOH. ii. Swirl solution until mixed. iii. Repeat until cloudiness disappears. g. pH solution to pH 12 using 10 N NaOH and bring the volume up to 50 ml with freshly boiled and cooled deionized, distilled water. CAUTION: pH must be maintained above PH 12 or carbonate precipitate will form. h. Store stain is stored in a tightly stoppered bottle. i. Prepare 3cc syringe of Lead Citrate as follows: i. Using a 3cc syringe, draw about 2ml of Lead Citrate. ii. Remove air from the syringe by holding the syringe straight up (needle on top) and slowly push on the plunger until the air is removed. iii. Place a clean Sweeney filter holder loaded with a NEW Millipore filter on the end of the syringe. iv. Label the syringe: "Reynolds Lead Citrate, Date and initials ". v. Place the loaded syringe in the charged desiccator.
David Joswiak wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 16:55:28 2005
After receiving several off line comments from my friends about the fact that Graciela Matrajt was working with meteorites not biological samples and the staining methods I suggested wouldn't work, just want to add that I responded to what I thought was a general question i.e. "And I also wonder if there is a way/procedure of staining directly the TEM grids containing microtomed sections" In retrospect, he perhaps meant microtomed meteorite samples and not just any sample, in which case my response should be ignored. Sorry for the wasted words on the listserver. It is hard to get good help anymore :)
General thoughts on meteorite sections and staining: I have cut all sorts of materials i.e. metals, ceramics, ICs, polymers, catalysts as well as biological samples, but never meteorites. Not withstanding, if thin sections could be cut of meteorites, they may have enough different inclusions etc. to form their own contrast by atomic number and/or density difference. If it were more of a homogeneous material, then perhaps the same stains we use for polymers might work e.g. ruthenium or osmium tetroxide. Linda Sawyer published a book on Microscopy of Polymers which was updated in recent years. I would think that getting nice thin sections of meteorites might be more of a challenge than staining them.
Cheers Judy Murphy
Judy Murphy wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi, } I do not know of a stain that would differentiate organic carbon from } not-organic carbon. But am responding to the comment } } "And I also wonder if there is a } way/procedure of staining directly the TEM grids containing microtomed } sections on them." } } Most of the standard TEM section stains attach to proteins, nucleic } acids, or unsaturated lipids or to the Os which reduced those } unsaturated bonds. There are several specialty stains as well which } can attach to for instance, certain sugars. } However the standard staining method for TEM sections is usually the } Reynolds uranyl acetate, lead citrate stain or some derivative of } that. Lead citrate can be made from components (lead nitrate, sodium } citrate) or using the compound lead citrate directly (e.g. Venables } instant lead citrate stain). Generally I have found that making lead } citrate from its components is more stable for a longer amount of time } than using the "instant" lead citrate stains, however I have used both } successfully. When I use the "instant" lead citrate, I usually do so } by making it up fresh each time. } } Reynolds, E.S., 1963. The use of lead citrate at high pH as an } electron-opaque stain in electron microscopy, J. Cell Biol. 17, 208. } Venable, J. H. and Coggeshall, R., 1965. A simplified lead citrate } stain for use in electron microscopy, J. Cell Biol 25, 407. } General reference } Lewis, P.R., and Knight, D.P., 1977. Staining methods for sectioned } material, in "Practical Methods in Electron Microscopy", Volume 5, } Part I,Audrey Glauert (Ed), Elsevier/North-Holland Press. } } Below is one way to do the Reynolds uranyl acetate, lead citrate } stain. It is likely that the formatting will not be retained in the } text mode so in addition I am sending you off line a word document as } an attachment which will retain the format. The below method } indicates keeping the stains in syringes in a charged desiccator } however you can also keep the stains in the refrigerator, just so they } are sealed in something that keeps especially the lead citrate away } from air and UA away from light. } } Especially if you are interested in trying to find something that will } differentiate a very minute difference, it is suggested to use } deionized, glass distilled water. I have found when working with a } variety os polymer staining methods that water that is purified by a } resin column method (e.g. millipore) tends to bring along some of the } resin on occasion which also nicely stains. } } If you find such a stain that can differentiate what you are looking } for, would appreciate knowing about it. } } Thanks } Judy } } Judy Murphy, PhD } Microscopy, Imaging & Lab Design Consultant } Stockton, CA 95219 } murphyjudy-at-comcast.net } } Uranyl Acetate and Reynolds Lead Citrate Staining of Thin Sections for } TEM: } Preparation and Use } Judy Murphy 050124 } } Staining of TEM thin sections involves a two step process, first } staining with uranyl acetate and then with lead citrate. During the } lead citrate staining, it is imperative to minimize the exposure of } lead citrate with air to prevent precipitation on the sections of lead } carbonate. To this end, NaOH pellets are used which removes the CO2 } from the air. The more care that is taken to do the staining, the } cleaner the sections will look. } NOTES: • Preparation of Reynolds Lead Citrate stain takes } approximately 30-40 minutes. • Uranyl Acetate stain should be } stored in a brown bottle to prevent reaction with light. • Both } stock stains are stored in the refrigerator at 4OC. • Stains MUST } be Millipore filtered just before use. } • Small amounts of the stains may be stored in 3cc syringes with } the filters attached and placed in a charged desiccator. } 1. Boil deionized, distilled water as follow: } NOTE: Freshly boiled water must be used to make the 0.02 N NaOH } (if none available), water wash in wash bottle after staining, and to } fill 10 ml water beakers at least 2 times. If new Lead Citrate stain } needs to be made then it also needs to be made with freshly boiled water. } A. Prepare Beaker as follows: } a. Procure a clean 400-500 ml glass beaker. } CAUTION: Use a beaker 2-3 times larger than the volume of } water to be boiled } b. Rinse the clean beaker with a small amount of deionized, } distilled water. } B. Fill the rinsed beaker half full with deionized, distilled } water. } C. Place beaker in middle of microwave and close the door. } E. Microwave water for 10 mins. Exact time depends on the } microwave. NOTE: Water should be allowed to boil about 5 } minutes after it comes to a boil, which is about 10 min. in the } microwave oven. } CAUTION: Do not leave boiling water unattended as water may } boil over. } F. Using hot pad to protect skin, remove the beaker from the } microwave. } G. Cover the beaker with half of a petri dish. } H. Allow the water to cool to room temperature. } NOTE: It is important that as little CO2 which is found in } the air be allowed to enter back into the water. For that reason, } water should be boiled shortly before use, leaving enough time only to } cool the water. } } 2. Assemble stains and staining materials while waiting for water: } A. Freshly boiled and cooled deionized, distilled water (See } Step 1 for preparation). } B. 2% Uranyl Acetate (aqueous) stain in labeled, clean syringe } with FRESH Millipore filter attached (See Step 5A for preparation). } This is stored in a charged desiccator specifically for the stains. } C. Reynold's Lead Citrate stain in labeled, clean syringe with } FRESH Millipore filter attached (See Step 5B for preparation). This is } stored in a charged desiccator specifically for the stains. } D. NaOH pellets in sealed container. } E. Wash bottle containing 0.02M NaOH (for rinse after Lead } Citrate staining) } a. Measure out 180ml of the freshly boiled and cooled } deionized, distilled water. } b. Pour water into a clean and rinsed wash bottle. } c. Add 2 NaOH pellets. } NOTE: Each pellet weighs approximately 0.07g } d. Mix solution well but do not cause CO2 to enter. } F. Wash bottle containing freshly cooled boiled distilled water. } G. Assemble the following necessary materials on a lined tray: } a. Three large glass Petri dishes (tops and bottoms). Line } one Petri dish with a piece of filter paper indicated below. } b. One small Petri dish. } c. Three 10ml beakers. } d. Two waste beakers, one for the 0.02 N NaOH rinse waste } and one for the water rinse waste. } e. Two squares of Parafilm cut to fit into one of the large } Petri dish. } f. 2 pieces of filter paper, one for lining the Petri dish. } g. Filter paper arrows (fairly small arrows). } h. Clean EM tweezers, preferably locking tweezers, if } available (may use small o-ring to lock regular tweezers). } i. Grid box containing sections to be stained with proper } identification sheets. } } 3. Uranyl Acetate Staining: } A. Put grids that are to be stained within 20 min. in a covered } Petri dish lined with a piece of filter paper. } B. Heat grids by putting them in the 52°C over for 15 minutes. } A. Fill the 10ml beakers 3/4 full with boiled deionized, } distilled water. } B. Prepare Staining Petri dish as follows: } a. Place a square of parafilm in the bottom half of a glass } petri dish. } CAUTION: Be sure that parafilm is flat. } b. Place drops of 2% UA on the paraflim layer in the petri } dish, one per grid to be stained. } NOTE: Drops should be just larger than the grids. } C. Float each of the grids, singly, section-side down on a } separate drop of stain and cover the Petri Dish. } D. Allow grids to stain for approximately 25-30 minutes. } E. Remove the grids one at a time and immediately rinse by quickly } dipping 20 times into each of the three10ml beakers of fresh } deionized, distilled water. NOTE: Use a straight up and down } motion. } F. Holding the grid in the tweezers, remove the water retained on } the grid by touching filter paper arrows to the edge of the grid. } G. Place the grid on filter paper in a Petri dish. } H. Clean up of UA: } a. Dispose of stain in appropriate waste bottle under the } fume hood. } b. Dispose of solid waste (filter paper arrows, parafilm } etc.) in appropriate bio-hazardous waste container. } } 4. Lead Citrate Staining: } A. Fill the 10ml beakers approximately to 3 mm below the top, } with freshly boiled and cooled deionized, distilled water. } B. Prepare Petri dishes as follows: } NOTE: Use a double Petri dish system i.e. small Petri dish } top inside top and bottom of larger Petri dish. } a. Place a square of Parafilm in the bottom half of a glass } Petri dish. } CAUTION: Be sure that parafilm is flat. } b. Place 4 NaOH pellets close to but NOT touching where the } Lead Citrate droplets will be placed. } c. Place drops of Lead Citrate stain on the Parafilm layer } in an area that can be covered by the small Petri dish, one per grid } to be stained, near where the NaOH pellets are but NOT touching the } pellets. } NOTE: Drops should be just larger than the grids. } d. Immediately put the small Petri dish top over the } Parafilm area which has the Lead Citrate droplets and the NaOH } pellets. Cover also the large Petri dish. e. Allow it to } sit about 5 min. to remove CO2 from the atmosphere surrounding the stain. } C. When ready to stain, quickly open the Petri dishes and float } each of the grids, singly, section-side down on a separate drop of } stain and quickly cover the Petri Dish. } NOTE: Open the Petri dishes only as far as necessary to } minimize the amount of air (which contains CO2) that gets into the } Petri dishes. } D. Allow grids to stain for appropriate time. This can vary } between 5 to 20 min. } NOTE: Stain time will vary. Perform test for type of tissue } and stain. } Staining Time Test: Start with 3 grids that all contain } sections of the same thickness. These should be silver when viewed in } the ultramicrotome. Stain one grid for 5 min, one for 10 min, and one } for 15 min. with Lead Citrate. Uranyl acetate stain can all be 30 } min. each. E. Rinse the grids one at a time as follows: } a. Pick up the first grid and holding the grid with clean } tweezers, gently run a stream of 0.02M NaOH (in wash bottle) over the } grid and into a waste beaker. } NOTE: Start the stream of NaOH first and then bring the } grid into the stream so it doesn't put extensive washing force on the } grid. } b. Quickly, rinse the grid in the same manner with freshly } boiled deionized, distilled water in a wash bottle. } c. Immediately rinse the grid by quickly dipping at least } 20 times into each of the three 10ml beakers of fresh deionized, } distilled distilled water. } NOTE: Use a straight up and down motion. } F. Remove liquid on tweezers and grid as follows:] } a. Holding the grid in the tweezers, remove the water } retained in the tweezers by gently putting a paper arrow in between } the tweezers coming from the non-pointed end downward. } NOTE: Anti-capillary tweezers do not require this step. } b. Still holding the grid in the tweezers, remove the water } retained on the grid by carefully touching filter paper arrows to the } edge of the grid. } G. Place the grid on filter paper in a covered Petri dish to } dry. After a few minutes, the grid can be placed in a grid box } indicating proper labeling information on the Grid Box Information Sheet. } H. Be sure to clean tweezers with clean water before picking up } the next grid to be stained or before putting the tweezers away. } I. Clean up Lead Citrate waste as follows: } a. Dispose of used stain in appropriate waste bottle under } the fume hood. } b. Dispose of solid waste (filter paper arrows, Parafilm } etc.) in appropriate biohazardous waste container. } } 5. Preparation of Stains: } CAUTION: Wear Gloves when preparing stains. } A. 2% aqueous Uranyl Acetate Stain: } a. Measure 48ml deionized, distilled water into a } clean100ml beaker. } b. Weigh out 1 gram Uranyl Acetate. } c. Add measured Uranyl Acetate to the deionized, distilled } water in the beaker. } d. Bring volume up to 50ml with deionized, distilled water. } e. Stir solution on the magnetic stirrer to dissolve crystals. } f. Store stock stain in properly labeled brown bottle. } g. Prepare 3cc syringe of UA as follows: } i. Using a 3cc syringe, draw about 2ml of UA. } ii. Remove air from the syringe by holding the syringe } straight up (needle on top) and slowly push on the plunger until the } air is removed. } iii. Place a clean Sweeney filter holder loaded with a NEW } Millipore filter on the end of the syringe. } iv. Label the syringe: "2% UA, Date and initials ". } v. Wrap the syringe with tin foil to protect the stain from } light. } B. Reynold's Lead Citrate } CAUTION: Lead stains precipitate as white opaque carbonate } grains on grids if any contact with C02 is made. Be careful not to } breathe on stain drops or on grids while rinsing. } NOTE: Concentrated NaOH solutions should NOT be stored in } glass-stoppered bottles or the stoppers will "freeze" and not be able } to be removed. } a. Pour 30ml of the boiled and cooled deionized, distilled } water into a 50ml volumetric flask. } b. Weigh out the following: } i. Lead Nitrate crystals: 1.33 g } ii. Sodium Citrate crystals: 1.76 g } c. Add the Lead Nitrate and Sodium Citrate crystals to the } 30ml of water in the volumetric flask. } d. Mix the solution as follows: } i. Tightly seal the top of the volumetric flask with } parafilm. } ii. Shake vigorously for 1 minute. } NOTE: Solution should be milky white. } iii. Allow to stand with occasional mixing for 30 minutes. } e. Add 1 N NaOH until cloudiness disappears as follows: } i. Add one drop of 1 N NaOH. } ii. Swirl solution until mixed. } iii. Repeat until cloudiness disappears. } g. pH solution to pH 12 using 10 N NaOH and bring the } volume up to 50 ml with freshly boiled and cooled deionized, distilled } water. } CAUTION: pH must be maintained above PH 12 or carbonate } precipitate will form. } h. Store stain is stored in a tightly stoppered bottle. } i. Prepare 3cc syringe of Lead Citrate as follows: } i. Using a 3cc syringe, draw about 2ml of Lead Citrate. } ii. Remove air from the syringe by holding the syringe } straight up (needle on top) and slowly push on the plunger until the } air is removed. } iii. Place a clean Sweeney filter holder loaded with a NEW } Millipore filter on the end of the syringe. } iv. Label the syringe: "Reynolds Lead Citrate, Date and } initials ". } v. Place the loaded syringe in the charged desiccator. } } } } } } } } } } David Joswiak wrote: } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } Below is a question from a colleague (who is not on the list) regarding } } staining of organic carbon. } } } } Dave Joswiak } } University of Washington } } } } } } Hi, } } } } I would be interested in any information regarding staining of organic } } carbon prior to observing the sample in a TEM. I work with meteorites } } and I am trying to distinguish the organic carbonaceous phases from the } } not-organic carbon present in the samples. } } I wonder if there exists any staining method to distinguish the organic } } carbon from the rest(it does not need to be specific to a type of } } organic } } carbon such as proteins, etc.). And I also wonder if there is a } } way/procedure of staining directly the TEM grids containing microtomed } } sections on them. } } } } Thanks in advance. } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 17:34:35 2005
} Could anyone tell me how to measure and calculate the brightness and } dose on the fluorescent screen of a transmission electron microscope? } We have the Tecnai TEM. } Dear Lu, There are several ways to do this, but for the most accurate results, the screen should be calibrated with a suitable Faraday cup. When we first measured screen current with the utility in the low dose set-up, it did not agree with the measurement from the screen current utility--you may or may not have access to that utility, but your FEI service person does. Subsequently, we calibrated both measurements, and we now can be confident that either will give us a true reading. We have continued to check the CCD sensitivity, and, since that has not changed measurably, we can now use the CCD to determine the desired dose by setting the beam parameters to give us the number of counts per channel that corresponds to the number of electrons per nm^2. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 18:36:54 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (armstrong30-at-llnl.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, January 26, 2005 at 13:27:33 ---------------------------------------------------------------------------
Email: armstrong30-at-llnl.gov Name: Mike Armstrong
Organization: Lawrence Livermore Nat'l Labs
Title-Subject: [Microscopy] [Filtered] EM cooling stage questions and shopping (buyer)
Question: Hi!
I am interested in doing TEM on frozen (but not necessarily cooled to liquid nitrogen temperatures) samples in a JEOL 2000FX TEM. I have two sets of questions:
1) Does anybody out there have a cooling stage for this microscope that I could beg/borrow/rent/buy? Even if you have a lead that I could check out, it would be great if you could let me know.
2) Does anyone know of a Peltier cooled stage for TEM (for this microscope) that's a reasonable price? How hard do you think it would be to build such a thing? I only need to go to sub-freezing, not super cold, and it seems like this might be a cheaper alternative.
I'm new to EM, so if you have any other suggestions, they would be helpful. I'm kind of scraping to get this for as little as possible. If anybody can help me, I'd really appreciate it. This is for a high risk/payoff experiment and you will certainly get a mention (at least) in some acknowledgements if you can help me out.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sschul-at-vet.ksu.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, January 26, 2005 at 15:01:11 ---------------------------------------------------------------------------
Question: I am going to do a project that involves ticks. The ticks will feed on the dogs and then drop off, once they drop off and mature to adults I would like to do SEM and TEM on them to determine the location of the bacteria with in the tick itself. I believe that I will do critical point dry. I was wondering if you could help me out with a protocol for how to specifically fix the ticks to ensure that they are being processed correctly. Thanks
Hard to believe we are at Year 10 already. Thank you all for your support over the years.
As the list is now enforcing the One-Announcement-per-Course rule, don't expect further announcements.
Up-to-date information can always be found at the Course WWW site: www.3dcourse.ubc.ca
Cheers,
Jim P.
_______________________________
Tenth Annual INTERNATIONAL 12-Day Short Course on 3D Microscopy of Living Cells, June 11 - 23, 2005 (Pre-course: June 11)
Ninth, Post-course Workshop on 3D Image Processing, June 25 -27, 2005
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with the Departments of Pharmacology and Physiology and the Brain Research Centre, University of British Columbia, Vancouver, BC, Canada
DATES
Applications must be received by Tuesday, March 15, 2005 Deposit due Friday, April 15, 2005 Registration 5:00 - 7:00 PM Saturday, June 11, 2005 First Lecture 7:30 PM Saturday, June 11, 2005 Live-cell Course ends, noon Thursday, June 23, 2005 3D Image Processing Course, Saturday, June 25 - Monday, 27, 2005
APPLICATIONS DUE BY MARCH 15, 2005
APPLICATIONS Applicants must complete a questionnaire on the web to assess knowledge level, field of interest and proposed personal, live-cell, project. Enrollment will be limited to about 32 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms requesting information on field of interest and level of experience may be down-loaded from the WWW site at
www.3dcourse.ubc.ca/application.htm , or obtained from:
Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 250 N. Mills Street, Madison, WI, 53706 JBPAWLEY-at-WISC.EDU
Additional information is available from: www.3dcourse.ubc.ca/brochure.htm and links.
We expect to have at least 11, 3D microscope workstations for student use and there will be an international faculty of 20.
Application deadlines:
Application forms must be received for screening by March 15, 2005. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2005. In general, refunds of the deposit will only be possible if someone on the waiting list can take the place but this has not been a problem in previous years. The remaining balance is due before Registration.
Pre-Course Tuition (1/2 day Basic Optical principles) $100 (US) 3D Live-cell Course Tuition (includes lunches, snacks, 3 dinners): $2,650 (US) Workshop Tuition (includes lunches and snacks): $1,000 (US)
Room/board about $40/day (US) depending on room type.
I hope that this includes all of the information that you need, but if not, please get back to me.
-- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 19:24:11 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sdublin-at-emory.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, January 26, 2005 at 15:57:22 ---------------------------------------------------------------------------
Email: sdublin-at-emory.edu Name: Steven Dublin
Organization: Emory University
Education: Graduate College
Location: Atlanta, GA
Question: I am a grad student trying to make my own carbon coated grids. I have no problem depositing a carbon film on a glass slide and floating the film on water. Next,I drop the grids on the film. My problem occurs when I swipe the grids off the surface of the water with a glass slide. The residual water on the slide wrinkles and tears the film, so I am left with an uneven, broken film across the surface of the grid.
Does anyone have advice on how to get the grids out of the water without tearing the film? What about grid glue? Is this required?
On Jan 26, 2005, at 4:42 PM, by way of MicroscopyListserver wrote:
} I am interested in doing TEM on frozen (but not necessarily cooled to } liquid nitrogen temperatures) samples in a JEOL 2000FX TEM. I have two } sets of questions: } } 1) Does anybody out there have a cooling stage for this microscope } that I could beg/borrow/rent/buy? Even if you have a lead that I could } check out, it would be great if you could let me know. } } 2) Does anyone know of a Peltier cooled stage for TEM (for this } microscope) that's a reasonable price? How hard do you think it would } be to build such a thing? I only need to go to sub-freezing, not super } cold, and it seems like this might be a cheaper alternative. } } I'm new to EM, so if you have any other suggestions, they would be } helpful. I'm kind of scraping to get this for as little as possible. } If anybody can help me, I'd really appreciate it. This is for a high } risk/payoff experiment and you will certainly get a mention (at least) } in some acknowledgements if you can help me out. } Dear Mike, I can't help you regarding the JEOL 2000FX, but an additional possibility for a cooled stage is to beg/borrow/rent/buy a LN2 stage and fill it with a dry ice-ethanol slurry (which a JEOL rep can tell you if this mixture could cause damage). Ice-salt-water can also be used for barely sub-freezing temps, but I would rinse thoroughly afterward. If you want to build a Peltier-cooled stage, you just need the appropriate thermal contact to which one can attach the Peltiers; however, that might be easier said than machined. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 20:48:30 2005
Instead of picking up the carbon from the water surface with a glass slide all at once, it may be possible to break up the carbon on the water surface slightly and pick up a small 3 mm piece individually with each grid from underneath.
Usually for carboned only grids for virus examination, we would evaporate carbon onto freshly cleaved mica pieces (available from EM suppliers). We then floated the carbon films onto the surface of the water. Using freshly acetone cleaned grids, we put one grid at a time underneath the pieces of carbon and picked it up. Freshly cleaning them, would lower the surface tension when the grid went into the water. We generally used 400 mesh Cu grids to support the films.
"Does anyone have advice on how to get the grids out of the water without tearing the film? What about grid glue? Is this required? "
I have used grid glue routinely for picking up sections on naked grids but have never found it necessary for carbon only grids.
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From MicroscopyL-request-at-ns.microscopy.com Wed Jan 26 21:20:57 2005
Dear Steven Try to use piece of Parafilm (instead glass) to pick up the grids. Parafilm should be larger than carbon film. You need completely immerse Parafilm into the water and than remove it quickly. Grids should be clean to stick to the carbon. Depends from carbon thickness, you need to use grids of corresponding mesh: As thinner carbon, higher mesh should be. Carbon, which is slightly brownish (so you could see carbon by naked eye) may be mounted on 400-mesh grids. The alternative (there are so many ways) way is to put plastic film on the grids (which is easy) first, then evaporate carbon and then dissolve plastic (it would "glue" carbon to the grid). You may also leave plastic on the grid. Generally, carbon do not stick well to the naked grids, so people used plastic support film or "holey film". You may also "pre-treat" grids with plastic (a drop of very diluted plastic solution per grid, dry before use) before mounting the carbon. I hope it helps. Have a great day, Sergey
At 05:29 PM 1/26/2005, 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 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
It has been a long time since I've made carbon coated grids, but one of the approaches we used was to wrap a piece of coarse copper mesh around some wire (coat hanger would do) to create a support that we could place under water. We then dunked the grids through the water onto this support, and then floated off the carbon film onto the surface of the water. We were then able to manipulate the grid holder under the carbon film, and pick up the whole thing through the water.
The simple method, of course, was to make thin formvar coated grids, and evaporate some carbon on to them. Does your work preclude this approach?
Joel
Date sent: Wed, 26 Jan 2005 18:53:21 -0800 } From: Judy Murphy {murphyjudy-at-comcast.net} To: by way of Ask-A-Microscopist {sdublin-at-emory.edu} Copies to: microscopy-at-microscopy.com
Hi all,
I've been trying to calibrate the objectives of a brightfield microscope from 2,5x to 100x to its best, but still I have an average deviation of 3,9% if I compare the same object measured on all magnifications. So if I measure something with the 2,5x, it appears longer then with the 40x objective. Is 3,9% acceptable as deviation and if not, what is? Is there a general rule or a paper about this? Thanks in advance,
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 27 10:01:57 2005
Announcing a 5 day intensive AFM short course entitled "AFM and Other Scanned Probe Microscopies". It is being taught at N.C. State University in Raleigh, NC by Prof. Phil Russell, Dr. Joe Griffith and others from June 13 -17, 2005.
This one-week short course on atomic force microscopy has evolved from the numerous Scanned Probe Microscopy courses developed and taught by Prof. Russell over the past 2 decades. It is designed for technicians, scientists, engineers and researchers. The course includes lectures and laboratories with hands-on time using a variety of scanning probe microscope (SPM) systems.
For more information go to www.ncsu.edu/aif/afmcourse
I am looking to obtain some references pertaining to the analysis of human hair, both morphologic and spectroscopic. I'm trying to get my feet wet and need a place to start. Does anyone have particular recommendations?
Any help is greatly appreciated.
Thanks in advance!
George R. Munzing Jr. Engelhard Corporation Strategic Technologies Group Iselin, NJ 08830 TEL: 732-205-7030 FAX: 732-494-3283 e-mail: george.munzing-at-engelhard.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 27 17:32:53 2005
Sent: Thursday, January 27, 2005 4:20 PM To: Microscopy-at-MSA.Microscopy.Com
Hello All,
I am looking to obtain some references pertaining to the analysis of human hair, both morphologic and spectroscopic. I'm trying to get my feet wet and need a place to start. Does anyone have particular recommendations?
Any help is greatly appreciated.
Thanks in advance!
George R. Munzing Jr. Engelhard Corporation Strategic Technologies Group Iselin, NJ 08830 TEL: 732-205-7030 FAX: 732-494-3283 e-mail: george.munzing-at-engelhard.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 27 18:01:07 2005
Been dabbling in forensic microscopy for the last 5 yrs with a high school forensics program and have been using the following for microscopy of human hair
Atlas of Human Hair: Microscopic Characteristics ISBN: 0-8493-8134-7
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From MicroscopyL-request-at-ns.microscopy.com Thu Jan 27 19:21:21 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gunkelrl-at-slu.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 27, 2005 at 12:06:03 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kaprelyants-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, January 27, 2005 at 15:24:07 ---------------------------------------------------------------------------
Email: kaprelyants-at-yahoo.com Name: Alex Kaprelyants, Ph.D.
Hey Judy I have this book. I think George should be warned though that while the book is useful, it is only about a quarter of an inch thick and small, and cost about $107 Canadian for 83 pages. It was a shock when I got it.. I expected at least a tome of some sort. Elaine
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-- Dr. Elaine Humphrey Director, BioImaging Facility 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-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 27 20:33:32 2005
A couple to start you off with.... There may be more in a day or so when I get a chance to go through some more....
Jones LN. Clinics in Dermatology, 2001;19:95-103.
Rogers GE. Electron microscope studies of hair and wool, Ann NY Acad Sci 1959;83:469-485.
Hallegot P, Corcuff P. High-spatial-resolution maps of sulphur from human hair sections: An EELS study. J. Micrsc 1993;172:131-136.
Jones LN, Cholewa M, Kaplin IJ, et al. Elemental distributions in keratin/follicle sections. Proc Eighth Int Wool Textile Conf. Christchurch NZ 1990;1:246-255.
Rogers GE, Reis PL, et al., editors. The biology of wool and hair. London: Chapam and Hall, 1989.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 27 22:18:57 2005
is this evian or naive of me, but have you tried med line? maybe pubmed. john --- George_Munzing-at-engelhard.com wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello All, } } I am looking to obtain some references pertaining to } the analysis of human } hair, both morphologic and spectroscopic. I'm trying } to get my feet wet and } need a place to start. Does anyone have particular } recommendations? } } Any help is greatly appreciated. } } Thanks in advance! } } George R. Munzing Jr. } Engelhard Corporation } Strategic Technologies Group } Iselin, NJ 08830 } TEL: 732-205-7030 } FAX: 732-494-3283 } e-mail: george.munzing-at-engelhard.com } } }
__________________________________ Do you Yahoo!? All your favorites on one personal page – Try My Yahoo! http://my.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 28 01:38:02 2005
Variations on a goggle search www.google.com "human hair" preparation microscope look promising try replacing perpareion with other terms like microtome, embedding, spurs, resin, etc and you should find a lot of things.
On McCrone's Modern Microscopy publication http://www.modernmicroscopy.com/main.asp A Microscopical Study of Exotic Animal Hair: Part 1 by Kristen D. Skraba, McCrone Associates, Westmont, IL. Might be of interstest even thought it is about animal hair it shows a few tecniques and methods. The biligoriphy should help you some.
Check your local library if it is not a major reserch universty it is unlikely they will have anything but you can search the Libary of Congress at: http://catalog.loc.gov/cgi-bin/Pwebrecon.cgi?DB=local&PAGE=First and use inter library loan to get anything that intrests you. It's not the instant access that the web give but it a geat service partiulary for obscure papers from all over the world.
Good luck Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
} } Hello All, } } } } I am looking to obtain some references pertaining to } } the analysis of human } } hair, both morphologic and spectroscopic. I'm trying } } to get my feet wet and } } need a place to start. Does anyone have particular } } recommendations? } } } } Any help is greatly appreciated. } } } } Thanks in advance! } } } } George R. Munzing Jr. } } Engelhard Corporation } } Strategic Technologies Group } } Iselin, NJ 08830 } } TEL: 732-205-7030 } } FAX: 732-494-3283 } } e-mail: george.munzing-at-engelhard.com } } } } } } } } } } } } __________________________________ } Do you Yahoo!? } All your favorites on one personal page – Try My Yahoo! } http://my.yahoo.com } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 28 07:21:32 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spagosullamiavaligia-at-libero.it) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, January 28, 2005 at 05:35:21 ---------------------------------------------------------------------------
Email: spagosullamiavaligia-at-libero.it Name: Marco Cattaneo
Organization: University of Cosenza
Education: Graduate College
Location: Cosenza, Italy
Question: Dear Sir/Madam, i need to keep cells alive under the microscope for several hours. I realize that exist two microscope incubation systems: the box surrounding the microscope and the chamber fitting on the microscope stage. Could you please tell me which are the meaningful differences between these two microscope incubation methods? Is it true that the chamber fitting on the stage causes a focus drift? Which are the most reliable brand for these systems (obviously with the best quality/price ratio)?
Is there an automated cryotome that can face multiple samples with little human intervention?
Jerry W. Ball Advanced Characterization Product Technology BTEC West Complex ExxonMobil Chemical Company 5200 Bayway Dr. 77520-2101 Phone: 281-834-1718 Fax: 281-834-1793 E-Mail: jerry.w.ball-at-exxonmobil.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 28 11:29:44 2005
Dear Listers I have a researcher who would like to create cross sections of 2 - 3 year old pine sapling stems. The sections do not need to be terribly thin (50 - 100 micrometers) but he would like to have the surface as smooth as possible without embedding the tissue (he is a physiologist interested in chloroplasts). My background is with animal tissue so I'm at a bit of a loss in how to advise him. Any suggestions would be gratefully accepted. Thanks in advance...
Richard Harris Laboratory Supervisor Microscopy, Imaging and Analysis Department of Biology University of Western Ontario London Ontario CANADA N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 28 14:07:23 2005
Richard, If you have access to a vibratome than use that to cut the sections. Also, with some practice you may be able to cut free-hand sections that are thin enough for your purposes.
Alternatively you can buy little hand microtomes from major science or school supply companies. These cost about $100 I think and have a sample chuck that is raised by a little micrometer screw assembly through a flat platform. You insert your stem and then gradually raise it in height. Then you slide a long razor blade across the piece using the platform as a support. It is really a simple but very useful device.
If we have a tissue that is too soft or small to hold well in the chuck then we just hollow out a piece of carrot and place the twig, etc inside. This will clamp well and hold the tissue well to get minimal crushing and distortion during cutting.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 1/28/05 12:33 PM, "Richard Harris" {rjharris-at-uwo.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Listers } I have a researcher who would like to create cross sections of 2 - 3 year } old pine sapling stems. } The sections do not need to be terribly thin (50 - 100 micrometers) but he } would like to have the surface as smooth as possible without embedding the } tissue (he is a physiologist interested in chloroplasts). My background is } with animal tissue so I'm at a bit of a loss in how to advise him. Any } suggestions would be gratefully accepted. } Thanks in advance... } } Richard Harris } Laboratory Supervisor } Microscopy, Imaging and Analysis } Department of Biology } University of Western Ontario } London Ontario CANADA } N6A 5B7 } Ph. 519-661-2111 ext. 86780 } Fax 519-661-3935 } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 28 17:10:31 2005
I would recommend a sliding microtome (aka sledge microtome) if you can find one. The other possibilities are a cryostat or hand sections made with a double-edged razor blade. The hand sections are OK if he doesn't need consistency. Kim
{} {} {} {} {} {} {} Kim Rensing PhD Bio-Imaging Facility, UBC 6270 University Boulevard Vancouver, BC V6T 1Z4 {} {} {} {} {} {} {}
On Friday, January 28, 2005, at 09:33 AM, Richard Harris wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Dear Listers } I have a researcher who would like to create cross sections of 2 - 3 } year } old pine sapling stems. } The sections do not need to be terribly thin (50 - 100 micrometers) } but he } would like to have the surface as smooth as possible without embedding } the } tissue (he is a physiologist interested in chloroplasts). My } background is } with animal tissue so I'm at a bit of a loss in how to advise him. Any } suggestions would be gratefully accepted. } Thanks in advance... } } Richard Harris }
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 28 21:59:09 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (svanhorn-at-notes.cc.sunysb.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, January 28, 2005 at 14:23:09 ---------------------------------------------------------------------------
Email: svanhorn-at-notes.cc.sunysb.edu Name: Sue Van Horn
A good sharp wood plane iron or wide wood chisel sharpened as you would a microtome knife are much better than the straight razor that usually comes with them. Cementing a glass microscope slide on each side of the hole in the face of the microtome will help too. Trim the glass slides so they don't cause a danger you the operator and use cutting edges made from good steel not the imported stuff. You may have to find and old chisel or plane to get a good piece of steel but the good modern brands should be good enough if you don't have and old on laying around.
Razor blades and straight razors are not stiff enough to good sections from soft tissue let alone hard stuff.
Try cutting with a slicing motion instead of with the blade at 90 degrees to the direction of travel.
Good luck Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
Debby Sherman wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Richard, } If you have access to a vibratome than use that to cut the sections. Also, } with some practice you may be able to cut free-hand sections that are thin } enough for your purposes. } } Alternatively you can buy little hand microtomes from major science or } school supply companies. These cost about $100 I think and have a sample } chuck that is raised by a little micrometer screw assembly through a flat } platform. You insert your stem and then gradually raise it in height. Then } you slide a long razor blade across the piece using the platform as a } support. It is really a simple but very useful device. } } If we have a tissue that is too soft or small to hold well in the chuck } then we just hollow out a piece of carrot and place the twig, etc inside. } This will clamp well and hold the tissue well to get minimal crushing and } distortion during cutting. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } On 1/28/05 12:33 PM, "Richard Harris" {rjharris-at-uwo.ca} wrote: } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 29 16:08:46 2005
Having cut a lot of wood and especially pine, maybe I can add a comment or two here. The requirement "smooth as possible" is a little vague; it would help to know how these sections are going to be examined i.e. optical, SEM, AFM?
1. From my experience, I've found the best way to section this material is using a regular microtome knife; disposable microtome blades just don't hold up well enough. A good well sharpened microtome knife will do a great job and stand up to the hard xylem elements.
2. Depending on the diameter of the sapling stems, they may have to be surrounded (not embedded) with something to make them more rigid. Options range from Paraplast to Epoxy depending on how hard the stem material is.
3. When sectioning, it helps to soak the block in ice water (or warm water depending on the material used in #2) to soften the woody portions of the stem.
4. Fixing Pinus tissue is a whole other problem!
Good luck on what can be a challenging project
Damian Neuberger
} Dear Listers } I have a researcher who would like to create cross sections of 2 - 3 year } old pine sapling stems. } The sections do not need to be terribly thin (50 - 100 micrometers) but he } would like to have the surface as smooth as possible without embedding the } tissue (he is a physiologist interested in chloroplasts). My background is } with animal tissue so I'm at a bit of a loss in how to advise him. Any } suggestions would be gratefully accepted. } Thanks in advance... } } Richard Harris } Laboratory Supervisor } Microscopy, Imaging and Analysis } Department of Biology } University of Western Ontario } London Ontario CANADA } N6A 5B7 } Ph. 519-661-2111 ext. 86780 } Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 30 16:00:29 2005
If your researcher wants to use fresh wood sections, the sliding or sledge microtome suggested by Kim Rensing will do a great job, as long as your knives are sharp. We found that the disposable blades are not stiff enough for doing this on the sledge microtome, you need to use the large metal blades. A colleague recently sectioned 2-year old poplar stems this way. We're lucky that a local histologist will sharpen the blades for us.
If you use a hand microtome, a non-flexible blade, something like a cut-throat razor, is probably necessary to get reasonably consistent sections. With hard tissue, the flexible blades chatter on the sledge microtome (on our one, anyway), and tend to flex and scoop out the middle of the tissue on a hand microtome.
For support material, we use high density foam, the type used for insulating refrigerators. We have a large slab of this and just cut a small piece approximately to shape to fit around the tissue, in the same way we get students to shape carrot pieces around tissue for hand sectioning. Everything from shoot apical meristems to cores of 50-year old trees have been sectioned this way on the sledge microtome.
Good luck! cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
} From: Richard Harris {rjharris-at-uwo.ca} } Date: Fri, 28 Jan 2005 12:33:31 -0500 } To: MSA Listserver {microscopy-at-MSA.microscopy.com} } Subject: [Microscopy] Sectioning woody plant stems } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Listers } I have a researcher who would like to create cross sections of 2 - 3 year } old pine sapling stems. } The sections do not need to be terribly thin (50 - 100 micrometers) but he } would like to have the surface as smooth as possible without embedding the } tissue (he is a physiologist interested in chloroplasts). My background is } with animal tissue so I'm at a bit of a loss in how to advise him. Any } suggestions would be gratefully accepted. } Thanks in advance... } } Richard Harris } Laboratory Supervisor } Microscopy, Imaging and Analysis } Department of Biology } University of Western Ontario } London Ontario CANADA } N6A 5B7 } Ph. 519-661-2111 ext. 86780 } Fax 519-661-3935 } }
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 30 16:28:12 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Jacob.blumenthal-at-weizmann.ac.il) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, January 30, 2005 at 04:03:29 ---------------------------------------------------------------------------
Email: Jacob.blumenthal-at-weizmann.ac.il Name: Jacob Blumenthal
Organization: weizmann institute of science
Title-Subject: [Microscopy] [Filtered] MListserver: negative staining of mRNA
Question: Hello, I would like to know if it possible to see mRNA molecule after the ribosomes came off of it using negative staining with uranyl acetate? It appears as a long thin string (not a straight one). Another question is what is diameter of a single ribosome as it apperas in em using negative staining? And if you can always see the forming peptides coming out from each ribosome? I am working on eukaryotic cells. Thanks,
Thank you all for your replies and suggestions. I think I have enough to keep me busy for quite a while.
Regards
George R. Munzing Jr. Engelhard Corporation Strategic Technologies Group Iselin, NJ 08830 TEL: 732-205-7030 FAX: 732-494-3283 e-mail: george.munzing-at-engelhard.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 31 14:05:10 2005
For the last year or so we have been using a Coolpix 4500 interfaced to a petrographic microscope, yielding considerable user satisfaction.
We have the WPI Adaptor Kit, P/N 501384, which has a 28mm male thread that screws directly into the filter thread on the end of the lens of the Coolpix. Although the interfaceability of the Coolpix is a bit primitive, as all images are stored only on the card memory and have to be transferred to a computer either via the USB cable (slow) or by removing the card (fiddly), the mechanical coupling is firm and good, the optics work well.
The camera has now died, and I have to plan its replacement in case it turns out to be not repairable.
The Coolpix 4500 has been discontinued, and my local Nikon agents advise that there is no comparable Nikon replacement. As I understand it, the reasons for the widespread use of the 4500 for this application include the fact that it has a thread on the end of the lens for easy attachment of the adaptor, and that the end of the lens doesn't rotate or otherwise move as the camera changes its focus or zoom.
The WPI kit includes adaptors to step up to match camera female threads of 37mm and 43mm, although I can see that the more adaptors that are used, the less rigid the whole setup may become.
What alternative cameras are currently being successfully used out there?
Preferably with filter threads of 28, 37, or 43mm so that I can continue to use my WPI kit.
cheers and tia
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 31 14:28:31 2005
Does anyone have a manual for a Sorvall TC-2 tissue sectioner they could copy & send me?
On a completely different note, does anyone know how one could go about conducting a very brief (two or three multiple choice questions), non-commercial survey of career scientists? I'd like to make it so the responses are anonymous, thereby increasing likelihood of candid answers.
Paul :0)
---------------------------------------------------------------------- "Keep your eyes slightly wide and blank. Show no interest or excitement." - Dr. Miles Bennell Invasion of the Body Snatchers (1956)
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 31 15:59:26 2005
Dear Jackob It's possible to see the poly-ribosomes, when many individual ribosomes attached to single mRNA molecule. You may find many of articles already published on this matter. I don't remember did they use negative staining or shadowing, I would think: both. As far as I remember, the ribosomes looks like spherical particles without any details. The single ribosome visualization is another story. Yes, you could see easily individual ribosome by negative staining and size for 70S E.coli ribosome is about 2.6 nm (approx. diameter). Eucariotic ribosome may be a little bigger. If you are talking about mRNA attached to the ribosome - it's completely different story. You need to keep in mind that in order to function, ribosome needs at least two molecules of tRNA, mRNA and a bunch of "factors" - all this stuff attached to the ribosome making the picture very complicated. In addition, mRNA will never be like "linear structure" under physiological conditions: it would form compact structure, which is very difficult to made "straight"... synthesized peptide would act in similar way - you never will see peptide as a linear structure: it is immediately folded at the exit (even before). So, finally, you have the ribosomal particle surrounded with bunch of other molecules. All of them are sort of globular and interact with each other... There are million articles published on this matter. You may take a look on works by Joahim (spell?) Frank - he did 3D reconstruction of bacterial (?) ribosome with different factors, mRNA etc... Do google search on "ribosome structure Frank"... Good luck! Sergey
At 04:36 PM 1/30/2005 -0600, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 31 17:10:55 2005
I can't help on the microtome, but MME has been conducting surveys for 15+ years. Contact me off-line and we'll see what we can do.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 02:37 PM 1/31/2005, pgrover wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 31 18:30:39 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hgabrisc-at-uno.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, January 31, 2005 at 17:06:42 ---------------------------------------------------------------------------
Email: hgabrisc-at-uno.edu Name: Heike Gabrisch
Organization: Universit of New Orleans
Title-Subject: [Microscopy] [Filtered] MListserver: post doctoral position
Question: We have an immediate opening for a post-doctoral scholar. Her/his duties include the characterization of transition metal compounds by electron diffraction techniques. Transition metal compounds are used as intercalation compounds in rechargeable Li-ion batteries. The position requires hands-on experience in transmission microscopy - a background in crystallography is a plus. Contact : Heike Gabrisch (hgabrisc-at-uno.edu)
This has been a hot topic on the Yahoo microscope list. http://groups.yahoo.com/group/Microscope/
I have been told by a reliable source a CoolPix 5000 and even the 5400 can be use with UR-E6 for the 5000 and ER-U11 for the 5400 a part that cost less than 20 dollars in place of the CP 600-4500.
There is no other cameras that will work the optics the CoolPix 600-4500 I now of with out custom parts.
If the 4500 was satisfactory just buy a couple of used ones from some one with good reputation on ebay. They are pretty sturdy cameras. If you have problems with artifacts http://www.couger.com/microscope/shootout/shootout.html when you use the zoom function on the camera you might want to consider buying a CoolPix 990 or go the the 5,000 that doesn't have this problems.
Make sure if you buy a new camera you buy with the right to return it if doesn't work to suit you.
Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
Ritchie Sims wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Happy New Year } } For the last year or so we have been using a Coolpix 4500 interfaced to a petrographic } microscope, yielding considerable user satisfaction. } } We have the WPI Adaptor Kit, P/N 501384, which has a 28mm male thread that screws } directly into the filter thread on the end of the lens of the Coolpix. Although the } interfaceability of the Coolpix is a bit primitive, as all images are stored only on the card } memory and have to be transferred to a computer either via the USB cable (slow) or by } removing the card (fiddly), the mechanical coupling is firm and good, the optics work } well. } } The camera has now died, and I have to plan its replacement in case it turns out to be } not repairable. } } The Coolpix 4500 has been discontinued, and my local Nikon agents advise that there } is no comparable Nikon replacement. As I understand it, the reasons for the } widespread use of the 4500 for this application include the fact that it has a thread on } the end of the lens for easy attachment of the adaptor, and that the end of the lens } doesn't rotate or otherwise move as the camera changes its focus or zoom. } } The WPI kit includes adaptors to step up to match camera female threads of 37mm and } 43mm, although I can see that the more adaptors that are used, the less rigid the whole } setup may become. } } What alternative cameras are currently being successfully used out there? } } Preferably with filter threads of 28, 37, or 43mm so that I can continue to use my WPI } kit. } } } cheers and tia } } rtch } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 1 01:11:15 2005
BTW: the ribosome is about 25 to 30 nm in diameter, depending on the projection, but not 2.6 nm. Just a typing error, I assume. Kind regards, Reinhard ----------------- PD Dr. Reinhard Rachel Universitaet Regensburg Microbiology Universitaetsstr. 31 D-93053 Regensburg - Germany
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 1 08:04:32 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Somayyeh_kheiri-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 1, 2005 at 06:45:57 ---------------------------------------------------------------------------
Nextek Inc. has an immediate opening for the following failure/materials analysis engineer position. If interested, please fill out an application and send your resume to our human resources department (see our web page link at the end of the job posting).
Feel free to contact me if you have any questions. Thanks, Wade McFaddin Nextek Inc. 256-772-1995 ext.1064
Report directly to the Manager of Analytical Service or Vice President of Product Assurance.
Perform material failure analyses such as FTIR analysis, thermal analysis, scanning electron microscope (SEM) and energy dispersive spectroscopy (EDS) analysis, C-mode scanning acoustic microscopy analysis, cross section evaluation of printed circuit boards (PCB) and printed circuit board assemblies (PCBA). Support in-house PCBA manufacturing process and external customer requirements.
Evaluate final results of analysis, prepare and present reports outlining the outcome of analysis, and make recommendations for actions necessary to achieve desired results.
Job Requirements: Communication, presentation, and technical writing skills to effectively communicate to and from a diverse group such as the customer, Nextek employees and suppliers. Electronic manufacturing experience required.
Experience/Education/Training Requirements: B.S. degree in Engineering, Materials Science or Science or equivalent 5 - 10 years of electronic material laboratory experience
If you meet the requirements for this job and would like to be considered, please complete the
Nextek Employment Application on our website at www.nextekinc.com and submit to Human Resources.
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 1 13:58:53 2005
I have a colleague who is interested in finding SEM micrographs of mycoplasm (note: not mycoplasma!). We are trying to take pictures of it, but find ourselves confounded by apparent detritus in the growth media, such that images of the "fresh" media alone look very similar to the organisms that have been cultivated. If anyone has done work with this organism, and could provide us with one or two pictures to use as a reality check, it would be greatly appreciated!
Sincerely,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 1 14:29:47 2005
} Yes, I am sorry, it was a typo: 70S E.coli ribosome is about 26 nm in } coronal projection. Thanks Reinhard for correction!
} Sergey } } BTW: the ribosome is about 25 to 30 nm in diameter, } depending on the projection, but not 2.6 nm. } Just a typing error, I assume. } Kind regards, } Reinhard } ----------------- } PD Dr. Reinhard Rachel } Universitaet Regensburg } Microbiology } Universitaetsstr. 31 } D-93053 Regensburg - Germany
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 1 18:22:44 2005
I've been given some vanishingly small specimens to embed for TEM. They're peels from the surface of a retina - at most several cells thick and usually less than 1x2 mm in area. They are also made mostly of basement membrane, with a few whole cells and some cell fragments, so not only are they almost invisible to start with, but they remain invisible even after osmium. Pinning them down won't work - they're too small. I've embedded some once using a dissecting microscope, but it was a lot of work and even so I lost 2 of the 6. I thought of embedding first in agar, but then I'll end up with an invisible specimen somewhere in the resin, which will take longer find by serial sectioning than I'm prepared to spend.
Any ideas would be most welcome!
Diana
Diana van Driel Dept Ophthalmology Sydney University GPO Box 4337 Sydney, NSW AUSTRALIA 2001
I once had specimens so small the final pellet had an area of only one or two grid holes (cryosectioned too!) so I think I may be able to help here.
After lots of trials I eventually put the specimens in a thin polypropylene centrifuge tube (about 0.5ml total volume), re-suspended them in 10% gelatin (warm) and pelleted them down to the bottom of the tube. At the time the tubes had sharp pointy ends so the pellet of cell fragments went to the very tip if I used a horizontal rotor. I am not sure that these tubes are available any more.
Anyway, after the gelatin had gelled, I cut off the tip of the tube, which is fairly easy if the tube is made of polypropylene, and embedded it using a routine protocol. The gelatin turned dark and the tip was easily visible so I was able to orientate the block to cut the tip without problem.
Since then, I have been working with slightly larger amounts of cell fragments and have found that if you centrifuge the specimens down in warm gelatin in a regular Eppendorf tube, the pellet always deposits at the same place. I orientate myself in a fixed angle rotor (45 degrees) by putting the lid hinge outwards. I look for the pellet at the bottom of the tube under the hinge. Always wait for the gelatin to gel and cut out the pellet. Sometimes you cut where you think the pellet is. The smaller the gelatin block the easier you will be able to section.
If you were to be doing this for immunolocalization then agarose (2%) may be better. You can visualize the block after embedding by adding some color (alcian blue, dextran blue etc) to make it visible in the polymerized resin.
I know that some people say you shouldn't use gelatin because of its high affinity for water, but embedding protocols using gelatin have worked for me for many years.
Regards,
Paul Webster.
Paul Webster, Ph.D. Director Ahmanson Advanced EM and Imaging Center House Ear Institute 2100 West Third Street Los Angeles CA 90057-1922 Phone: 213 273 8026 Fax: 213 13 739 E-mail: pwebster-at-hei.org
On 2/1/05 4:20 PM, "Diana van Driel" {dianavd-at-eye.usyd.edu.au} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hi all, } } I've been given some vanishingly small specimens to embed for TEM. } They're peels from the surface of a retina - at most several cells } thick and usually less than 1x2 mm in area. They are also made mostly } of basement membrane, with a few whole cells and some cell fragments, } so not only are they almost invisible to start with, but they remain } invisible even after osmium. Pinning them down won't work - they're too } small. I've embedded some once using a dissecting microscope, but it } was a lot of work and even so I lost 2 of the 6. I thought of embedding } first in agar, but then I'll end up with an invisible specimen } somewhere in the resin, which will take longer find by serial } sectioning than I'm prepared to spend. } } Any ideas would be most welcome! } } Diana } } } } Diana van Driel } Dept Ophthalmology } Sydney University } GPO Box 4337 } Sydney, NSW } AUSTRALIA 2001 } } Phone 61 2 93827278 } Mobile 0423 151614 } FAX 61 2 93827318 } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 1 20:41:08 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 1, 2005 at 10:54:28 ---------------------------------------------------------------------------
Email: subramss-at-email.uc.edu Name: Jimble
Organization: University of Cincinnati
Title-Subject: [Microscopy] [Filtered] Phosphorus contamination in the ESEM
Question: Dear friends and elders,
I was wondering if someone would be able to help me track down a phosphorus contamination issue which I am observing in my ESEM when I use a hotstage. I am giving details of the instrument and conditions below. The materials I am observing are high purity oxides and should have but trace if any contamination of phosphorus.
Instrument: FEI XL-30 ESEM FEG with Diffusion pump backed a Mechanical pump. The system does not have any in-line filters or residual gas analyzers. Oils: Edwards Ultra 19 (Mechanical) Santovac 5 (Diffusion pump)
Phosphorus (POX)only appears on the MSDS of the Edwards oil as a combustion product but, vacuum gurus from Varian think that in order for mechanical pump backstreaming to occur pressures well below (100 millitorr) need to be present for pressure equilibriation and subsequent backstreaming. Even then with a 2 torr H2O pressure I expect the P traces should still be minimal preventing the contaminant from completely overtaking the process.
I would be greatful for an input as I am currently at a loss. I have attempted numerous methods (EDS, EBSD, XRD, XPS, TEM) in trying to get a clue to this problem and am yet to come up with any real conclusions.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Palash.Gangopadhyay-at-fys.kuleuven.ac.be) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, February 1, 2005 at 12:27:56 ---------------------------------------------------------------------------
Email: Palash.Gangopadhyay-at-fys.kuleuven.ac.be Name: Dr. Palash Gangopadhyay
Organization: Molecular and Nano Materials, K U Leuven
Education: Graduate College
Location: Leuven, Belgium
Question: Dear Microscopists
I will appreciate very much if somebody can direct me to an online AFM / MFM / STM course immediately available.
Hi Diana, Many years ago we often embedded spermatozoa which were also invisible. We got around it by staining them in methylene blue which gave them some color so we could find them in the capsule. As you did, we embedded them in a minimum of 2% agar which held them together while they were processed. Don't know if the MB would stain what you are looking at, but it might be worth a try or even perhaps some stain that might stick to what is in your sample if MB doesn't work.
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi all, } } I've been given some vanishingly small specimens to embed for TEM. } They're peels from the surface of a retina - at most several cells } thick and usually less than 1x2 mm in area. They are also made mostly } of basement membrane, with a few whole cells and some cell fragments, } so not only are they almost invisible to start with, but they remain } invisible even after osmium. Pinning them down won't work - they're } too small. I've embedded some once using a dissecting microscope, but } it was a lot of work and even so I lost 2 of the 6. I thought of } embedding first in agar, but then I'll end up with an invisible } specimen somewhere in the resin, which will take longer find by serial } sectioning than I'm prepared to spend. } } Any ideas would be most welcome! } } Diana } } } } Diana van Driel } Dept Ophthalmology } Sydney University } GPO Box 4337 } Sydney, NSW } AUSTRALIA 2001 } } Phone 61 2 93827278 } Mobile 0423 151614 } FAX 61 2 93827318 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 2 08:27:30 2005
Diana, Try putting the ideas from Paul Webster and Judy Murphy together...if you've got mostly basement membrane, then a cationic dye, like alcian blue or ruthenium red will take quite nicely. You may need to play a bit with concentrations...both can get a little murky if you use too much. Years ago, I did some studies that looked at the proteoglycans in the heart using a variety of cationic dyes, as I recall we used concentrations that were on the order of 0.05-0.1% dye (wt-vol). Good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 2 09:07:23 2005
Hi, I wanted to know if anyone has much success with the hot glue gun sold by Cedarlane Labs. They cite a paper in the Proceedings of the 50th Annual Meeting of the EMSA/MAS/MSC.SMC in 1992, pages 410-411, to describe its merits for fast fixing of samples for SEM.
Thanks, Pat Research Technician SEM-FIB Facility Institute for Research in Materials Dalhousie University (902) 494-1258
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 2 11:32:09 2005
In my original message I gave a brief overview of how I handle small specimens. It is obvious that I missed out many details (as pointed out off-line) so here is a slightly more detailed description of how I do it.
First, the cell fragments/specimen/cells/junk etc is fixed in aldehyde. It is then pelleted down and washed with buffer containing glycine. The glycine reacts with the aldehyde and stops it cross-linking the gelatin.
I then mix the pellet with a small amount of 10% gelatin (for those in the US, Knox gelatin works really well, for the rest of the world, try food quality gelatin it doesn't precipitate phosphates). The gelatin is warm (37 degrees) and should remain warm while the specimen is mixed with it and centrifuged down. The pellet can be centrifuged into a small tube with a pointed base, or placed using a fixed angle rotor. Cooling the gelatin will make it gel (I know it is obvious but apparently it needed to be stated).
Once the gelatin has set, the pellet is cut out and fixed again in aldehyde. It can then be treated as a piece of tissue and processed in any way you like. If it is to be embedded in epoxy resin, then osmium treatment it recommended. This will turn the gelatin brown and make it easy to see in the polymerized plastic.
If Lowicryl or LR White embedding is required, then I would suggest using agarose embedding as an alternative. However, gelatin is liquid at 37 degrees but agarose has to be heated to 60 degrees, which may affect antigenicity. To make the block visible in the resin I add color to the agarose or gelatin. I do try to avoid staining the specimen.
An alternative approach to handling really small specimens is to mix them with something that bulks up the pellet but is easily identified in the pellet. RBC membranes make a good padding for nucleated cells and there is no way you can mix one up for another.
You could also mix your small sample with fibrin and polymerize it using fibrinogen (see Acta Histochem. 1998 Jul;100(3):309-13. Processing of free cells for electron microscopy using a fibrin clot. Raska I, Pliss A, Mandys V, Risueno MC, Lojda Z. for details of this). The clot is easy to handle and bulks up the pellet sufficiently to see it when it is embedded.
Finally, if you pellet the specimen, embed it in gelatin as I described above, you can take out the pellet and place it onto a glass slide covered with Parafilm. Pour warm gelatin over the pellet and press a second Parafilm-coated slide over the first. Use spaces to create a space between the two slides so that when you take the slides apart after the gelatin has gelled, you will have your specimens embedded in a thin film of gelatin. You may be surprised by how easily the specimen can be seen in this thin film. Cutting out the specimen is very easy, but remember to keep the gelatin cool or it will become liquid.
Good luck,
Paul Webster.
On 2/2/05 6:24 AM, "Leona Cohen-Gould" {lcgould-at-med.cornell.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Diana, } Try putting the ideas from Paul Webster and Judy Murphy together...if } you've got mostly basement membrane, then a cationic dye, like alcian } blue or ruthenium red will take quite nicely. You may need to play a } bit with concentrations...both can get a little murky if you use too } much. Years ago, I did some studies that looked at the proteoglycans } in the heart using a variety of cationic dyes, as I recall we used } concentrations that were on the order of 0.05-0.1% dye (wt-vol). } Good luck, } Lee
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 2 14:48:20 2005
Sigma makes low-temperature gelling agarose. Type VII has a gel point of about 30oC. We use this routinely for all samples...even those for ICC. We just dissolve the agarose (usually a 1.5% solution) and then cool it to about 40oC before adding it to samples. This is well below the temperature that should affect antigenicity but is still high enough to keep the agarose fluid while you are pelleting. I try to put the tubes in warm water when possible(depending on centrifuge) while spinning to also slow down the gelling.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 2/2/05 12:28 PM, "Paul Webster" {pwebster-at-hei.org} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hello again, } } In my original message I gave a brief overview of how I handle small } specimens. It is obvious that I missed out many details (as pointed out } off-line) so here is a slightly more detailed description of how I do it. } } First, the cell fragments/specimen/cells/junk etc is fixed in aldehyde. It } is then pelleted down and washed with buffer containing glycine. The glycine } reacts with the aldehyde and stops it cross-linking the gelatin. } } I then mix the pellet with a small amount of 10% gelatin (for those in the } US, Knox gelatin works really well, for the rest of the world, try food } quality gelatin it doesn't precipitate phosphates). The gelatin is warm (37 } degrees) and should remain warm while the specimen is mixed with it and } centrifuged down. The pellet can be centrifuged into a small tube with a } pointed base, or placed using a fixed angle rotor. Cooling the gelatin will } make it gel (I know it is obvious but apparently it needed to be stated). } } Once the gelatin has set, the pellet is cut out and fixed again in aldehyde. } It can then be treated as a piece of tissue and processed in any way you } like. If it is to be embedded in epoxy resin, then osmium treatment it } recommended. This will turn the gelatin brown and make it easy to see in the } polymerized plastic. } } If Lowicryl or LR White embedding is required, then I would suggest using } agarose embedding as an alternative. However, gelatin is liquid at 37 } degrees but agarose has to be heated to 60 degrees, which may affect } antigenicity. To make the block visible in the resin I add color to the } agarose or gelatin. I do try to avoid staining the specimen. } } An alternative approach to handling really small specimens is to mix them } with something that bulks up the pellet but is easily identified in the } pellet. RBC membranes make a good padding for nucleated cells and there is } no way you can mix one up for another. } } You could also mix your small sample with fibrin and polymerize it using } fibrinogen (see Acta Histochem. 1998 Jul;100(3):309-13. Processing of free } cells for electron microscopy using a fibrin clot. Raska I, Pliss A, Mandys } V, Risueno MC, Lojda Z. for details of this). The clot is easy to handle and } bulks up the pellet sufficiently to see it when it is embedded. } } Finally, if you pellet the specimen, embed it in gelatin as I described } above, you can take out the pellet and place it onto a glass slide covered } with Parafilm. Pour warm gelatin over the pellet and press a second } Parafilm-coated slide over the first. Use spaces to create a space between } the two slides so that when you take the slides apart after the gelatin has } gelled, you will have your specimens embedded in a thin film of gelatin. You } may be surprised by how easily the specimen can be seen in this thin film. } Cutting out the specimen is very easy, but remember to keep the gelatin cool } or it will become liquid. } } Good luck, } } Paul Webster. } } } } On 2/2/05 6:24 AM, "Leona Cohen-Gould" {lcgould-at-med.cornell.edu} wrote: } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------} } } - } } } } Diana, } } Try putting the ideas from Paul Webster and Judy Murphy together...if } } you've got mostly basement membrane, then a cationic dye, like alcian } } blue or ruthenium red will take quite nicely. You may need to play a } } bit with concentrations...both can get a little murky if you use too } } much. Years ago, I did some studies that looked at the proteoglycans } } in the heart using a variety of cationic dyes, as I recall we used } } concentrations that were on the order of 0.05-0.1% dye (wt-vol). } } Good luck, } } Lee } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 2 15:57:10 2005
The new Nanotech America site just launched this morning at www.nanotech-america.com. If you look under "How SPMs work", you will find a animations of all the major imaging and measurement modalities. Also, there is a textbook which can be downloaded under The Library.
Hope this is helpful.
Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 08:40 PM 2/1/2005, Palash.Gangopadhyay-at-fys.kuleuven.ac.be wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Feb 2 18:07:45 2005
Dear Patricia, I don't know about the hot glue gun sold by Cedarlane, but I have been using a household-type hot glue gun to fix samples on an SEM stub for twenty years. (I've been through several guns and learned to get the more expensive ones.) I find it is good for heavier or odd-shaped samples and gives a more secure hold with no creep than the sticky tabs, but it is not conductive, so you have to make a path to ground over the glue after it is hard. I use a stripe of carbon paint. What is your problem with it? Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Patricia Scallion" {PSCALLIO-at-DAL.CA} To: {Microscopy-at-MSA.Microscopy.com} Sent: Wednesday, February 02, 2005 7:05 AM
(Sorry about all of the } } } , please ignore them. ) } } } Diana, } } } This sounds like a job for the Formvar sandwich. Here is a } } } description I wrote as applied to the tiny arabidopsis root tip, but } } } the problem is the same. If you have further questions, please email. } } } Note that Formvar stands up just fine to acetone and epoxies but } } } propylene oxide will dissolve it. Good luck. TB. } } } } } } } } } The small size of arabidopsis roots (ca 0.15 mm diameter) makes them } } } easy to lose while changing solutions. To retain the roots, I use a } } } method that is not only convenient but also turns out to be } } } beneficial for sample preservation. Originally, I encased each root } } } in a small droplet of low-gelling-temperature agarose, but this is } } } messy and exposes the sample to heat, albeit briefly. Then, I } } } modified a method from cryofixation where samples are mounted on a } } } Formvar film. A chemically fixed root tip is placed on a } } } Formvar-coated wire loop, a second Formvar film secures the root tip } } } on the loop. The Formvar films are readily permeated by solvents and } } } small molecules. Between Formvar films, the thin arabidopsis root tip } } } is prevented from bending or twisting. I call this "mechanical } } } fixation" and beyond being convenient, it seems to enhance sample } } } preservation. } } } Loops are made in advance and coated by casting Formvar } } } rectangles (measuring a little more than the loop diameter on one } } } side and a little more than twice the loop diameter on the other) and } } } plunging the loop into the water over the rectangle so that the plane } } } of the loop bisects the long axis of the rectangle. The Formvar } } } rectangle wraps around the wire loop and the coated loop is removed } } } at once from the water. Such loops remain stable for months. To } } } secure a sample, the procedure is repeated: After the sample has been } } } fixed and rinsed, a loop (already Formvar coated) is placed } } } horizontally on a drop of water (or buffer) and the sample placed on } } } the Formvar. Excess sample is trimmed if needed, and the loop (with } } } sample) is plunged onto a new Formvar rectangle, thus encasing the } } } sample between Formvar layers, held by the loop. Several loops can be } } } placed in a vial and solutions exchanged without losing the sample. } } } The loop is embedded with the sample, and removed during trimming. I } } } use fine copper wire (36 gauge), which can be trimmed along with the } } } block. } } } } } } } } } } } } } } Hi all, } } } } } } } } I've been given some vanishingly small specimens to embed for TEM. } } } } They're peels from the surface of a retina - at most several cells } } } } thick and usually less than 1x2 mm in area. They are also made } } } } mostly of basement membrane, with a few whole cells and some cell } } } } fragments, so not only are they almost invisible to start with, but } } } } they remain invisible even after osmium. Pinning them down won't } } } } work - they're too small. I've embedded some once using a dissecting } } } } microscope, but it was a lot of work and even so I lost 2 of the 6. } } } } I thought of embedding first in agar, but then I'll end up with an } } } } invisible specimen somewhere in the resin, which will take longer } } } } find by serial sectioning than I'm prepared to spend. } } } } } } } } Any ideas would be most welcome! } } } } } } } } Diana } } } } } } } } } } } } } } } } Diana van Driel } } } } Dept Ophthalmology } } } } Sydney University } } } } GPO Box 4337 } } } } Sydney, NSW } } } } AUSTRALIA 2001 } } } } } } } } Phone 61 2 93827278 } } } } Mobile 0423 151614 } } } } FAX 61 2 93827318 _ ____ __ ____ / \ / / \ / \ \ Tobias I. Baskin / / / / \ \ \ Biology Department /_ / __ /__ \ \ \__ 611 N. Pleasant St. / / / \ \ \ University of Massachusetts / / / \ \ \ Amherst, MA, 01003 / / ___ / \ \__/ \ ____ Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243 http://www.bio.umass.edu/biology/baskin/ --
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 00:50:33 2005
Our campus-wide, multi-user microscopy imaging facility is considering the purchase of an instrument that would process slides for in situ hybridization or antibody staining (e.g., Intavis InsituPro VS). I am skeptical on the usefulness of these type of instruments in a multi-user environment where the reagents and staining conditions change on a daily basis. Are there any core facilities out there that are successfully using one? Do you recover the costs of consumables and make enough to cover the service contract and staff time? Your public or private responses would be welcome. Thanks, Tom Phillips
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Dear List, I am trying to find the best value for the lattice constant of gold at 82 K. I have only found the values for room temp and above. Does anyone have a reference for this value at low temperature? TIA. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 12:31:25 2005
Dear Colleagues: I am considering the Orion 6 system for passive digital image collection from my old analog SEM. Does anyone have personal experience (positive or negative) with the Orion system they would care to share? I would be happy to have your candid comments off-list. Thank you. --Jan Factor
---------------------------------------2/3/05 Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 13:28:49 2005
Dear Listers Thank you for you many(!) helpful and useful responses. Our researcher is overwhelmed by your generous help.
Richard Harris Laboratory Supervisor Department of Biology University of Western Ontario London Ontario CANADA N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 13:53:02 2005
I would be most hesitant to get one of these instruments and put it in a multi-user environment. We have the InSituPro unit (previous generation) for whole mount staining. We do all of the staining with this and charge back what it costs to do the service. We do not charge for staff time. One of the PIs here also has a unit, and they have one dedicated operator in their lab. The post-docs will bring their samples and probes, she will load them and do the runs, and then give them back to the post-doc for the chromogen step.
I caution against getting the VS for staining on slides as this application is still undergoing development. Personally I would prefer to wait until it has finished development and undergone field testing for a bit before committing $$$s to the instrumentation. I think the technology has promise and was very much encouraged by what I saw at the demo.
In my opinion, the best results from equipment such as this require dedicated operators. Yes, many people can be trained on using the instrument because it's not rocket science. But too many cooks that have their hands in the pot (so to speak), is a recipe for disaster. One day someone will come in and someone previously has toyed with the settings just enough that their run will be ruined. Or you'll have all your samples ready to run, only to find out the machine is not working properly, won't initialize, whatever, and have no indication from the previous user there was a problem.
But then again, maybe I'm just a control freak. LOL
Best of luck to you,
Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj-at-stowers-institute.org
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Thursday, February 03, 2005 11:02 AM To: Microscopy-at-msa.microscopy.com
Our campus-wide, multi-user microscopy imaging facility is considering the purchase of an instrument that would process slides for in situ hybridization or antibody staining (e.g., Intavis InsituPro VS). I am skeptical on the usefulness of these type of instruments in a multi-user
environment where the reagents and staining conditions change on a daily
basis. Are there any core facilities out there that are successfully using one? Do you recover the costs of consumables and make enough to cover the service contract and staff time? Your public or private responses would be welcome. Thanks, Tom Phillips
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Apologies in advance for double post to confocal and microscopy list.
The group studying flour and dough structure here wants to use FITC to stain the dough protein so they can get an idea of the macrostructure of the dough. The protocol has been published in a couple of papers from another group, and after staining the blob of dough they looked at it under confocal to see changes in fluorescence pattern after various amounts of mixing, and assumed that any fluorescence came from protein aggregates. This worries me, as I wasn't aware that FITC per se was such a specific stain for protein, and the published protocol says nothing about washing out excess free dye. The alternative I thought of was to use fluorescamine, which is only fluorescent when bound to protein, so you could put your sample into the dye and observe without rinsing. (Needs UV, though.)
Does anyone have experience with this sort of material, comments about FITC as a protein stain?
TIA, Rosemray
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 16:13:10 2005
Is the Intavis InsituPro VS similar to the automated immunohistochemistry and In Situ units sold by Ventana Medical Systems? The Ventana systems were developed at the University of Arizona and they seem to work well if you are doing the same sort of staining runs on histologic samples (paraffin or frozen sections) over and over again. I gather that there's a bit of a learning curve to know how to program in a new procedure.
Doug
Caveat: I have no financial interest in VMS, but I do know the founders and have several friends who work for the company. .................................................................... Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy Research Specialist, Principal University of Arizona (office: AHSC 4212A) P.O. Box 245044 (voice: 520-626-2824) Tucson, AZ 85724-5044 USA (FAX: 520-626-2097) (email: Cromey-at-Arizona.edu) .................................................................... http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 17:07:56 2005
Dr. Phillips, I have 30 years of experience in histology and close to 6 years of experience doing antibody staining. I don't know what the environment of your lab is, but in any lab where someone is doing a LOT of IHC (antibody staining) or ish (in situ hybridization) staining, automation is the way to go. The technology of these stainers has become really good and many of the bugs in the older instruments that you may be thinking of have pretty much been worked out. I'm not familiar with the stainer you mentioned, but I am familiar with Ventana Medical Systems and DAKOCytomation. IMO, DAKO makes one of the best and also happens to be the least costly because you can use whatever reagents you choose. Ventana makes a fabulous machine, but the one I used in my former job was tied in to the use of their reagents only. Their detection kits run over 2000USD per kit and each kit performs only 250 tests. I think Ventana now makes another stainer that is more reagent flexible. THey're a great company, just really expensive. I cannot advise you on whether it is economical for your lab to invest in this as I don't know how much work you're doing and what you charge per slide vs your overhead in salaries, etc. You can figure that out easily enough.
If there are to be mutliple users of these instruments, that is usually no problem as the manufacterer usually provides a training session for all employees. When you get your stainer, I would mandate that all techs who will use the instrument must be trained first. In most facilities, training is done not only when a new instrument arrives, but also a periodic check with each tech to ensure they are doing things right.
I hope this is helpful
Connie McManus Mt Ogden Scientific Services 950 W Kershaw, Suite E Ogden UT 84401 tel: 866/933-6677 conniemoss-at-relia.net www.mtogdensci.com (not yet available)
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 20:28:41 2005
Thanks to everyone. I now have a collection of ideas to try! Could the person who suggested blue material and who had the PowerPoint presentation please resend me the Email and I would love the presentation as well; somehow I managed to delete your message.
Diana
Diana van Driel Dept Ophthalmology Sydney University GPO Box 4337 Sydney, NSW AUSTRALIA 2001
I'm a graduate student from India. I'm working on digital holographic imaging. I am intereted to extend it for microscopic imaging . Recently I came to know that, there are simulated specimens (both amplitude and phase objects- phantoms-synthetic) available for microscopy experiments. Are there any suppliers where I can place orders for such objects according to my requirements or may be for the readily available samples.
Hoping your replies Sincerely, Anith.N
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From MicroscopyL-request-at-ns.microscopy.com Thu Feb 3 23:56:27 2005
Last year I posed a question on whether anyone had a method or software that could read and batch export ISIS maps and images from a job rather than doing it individually. I got great support from several individuals and now have a some great pieces of software that I use regularly. As a follow on I now need to do the same with the ISIS spectra. Is there anyone out there who has a method or software for reading the ISIS spectra and batch saving the files as txt, tif or jpg.
Any help would be greatly appreciated.
Regards George
George Theodossiou Physicist / Microscopist Microscopy and Microanalysis Laboratory
AMCOR Research and Technology Ph: +61 3 9490 6135 Fax: +61 3 9499 4295 Mobile: 0409 568 840 email: George.Theodossiou-at-amcor.com.au
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From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 07:23:13 2005
I come from an EM background and have recently assumed responsibility for LM evaluations. However, I am in need of a reference detailing what stains what, and for which types of microscopy. Our current microscope is equipped for UV/fluorescence (among other things), and this is of primary interest to me at this point.
So if anyone can point me toward an LM staining reference, I would greatly appreciate it. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
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From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 08:26:06 2005
Though not an on-line course, you should consider the AFM short course (one week) at NC State University. Check it out at http://www.ncsu.edu/aif/afmcourse
Lehigh U also has a course but they cancelled it last year so I am not sure if it will be held this year or not.
On Tue, 1 Feb 2005 20:40:39 -0600 Palash.Gangopadhyay-at-fys.kuleuven.ac.be (by way of Ask-A-Microscopist) writes: } } } ------------------------------------------------------------------------- ----- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------- ------ } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (Palash.Gangopadhyay-at-fys.kuleuven.ac.be) from } http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Tuesday, February 1, 2005 at 12:27:56 } ------------------------------------------------------------------------- -- } } Email: Palash.Gangopadhyay-at-fys.kuleuven.ac.be } Name: Dr. Palash Gangopadhyay } } Organization: Molecular and Nano Materials, K U Leuven } } Education: Graduate College } } Location: Leuven, Belgium } } Question: Dear Microscopists } } I will appreciate very much if somebody can direct me to an online } AFM / MFM / STM course immediately available. } } Thanks in Advance } Palash Gangopadhyay } } ------------------------------------------------------------------------- -- } } }
Richard Fiore, Southeast Sales Carl Zeiss SMT (formerly LEO Electron Microscopy) 5127 Orange Grove Rd., Hillsborough, NC 27278 Tel: 919-643-2234, Cell: 919-593-1960, Fax: 919-643-2667 Email: rfiore-at-juno.com or fiore-at-smt.zeiss.com, URL: www.smt.zeiss.com/nts
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 08:43:34 2005
Ditto what Connie said about the Dako instrument...it seems to be the "industry standard" for automated IHC. However, it does not have capability for doing in situ hybridization. We currently have the Ventana Discovery to do the automated ISH on slides, in addition to the InSituPro for doing automated ISH on whole mounts. What makes the new InSituPro VS unit so compelling is that you can do either whole mounts or slide-mounted tissues on it on any given run.
To further clarify my position on multiple users, in our lab all technicians are trained on every instrument. My initial response to Dr. Phillips about having dedicated users was with regard to not making it available for post-docs and other researchers to just come in and run the thing, which is a common practice in core facilities.
Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj-at-stowers-institute.org
-----Original Message----- } From: Connie McManus [mailto:conniemoss-at-relia.net] Sent: Thursday, February 03, 2005 5:06 PM To: microscopy-at-msa.microscopy.com
Dr. Phillips, I have 30 years of experience in histology and close to 6 years of experience doing antibody staining. I don't know what the environment of your lab is, but in any lab where someone is doing a LOT of IHC (antibody staining) or ish (in situ hybridization) staining, automation is the way to go. The technology of these stainers has become really good and many of the bugs in the older instruments that you may be thinking of have pretty much been worked out. I'm not familiar with the stainer you mentioned, but I am familiar with Ventana Medical Systems and DAKOCytomation. IMO, DAKO makes one of the best and also happens to be the least costly because you can use whatever reagents you choose. Ventana makes a fabulous machine, but the one I used in my former job was tied in to the use of their reagents only. Their detection kits run over 2000USD per kit and each kit performs only 250 tests. I think Ventana now makes another stainer that is more reagent flexible. THey're a great company, just really expensive. I cannot advise you on whether it is economical for your lab to invest in this as I don't know how much work you're doing and what you charge per slide vs your overhead in salaries, etc. You can figure that out easily enough.
If there are to be mutliple users of these instruments, that is usually no problem as the manufacterer usually provides a training session for all employees. When you get your stainer, I would mandate that all techs who will use the instrument must be trained first. In most facilities, training is done not only when a new instrument arrives, but also a periodic check with each tech to ensure they are doing things right.
I hope this is helpful
Connie McManus Mt Ogden Scientific Services 950 W Kershaw, Suite E Ogden UT 84401 tel: 866/933-6677 conniemoss-at-relia.net www.mtogdensci.com (not yet available)
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 10:57:23 2005
If fluorescence is your primary interest I would strongly recommend that you visit websites for the following companies: Molecular Probes - they make the broadest range of fluorescence probes and have very rich content as well as gorgeoous images Quantum Dots - a new approach to fluorescent staining. Instead of being based on organic molecules, the qDots are derived from more inert, semiconductor based materials.
The two primary filter manufacturers in our industry are Omega Filters (www.omegafilters.com) and Chroma. Both have, again, very informative websites.
Re: learning more about fluorescence - I would encourage you to visit the Nikon mini-university, Olympus' site, and (at the risk of being a bit self-serving), getting a copy of Optimizing Light Microscopy (visit www.microscopyeducation for details).
Hope this was helpful Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
Caveat: MME has no financial involvement in any of the companies mentioned above.
At 07:21 AM 2/4/2005, Mr Brian Kirkmeyer wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 11:42:56 2005
I assume you want to stain something with a fluorescent antibody of some kind. This is not in my area of expertise, but from what I understand, before you do that, you need to have some kind of stained section to get a fix on where you are in the tissue. Also, it is important to know what kind of tissue prep ... FFPE (formalin fixed paraffin embedded) or are the tissues processed in plastic? Are these sectioned at 3-5 um? more or less that that? Do you want special stains ((i.e. stains for connective tissues, glycogen, fungi, bacteria, mast cells ... ad infinitum) or do you just want a plain, simple stain that will show the basic cell arrangements in the tissue ???? What do you want to see? There are some excellent texts out there. Perhaps the best one for starters is Frieda Carson's book on Histotechnology. Can be purchased from Sigma, Amazon.com, Barns Noble, etc. and it's fairly inexpensive. There are tons and tons of really good histology books out there. Also, it doesn't hurt to have a sit down chat with your technicians and find out from them what they can do and what they would recommend.
I'm sorry not to be of more help. Please feel free to contact me privately if you need help of any kind.
Connie McManus Mt Ogden Scientific Services 950 W Kershaw, Suite E tel: 866/933-6677 fax: 435/514-1781 conniemoss-at-relia.net www.mtogdensci.com
------------------------------------------ } } Kirk wrote: { { I come from an EM background and have recently assumed responsibility for LM evaluations. However, I am in need of a reference detailing what stains what, and for which types of microscopy. Our current microscope is equipped for UV/fluorescence (among other things), and this is of primary interest to me at this point.
So if anyone can point me toward an LM staining reference, I would greatly appreciate it. Thanks!
Kirk
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 16:35:09 2005
George Theodossiou {George.Theodossiou-at-amcor.com.au} wrote: } } ...Is there anyone out there who has a method or software for reading } the ISIS spectra and batch saving the files as txt, tif or jpg. } I have written some software that will convert ISIS spectra to JPEG, BMP or text format. You can convert files singly or as a batch. The programme also reads, displays and converts other Link/Oxford formats, as well as Noran, Edax and EMSA formats.If you can give me your mailing address I will send you a copy.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 17:54:10 2005
Several of you have written re: problems geting to www.microscopyeducation.com. Much to my chagrin, the links mentioned in my earlier posting got crossed with those of our sister group. I've spoken to the webmaster and he will have it fixed over the weekend. Please try again. ... Follow the Nav buttons in The Library to link to Optimizing Light Microscopy.
Thanks :-) Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 10:54 AM 2/4/2005, Barbara Foster wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 4 19:20:51 2005
Is there a separate formal list for Zeiss/LEO Supra users? If so, I'd appreciate knowing about it. If not, I would like to communicate with others about discoveries, disappointments, observations, history, experiences, lessons learned (easy and hard way) and other topics relative thereto.
We can keep this off of the MSA list and I would forward all relevant material to those who are interested.
This is for Supra and SupraVP.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 5 06:29:34 2005
Dear friends, I am looking for infos for setting up a FIB, dual scan, system. All my best AD
------------------------------------------------------------------------ ------------------------------------------------------------------------ -------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001)
------------------------------------------------------------------------ ------------------------------------------------------------------------ ----------------------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html
------------------------------------------------------------------------ ------------------------------------------------------------------------ ----------------------------------------------------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001)
------------------------------------------------------------------------ ------------------------------------------------------------------------ ----------------------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html
What information exactly are you looking for? If you have specific questions I may be able to help you out.
Disclaimer: PBS&T provides support for charged particle beam instruments commercially.
Cheers, ================= Valery Ray www.partbeamsystech.com
--- Alberto Diaspro {diaspro-at-fisica.unige.it} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear friends, } I am looking for infos for setting up a FIB, dual } scan, system. } All my best } AD } } } } ------------------------------------------------------------------------ } } ------------------------------------------------------------------------ } } -------------- } [...] Dance with wolves and count the stars, } including the } unseen...(L.Ferlinghetti, Challenges to young poet, } 2001) } } ------------------------------------------------------------------------ } } ------------------------------------------------------------------------ } } ----------------------------------------------------------- } Alberto Diaspro, MicroScoBIO Research Center, } LAMBS-IFOM, Department } of Physics, University of Genoa, Via Dodecaneso 33, } 16146 Genova, Italy } facsimile +39-010314218 - voice } +39-0103536426/480/309; URL: } http://www.lambs.it } } http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html } } ------------------------------------------------------------------------ } } ------------------------------------------------------------------------ } } ------------------------------------------------------------ } } } } ------------------------------------------------------------------------ } } ------------------------------------------------------------------------ } } ----------------------------------------------------------- } [...] Dance with wolves and count the stars, } including the } unseen...(L.Ferlinghetti, Challenges to young poet, } 2001) } } ------------------------------------------------------------------------ } } ------------------------------------------------------------------------ } } ----------------------------------------------------------- } Alberto Diaspro, MicroScoBIO Research Center, } LAMBS-IFOM, Department } of Physics, University of Genoa, Via Dodecaneso 33, } 16146 Genova, Italy } facsimile +39-010314218 - voice } +39-0103536426/480/309; URL: } http://www.lambs.it } } http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html } } ------------------------------------------------------------------------ } } ------------------------------------------------------------------------ } } ------------------------------------------------------------ } } }
===== Valery Ray
Particle Beam Systems & Technology www.partbeamsystech.com
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 5 11:25:30 2005
As was pointed out to me, the Gemini column is also on the LEO 1550. So these would apply as well. I suppose the only main differentiating factor is plinth and uni-plinth. But the column seems to be the same.
gary g.
At 05:19 PM 2/4/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Feb 5 19:16:24 2005
Has anyone done any measurements on the resistivity of Os coatings? Sheet rho vs. thickness, etc.?
If a charging specimen was imaged at 20-25KV, what Os coating thickness would eliminate the charge? Then, given that thickness, what would the inherent sheet resistance be?
gary g.
From MicroscopyL-request-at-ns.microscopy.com Sun Feb 6 08:45:14 2005
To all of you who informed me about your experiences, tips & tricks etc. concerning calibration of objectives, thanks a lot for the massive input! This states again that this listserver is not just 'a bunch of people', but a nice collection of pro's with some pro's-to-become, just as myself (hopefully), who can learn a lot from them!
I calibrated every objective individually with a stage micrometer, but still noticed about + or - 4% deviation between the 2,5x and 40x objective. Thanks to your input I now know that it is not so bad, and, I recalibrated the objectives, but more accurate, and now have about 2,1% difference. Not too bad I think, though it took some time!
Thanks again for your answers, you were of great help!
Sven Terclavers
"A microscopist observes things which nature hides for the majority, always feel honoured for that!"
From MicroscopyL-request-at-ns.microscopy.com Sun Feb 6 14:52:40 2005
Alberto, What is it that you are looking for exactly? Is it for setting up different scans etc for an existing FIB ? or something else. You can reply on line or offline, Best regards, Bobby Hooghan Hooghan-at-grandecom.net
-----Original Message----- } From: Alberto Diaspro [mailto:diaspro-at-fisica.unige.it] Sent: Saturday, February 05, 2005 6:28 AM To: MSA listserver
Dear friends, I am looking for infos for setting up a FIB, dual scan, system. All my best AD
------------------------------------------------------------------------ ------------------------------------------------------------------------ -------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001)
------------------------------------------------------------------------ ------------------------------------------------------------------------ ----------------------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html
------------------------------------------------------------------------ ------------------------------------------------------------------------ ----------------------------------------------------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001)
------------------------------------------------------------------------ ------------------------------------------------------------------------ ----------------------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html
There seems to be some interest in a focused group for Zeiss/LEO Gemini systems. I have formed a user group on Yahoo that is open to all Gemini column system users (if you don't have a Gemini column system, you likely won't be interested in this topic/group).
The Gemini systems include all LEO 1500 and Supra models which are high vacuum and/or variable pressure (VP). The purpose of the group is to exchange experiences and info on these systems. Contact info, service experience, system nuances, macro editing and creation, etc. are key topics. The group is not moderated. I started the group but I do not moderate or edit it. All that is required is vetting-- you need to apply for group membership and then you are in.
Group Settings: -listed in directory (does not work at this time--no idea why not) -membership requires approval -messages do not require approval -all members may post messages -message archive viewable by members only -e-mail attachments are not permitted
Hopefully this is useful for Gemini users. As a newbee, I hope to gain from others' experiences. Perhaps, others can gain from my experiences. A goal is to not clutter up the MSA list with system-specific threads. But of course, you can post this topic if you so desire. General EM topics still should go to the MSA list. The gemini list should be specific to this Gemini group of systems (no EVSEM, etc.).
} From the responses I have received so far, it seems that I should clarify what I'm looking for a little bit more. I seek a reference that describes staining/dyeing/tagging methods for non-histological samples, as I work with flavor and fragrance chemical compounds. So recipes for tissue stains or tags, unless related to a specific chemistry, unfortunately don't do me a lot of good. If anyone knows of such a non-histological reference, I would love to hear of it.
While on the subject, I am also interested in knowing what stains/dyes/tags are safe for contact with human skin. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Kirk, } } I assume you want to stain something with a } fluorescent antibody of some } kind. This is not in my area of expertise, but from } what I understand, } before you do that, you need to have some kind of } stained section to get a } fix on where you are in the tissue. Also, it is } important to know what } kind of tissue prep ... FFPE (formalin fixed } paraffin embedded) or are } the tissues processed in plastic? Are these } sectioned at 3-5 um? more or } less that that? Do you want special stains ((i.e. } stains for connective } tissues, glycogen, fungi, bacteria, mast cells ... } ad infinitum) or do you } just want a plain, simple stain that will show the } basic cell arrangements } in the tissue ???? What do you want to see? There } are some excellent } texts out there. Perhaps the best one for starters } is Frieda Carson's } book on Histotechnology. Can be purchased from } Sigma, Amazon.com, Barns } Noble, etc. and it's fairly inexpensive. There are } tons and tons of } really good histology books out there. Also, it } doesn't hurt to have a } sit down chat with your technicians and find out } from them what they can } do and what they would recommend. } } I'm sorry not to be of more help. Please feel free } to contact me } privately if you need help of any kind. } } Connie McManus } Mt Ogden Scientific Services } 950 W Kershaw, Suite E } tel: 866/933-6677 } fax: 435/514-1781 } conniemoss-at-relia.net } www.mtogdensci.com } } } ------------------------------------------ } } } Kirk wrote: { { } I come from an EM background and have recently } assumed } responsibility for LM evaluations. However, I am in } need of a reference detailing what stains what, and } for which types of microscopy. Our current } microscope } is equipped for UV/fluorescence (among other } things), } and this is of primary interest to me at this point. } } So if anyone can point me toward an LM staining } reference, I would greatly appreciate it. Thanks! } } Kirk } } } }
__________________________________ Do you Yahoo!? Yahoo! Mail - You care about security. So do we. http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 7 09:01:57 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spradhan-at-siu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, February 6, 2005 at 21:39:34 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver:Hitachi S2460 N stage
Question: Hello list! I have been looking around, pretty much in vain, for a spare door and stage assembly for a Hitachi S2460N SEM for quite sometime now...We wanted to have it so that we could modify it to suit our purpose for conducting some experiments at our facility here...However, the situation looks very grim to me since i have almost exhausted all possible resources i have know of...could you suggest some more places i could look for it? Or better still, do you think it would be possible to modify such assemblies of other hitachi models to fit our machine? i shall look forward to your suggestions and advice..thanks a lot in advance... regards, sailesh.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cvierret-at-umr.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, February 7, 2005 at 08:43:52 ---------------------------------------------------------------------------
I am looking for information on sample preparation of a Ge single crystal for TEM examination. I need to know how to cut the sample, thin the sample and any other information that would make this job simpler or at least less frustrating.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (medunn-at-mail.uri.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, February 7, 2005 at 08:39:20 ---------------------------------------------------------------------------
Email: medunn-at-mail.uri.edu Name: Michael Dunn
Organization: Uinversity of Rhode Island
Education: Graduate College
Location: Kingston, RI
Question: 1)Is there a way to quanitfy the number of cells contained in a tissue sample that is being analyzed on the electron microscope through the use of a software program like NIH?
2)What is the correlation, if any, with the mitochondrial density of a section and the cell count of that same section?
Tom: Please re-send the email you sent to me over the weekend. (Between home and work, it ended up on the wrong computer. - Sorry to use the list for this.) --Thanks, Jan
---------------------------------------2/7/05 Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College, State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 7 10:57:01 2005
I'd suggest that you contact Chuck Garber at SPI (www.2SPI.com). He is a wealth of info in this area.
Good hunting! Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
Caveat: MME has no financial interest in this product.
At 07:14 PM 2/5/2005, Gary Gaugler wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Feb 7 11:03:35 2005
Again, I would really recommend that you investigate fluorescence. The chemistry is very specific.
As for safety, again, the probe companies have all of that information.
Good hunting, B
At 07:41 AM 2/7/2005, Mr Brian Kirkmeyer wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Feb 7 14:45:37 2005
I am not speaking from direct experience for doing Ge, but offhand, Ge should not be more difficult than silicon. If it is a single crystal of Ge, you should know the crystallographic directions of the sample. Of the following, the two least expensive and easy methods would be chemical polishing and small angle cleavage technique.
I would look at several standard techniques to start.
1) if you are doing plan view samples, chemical polishing should work nicely. I don't know the chemical polish for Ge, but I am sure that it is in the literature. Peter Goodhew has a simple chemical polisher apparatus in his book, Thin Foil Preparation for Electron Microscopy (Practical Methods in Electron Microscopy, Vol 11). Basically, it is a tilted rotating cup with a platform in the center where the chemical drips from a burret until the material is perforated. South Bay Technology sells a chemical polisher that can be set up specifically for accurate and reproducible termination, especially if Bernie Kestel's instructions are followed.
2) mechanical polishing, dimpling, and ion milling should work nicely. See the equipment brochures from South Bay Technology, E. A. Fischione, Baltec, and Gatan
3) Ge cleaves. For cross sections and bulk crystal the small angle cleavage technique (microcleave technique) would work very well on this. This gives the best samples known to man (and women). I gave a little short course at UMR a few years ago on SACT. Talk to Lou Ross about it and see South Bay Technology's web site on microcleave for information on this.
4) Tripod Polishing -again see South Bay Technology web site.
5) FIB -Pay some money and have no trouble at all.
-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: by way of MicroscopyListserver [mailto:cvierret-at-umr.edu] Sent: Monday, February 07, 2005 10:01 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cvierret-at-umr.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, February 7, 2005 at 08:43:52 ---------------------------------------------------------------------------
I am looking for information on sample preparation of a Ge single crystal for TEM examination. I need to know how to cut the sample, thin the sample and any other information that would make this job simpler or at least less frustrating.
Gary, Just want to clarify something that others might be confused about.
The inherent property to the material that is deposited is its resistivity, not sheet resistance. Sheet resistance is measured usually with a four point probe and has units of ohms per square. It is dependent on the thickness of the material. If you have a square of any size, it will have the same resistance. (It is amazing to see a large 4 foot by 4 foot piece of glass with a conductive coating on it measure the same resistance as a one inch by one inch piece.) If you multiply the sheet resistance by the thickness, you get the resistivity with units ohm-cm.
This all assumes that the layer is uniform across the surface. For very light layers, that may not be the case. For charge drainage, it also assumes that the charge can come to the surface and be drained off. If the conductivity of the sample material is so low that this can't happen, then the relatively high voltage that you are using would put charge deep into the sample. There your charging affects would be voltage dependent. There is an article on that in the latest Microscopy Today issue, isn't there?
-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