Apologies for the late reply. I don't want to overdo this topic, but few peeps have addressed one of the original questions: is there much danger to getting abrasive sloshed in your eye?
While SiC/water splashes might not be much of a chemical irritant, aqueous suspensions of metal oxide particles are typically not pH-neutral, e.g., alumina suspensions--} acidic, silica suspensions--} basic. So, if you work with those types of polishing suspensions, an eye splash seems likely to be more painful than just the irritation associated with having gritty stuff in your eye.
I work in a lab where I wear safety glasses and frequently go back and forth between the polisher and the optical microscope. It's a bit of a pain to take the glasses off and on, but you get used to it, and the slight inconvenience is worth preventing a splash in the eye.
Eve
-- Eve Donnelly Experimental Biomechanics Lab Sibley School of Mechanical & Aerospace Engineering Cornell University 130 Upson Hall Ithaca, NY 14853 tel. 607.255.3582 fax. 607.255.1222
I have two suggestions: 1. Cryo sections of the same should help 2. Application of alcohol helps to loosen the epithelium which can then be gently peeled off. This techique is used to debride the corneal epithelium during surgeries. Shashi CCMB, Hyderabad INDIA
Hi all,
I've been asked to see what the EM difference is between rat eye lens epithelium in culture and in situ. So I'd like to fix the epithelium while it is still in the eye. I'm worried that if I do that it then won't be possible to get the epithelium off the lens without damaging it; I also seem to remember that lens becomes brittle and difficult to handle when fixed. I'd like to keep life simple, so it may be necessary to take the epithelium off the lens before fixing. Does anyone have experience with this?
Thanks,
Diana
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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Temporary position (contractor status) offering 40 hrs/week with limited benefits
Job Description: Transmission Electron Microscopist / Analyst in Process Characterization Lab supporting the Advanced Technology Development Facility (ATDF) as well as both internal and external projects at International SEMATECH (ISMT) in Austin, Texas. The TEM group at ISMT provides materials characterization in support of research and process development projects for future integrated circuit manufacturing. Our research projects tend to focus on materials issues several years ahead of current semiconductor industry manufacturing trends, we therefore often study materials systems that offer new and exciting characterization problems and often collaborate with leading researchers at national labs and universities around the world, however we also have a significant workload involving routine characterization and process monitoring for systems that are already well understood.
Required: M.S. or Ph.D. in Materials Science, Solid-State Physics or Chemistry with strong analytical TEM experience.
For more details please contact: Brendan Foran -at- Brendan_Foran-at-SEMATECH.org
International SEMATECH, headquartered in Austin, Texas, is a global consortium of leading semiconductor manufacturers that represent about half the world's semiconductor production. International SEMATECH engages in cooperative, precompetitive efforts to improve semiconductor manufacturing technology through the support of our members. To learn more about International SEMATECH, visit our website at www.sematech.org.
} not related to the real safety (mostly keeping paperwork in a good shape } and accurately fill out chemical waste disposal tags). So, in such } situation, I think, it's superviser's personal responsibility to provide } safely environment to the workers and train them accordingly.
Look, we don't want to have this devolve into a session on what is safe and what isn't, but this is a very simplistic view. Paperwork may be essential for real safety of all handlers of material down the chain of disposal. If a supervisor has a limited budget, he/she is going to scrimp on safety.
Policies are in place to protect workers who historically have been mistreated. Yes, blanket policies do not apply in all situations. The bureaucracy simply needs some flexibility for individually documented exemptions.
} P.S. By the way: in my EM work, my glasses, I wear to correct my vision, } save my eyes in uncounted number of times. I, also, used to remove them } every time I am using light microscope or binoculars. I don't see big } problem removing them if necessary. But: yes, it decreases my } productivity. Ok, let say, I need about 5 sec. to remove them and put } back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day.
But you're wearing gloves and your gloves shouldn't be near your unshielded eyes each time you have to handle the googles. So you really should take off your gloves before removing your goggles each time.
____________________________________________________________________________ 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/
Dear Listeners, I am making ferrite nanoparticles doped with Co and/or Mn in the size range of 20-100nm. I have looked at them with TEM and made up size distributions from the pictures. However, I would now like to check my distribution curves with a particle sizer. Does anyone now if this is possible? I suspect it to be difficult since my particles are magnetic. If it is possible, does anyone know what type of medium I can disperse my particles in? So far I have tried to alter the pH but it doesn't seem to be enough and the particles still agglomerate before the the measurement is over. Thanks .
Yours sincerely Richard
PhD student, Swedish Defense Research Agency Department of Polymer Technology Royal Institute of Technology Teknikringen 56 SE-100 44 Stockholm SWEDEN phone: +46-8-790 76 37
--------------------------------------------------------- Legal Notice: This electronic mail and its attachments are intended solely for the person(s) to whom they are addressed and contain information which is confidential or otherwise protected from disclosure, except for the purpose they are intended to. Dissemination, distribution, or reproduction by anyone other than their intended recipients is prohibited and may be illegal. If you are not an intended recipient, please immediately inform the sender and send him/her back the present e-mail and its attachments and destroy any copies which may be in your possession.
Our webppage on connecting a Coolpix to a microscope will answer a lot of your questions: http://www.mvia.com/Coolpix/clpxadpt.htm
Also, please see my answers below...
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Alan Davis wrote: } } The Coolpix 5000 looks interesting. It does much of what I need, } including monitoring the live specimen on a TV, Computer, or video } projector. I like some of the features of the 5700, including better } telephoto range for shore bird studies. I have a couple of questions I } would refer to those with Coolpix experience: } } How well does the 5700 work on a scope?
Very poorly - you'll need to use the digital zoom mode on the 5700. See our FAQ section: http://www.mvia.com/Coolpix/clpxadpt.htm#FAQ
} Does the 5700 identically to the 5000 out of the box, with the Nikon } adaptor kit?
NO! You'll need to use the digital zoom mode on the 5700 as opposed to the optical zoom mode on the 5000, which will result in very poor images.
} Are these cameras significantly better than the 4500?
The 5000 has a higher resolution, but does not have a swivel body as the 4500 model does, which make it a little less ergonomical. However, the 500 will give you better images.
} Are macro kits available to enable focus to within 1/8"? (I intend } to use it as an "aquarium microscope" for example to study the community } at the sediment/water column interface, for which the Videoflex camera } works admirably well.)
There are a couple of lenses available from Nikon for various wide angle and fisheye applications.
} Can one take a shot easily an within ordinary lag values while } monitoring in real time?
Yes, but the live image will pause when you capture your image.
} Nikon has a relay lens (is that a projection lens?) for this } purpose, that will work on Coolpix 5000; does it work ok on the 5700? } } Another question: is the Nikon D-100 worth the extra cost, and does it } incorporate the video monitoring Coolpix features? Is it useable on a } scope, and how? } } Thank you for any comments or suggestions. I apologize for asking so } many questions at once... } } Alan Davis } Marianas High School } Saipan } } I wish to ask whether microscopists feel it } } -- } adavis-at-saipan.com 1-670-322-6580 } Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI } } I have steadily endeavored to keep my mind free, so as to give up any } hypothesis, however much beloved -- and I cannot resist forming one } on every subject -- as soon as facts are shown to be opposed to it. } -- Charles Darwin (1809-1882) } } The right to search for truth implies also a duty; one must not } conceal any part of what one has recognized to be true. } -- Albert Einstein } } As we enjoy great advantages from the inventions of others we should } be glad of an opportunity to serve others by any invention of } ours, and this we should do freely and generously. } -- Benjamin Franklin } }
Here is our answer to the many responses concerning the previous thread on Torr Seal. Torr Seal is an extremely high quality epoxy resin manufactured under a private label agreement as Torr Seal. Since there are non-critical applications which do not require such a high quality epoxy we, like others do carry a product known as a Torr Seal equivalent. We do let all our users know that when they select an 'equivalent they are paying less, but naturally there is a sacrifice in quality.
John Arnott
Disclaimer: Ladd Research sells the products mentioned in this e-mail.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Dear Listeners, I am making ferrite nanoparticles doped with Co and/or Mn in the size range of 20-100nm. I have looked at them with TEM and made up size distributions from the pictures. However, I would now like to check my distribution curves with a particle sizer. Does anyone now if this is possible? I suspect it to be difficult since my particles are magnetic. If it is possible, does anyone know what type of medium I can disperse my particles in? So far I have tried to alter the pH but it doesn't seem to be enough and the particles still agglomerate before the the measurement is over. Thanks .
Yours sincerely Richard
PhD student, Swedish Defense Research Agency Department of Polymer Technology Royal Institute of Technology Teknikringen 56 SE-100 44 Stockholm SWEDEN phone: +46-8-790 76 37
--------------------------------------------------------- Legal Notice: This electronic mail and its attachments are intended solely for the person(s) to whom they are addressed and contain information which is confidential or otherwise protected from disclosure, except for the purpose they are intended to. Dissemination, distribution, or reproduction by anyone other than their intended recipients is prohibited and may be illegal. If you are not an intended recipient, please immediately inform the sender and send him/her back the present e-mail and its attachments and destroy any copies which may be in your possession.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Jlindstrom-at-Hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, September 2, 2003 at 17:19:35 ---------------------------------------------------------------------------
Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
I would appreciate off-list feedback from users of JEOL 6700F tools.
I'm looking at a system with turbo pump, large specimen exchange interlock, solid state BSE vs. Robinson BSE, beam stability, accommodation to EDAX Genesis EDS and overall usability and support. Location is Northern CA.
Off-list please. All are confidential. Trying to make a purchase decision between this and comparable Hitachi. Specimens are microchips in cross section, top down, metallurgical coupons of IC manufacturing tools, legacy support for Amray 12mm diameter 3.1mm pin specimen stubs. Also, there is a need for working with rather large specimens. Like blocks of NIST standards (100mm diameter, 50mm high).
Native digital capture resolution is low. Are there other options that work? Is anyone using Soft Imaging ADDA? 4Kx4K pixels minimum is needed.
} From: "Michael Cammer" {cammer-at-aecom.yu.edu} : : } not related to the real safety (mostly keeping paperwork in a good shape : } and accurately fill out chemical waste disposal tags). So, in such : } situation, I think, it's superviser's personal responsibility to provide : } safely environment to the workers and train them accordingly. : : Look, we don't want to have this devolve into a session on what is safe and : what isn't, but this is a very simplistic view. Paperwork may be essential : for real safety of all handlers of material down the chain of disposal. If : a supervisor has a limited budget, he/she is going to scrimp on safety. : : Policies are in place to protect workers who historically have been : mistreated. Yes, blanket policies do not apply in all situations. The : bureaucracy simply needs some flexibility for individually documented : exemptions. : : } P.S. By the way: in my EM work, my glasses, I wear to correct my vision, : } save my eyes in uncounted number of times. I, also, used to remove them : } every time I am using light microscope or binoculars. I don't see big : } problem removing them if necessary. But: yes, it decreases my : } productivity. Ok, let say, I need about 5 sec. to remove them and put : } back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day. : : But you're wearing gloves and your gloves shouldn't be near your unshielded : eyes each time you have to handle the googles. So you really should take : off your gloves before removing your goggles each time. : Face shields will solve the glove problem. If the curve of plastic face shields interferes with your work a welders helmet with plain glass or you prescription lenses could be used. You would need some clearance behind the microscope but it would speed things up and not run the risk of getting something in your eye when you were removing goggles. With prescription lens in the helmet it should speed up work as you don't have to remove your glasses. It also solves the problem of scratched face shields.
Here is an example http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=2551210520&category=633 The part below the lens should be cut away as you don't need protection from the burning rays of the welders arc and the dark lens replaced with a clear one. Then just the push of your thumb will stand the helmet straight up out of the way of a microscope.
A welders helmet should be sturdy enough to satisfy any safety officer and quicker to tip up than removing goggles. Some surgery on the hood would make it lighter and reduce the clearance need behind the microscope with out compromising eye safety.
There are also some goggles that have a flip up lens available at welders supply houses as well. I don't know if they would clear a microscope. Here is an example http://cgi.ebay.com/ebaymotors/ws/eBayISAPI.dll?ViewItem&category=35000&item=2429373537 There are other designs that might be more suitable with the lens swinging 180 degrees. A call to your local Air Gas or other welding supply dealer will let you know what is available.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
Dear Colleagues, We have following trouble with Zephyr ZEM 1000CW watercooling unit: When our CM12 is switched on from STAND BY, overload protector switches off the pump motor of ZEM 1000CW from time to time. If the reset button of overload protector is pressed down, the motor starts and all is OK. But after several days or weeks this occurs again. Please, can you give us some hints or suggestions. Many thanks in advance. Oldrich Benada
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
Hello, I am searching for a manual for Reichert FC4 cryoultramicrotome. Is there anybody who could help me with that? Best regards,
Dr Dobroslawa Budka Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa fax: +27-21-8433543 phone: +27-21-8431161 mobile phone: 0722656091
We are looking for volunteers to perform beta testing of some code to do HREM image processing and Direct Methods. The code is a gnu-like package (i.e. free) which should compile on any unix or Mac computer fairly simply using a conventional "./configure ; make ; make install" strategy. If you know what I am talking about and are interested, please email me (not the whole list). Thanks.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm2-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I just about wrote "Urgent Assistance Needed" as the subject line, but then realized that would probably not make it past several dozen spam filters. I'm in my usual position (stuck between a rock and a deadline). I'm looking for a copy (original reprint or the journal issue) of:
MANTON I. & VON STOSCH H.A. 1966. Observations on the fine structure of the male gamete of the marine centric diatom Lithodesmium undulatum. Journal of the Royal Microscopical Society 85: 119-134.
I need to scan several figures from the paper, and our interlibrary loans can only get a xerox copy. Can anybody help? I'd gladly pay courier charges (both ways if you need the journal/paper back). Alternatively, if someone is willing, they could scan the figures from their end, but it would most likely be all of the plates in the paper, as I'm not sure right now which ones I need.
If you help, I'll make several offerings on your behalf to the gods of microscopy, requesting long filament life, long strings of buttery smooth silver-gray thin sections, and awesome powers against the Dark Forces of Bureaucracy. I'll also put you on the list to receive the coveted large format scan of the Philips 200 column cross-section poster (discussed earlier in this forum) which I should have constructed and cleaned up Real Soon Now.
Please contact me off-list. Thanks,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I'm trying to stain liver sections embedded in paraffin and fixed by metacarn fixative. My first antibody is monoclonal. The problem is that I only have a positive signal at the periphery of the section and nothing in the middle. Does someone have any clue why this could happen? Thanks in advance.
Prof. Dr. Francisco Javier Hernandez Blazquez Universidade de São Paulo Fac. de Medicina Veterinária e Zootecnia Departamento de Cirurgia - Setor de Anatomia Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
Thanks for the quick reply, Chris (and others who may be in the pipe),
Even my good luck seems to have a bad side! Not 5 minutes after posting this request, I found a copy of this paper, literally under my nose. Embarassing to say the least! I'll be quiet for at least a week....
Cheers,
Jim
Christopher F. Blanford wrote:
} James - } } Unfortunately we can't take any copies of the journal out of the } library. But as the RMS is just up the street from me, do you think } you could just give them a call? Either see if they have a copy they } could send you, or if you're still in a bind, see if they'd loan me a } copy for a day and I could scan in the pages you want. The number: +44 } 1865 248 768. } } Good luck. } } Chris } } On Wednesday, September 3, 2003, at 01:40 pm, James M. Ehrman wrote: } } } Hi all, } } } } I just about wrote "Urgent Assistance Needed" as the subject line, } } but then realized that would probably not make it past several dozen } } spam } } filters. I'm in my usual position (stuck between a rock and a } } deadline). I'm looking } } for a copy (original reprint or the journal issue) of: } } } } MANTON I. & VON STOSCH H.A. 1966. Observations on the fine structure } } of the male gamete of the marine centric diatom Lithodesmium } } undulatum. Journal of } } the Royal Microscopical Society 85: 119-134. } } } } I need to scan several figures from the paper, and our interlibrary } } loans can only } } get a xerox copy. Can anybody help? I'd gladly pay courier charges } } (both ways } } if you need the journal/paper back). Alternatively, if someone is } } willing, they could } } scan the figures from their end, but it would most likely be all of } } the plates in the paper, } } as I'm not sure right now which ones I need. } } } } If you help, I'll make several offerings on your behalf to the gods } } of microscopy, requesting } } long filament life, long strings of buttery smooth silver-gray thin } } sections, and awesome } } powers against the Dark Forces of Bureaucracy. I'll also put you on } } the list to receive } } the coveted large format scan of the Philips 200 column cross-section } } poster (discussed } } earlier in this forum) which I should have constructed and cleaned up } } Real Soon Now. } } } } Please contact me off-list. Thanks, } } } } 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/~jehrman } } } } } } } } } }
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (taylor-at-bio.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 11:21:33 ---------------------------------------------------------------------------
Email: taylor-at-bio.fsu.edu Name: Kenneth Taylor
Organization: Florida State University
Title-Subject: Fixation and Embedding of bacteria
Question: Hi,
I am looking for a protocol for fixing, embedding and sectioning bacteria, in particular E. coli. Good membrane preservation would be important for this study.
Does anyone have a protocol that they would be willing to share?
OK...thanks to all of the respondents about FESEMs. I did receive some good feedback.
I am trying to make a budget wedge for FY04 and need to know a basic idea of benefits of what I have selected as candidate systems. That said, I also need to put in funding wedges to cover the venue of appropriate SEMs.
JEOL 6700F Hitachi S-4800 Hitachi S-5200
Other options are welcome too.
I would appreciate knowing personal experiences based on differing sizes of specimens and different types of analysis. Budget time comes along one time, and is perversely ignorant of systems. Caveat emptor.
It may be time to upgrade. But to what? Cold FE with accommodation of my EDAX Cryospec is necessary. Robinson BSE is also a big plus.
Hello, could you please let me know who the suppliers of HISTORESIN are. It is a glycol methacrylate usful for light microscopy. It used to be marketed originally as LKB HISTORESIN. Thank you, Dobrusia
Dr Dobroslawa Budka Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa fax: +27-21-8433543 phone: +27-21-8431161 mobile phone: 0722656091
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reckert-at-nwlabs1896.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 21:24:36 ---------------------------------------------------------------------------
Question: We are interested in purchasing a CamScan CS44 SEM (appr. 10 years old) and I was wondering if anybody could share their experience (weak and strong points) about the scope. Thank you.
Well, BILLPC has stopped sending me copies of a virus, but I still get copies from PRIVAT (pD9E76707.dip.t-dialin.net [217.231.103.7], and MINIHOTEL (modemcable027.214-131-66.nowhere.mc.videotron.ca [66.131.214.27]
Some of the spoofed addresses/domains on the messages are ones I recognize from this list. So if any of you recognize these folks or their domains, would you please tell them to check out their machines. Thanks.
Hello Historesin is now produced and sold by Leica Microsystems Nussloch GmbH. Their address is Heidelberg Strabe 17-19 D-69226 Nussloch/Heidelberg Tel. 0 62 24/143-0 You may look at their homepage at http://www.histo-solutions.com/website/sc_hbu.nsf
Good luck
Prof. Dr. Francisco Javier Hernandez Blazquez University of São Paulo Fac. Veterinary Medicine and Animal Sciences Departament of Surgery - Sector of Anatomy Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "dobrusia budka" {dobrusiabudka-at-tlabs.ac.za} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 04, 2003 8:26 AM
Hi:
Help!!! I'm presently having problems with photography of nerve tissue. My problem is that the tissue appears shredded, and it has multiple holes and splits along the tissue but not the plastic. Making the tissue impossible to photo. This problem can only be seen when we are to photograph in the microscope. 1 micron light slides look fine. The only thing we have change is the type of fixative. We went from glutaraldehyde to para formaldehyde, but nothing have change from my protocol. We think is an infiltration problem, because the tissue is big to start and humidity have been a problem in the past in other types of tissue, and because the holes and splits are only in the tissue. We are looking for processing protocols for Sural Nerve tissue in order to compare with the one we have. If anyone can help it will be greatly appreciated.
Thanks
Omayra Velez MS Electron Microscopy Specialist Pathology Department Weill Cornell Medical College 1300 York Ave NY,NY, 10021 212-746-6437 omv2001-at-med.cornell.edu
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Does anyone know anything about the ISI WB-6 that is currently being offered on eBay? A friend of mine was wondering about the specifics concerning the condition and usability of the instrument.
Omayra It looks like fixation/impregnation problem. Paraformaldehyde itself is not enough for EM. I know nothing about your particular sample, in general, we are using 4% paraformaldehyde+ 1% GA overnight (+4oC). Nervous tissue also contains a lot of lipids, so dehydratation should be slow: actually, all steps should be longer than usual. I do find that Spurr resin is good for brain tissue: it penetrates better than Epon or Araldite. Good luck, Sergey.
At 02:04 PM 9/4/2003, 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 33-089 Los Angeles, CA 90095
Historesin as such is now discontinued but lives on as a series of resins produced by Heraeus Kulzer in Germany - Technovit 7100 (Historesin), Technovit 8100 (Historesin Plus) and Technovit 9100 NEW (Histodur).
These resins are all identical to the Historesin products. TAAB Laboratories Equipment Ltd is the Distributor for these products in the UK, but if anyone has a problem finding the distributor in their country, I may be able to help,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England Tel ++44 (0)118 981 7775 Fax ++44 (0)118 981 7881 e-mail sales-at-taab.co.uk www.taab.co.uk
I have problems with my EBSD measurements. The image of the phosphorscreen is distorted at the bottom of the screen while substracting the backgroung. instead of pure cirlces at the bottom you see more or less straight lines cutting the circles or the circles themselfs are distorted. When substracting at very low magnifiocations this problem doesnŽt occure. When you look at the RAW picture the area at the bottom shows a dark line. Please I need help. It is urgent
Regards Dirk Kirch -- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
I have problems with my EBSD measurements. The image of the phosphorscreen is distorted at the bottom of the screen while substracting the backgroung. instead of pure cirlces at the bottom you see more or less straight lines cutting the circles or the circles themselfs are distorted. When substracting at very low magnifiocations this problem doesnŽt occure. When you look at the RAW picture the area at the bottom shows a dark line. Please I need help. It is urgent
Regards Dirk Kirch
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
We are thinking to buy a liquid helium cooled double tilt TEM holder. Has anyone used this type of holder? Can you please tell me your experience with it? I would appreciate very much if you could tell me how well it works, and what problems it might have.
I would like to ask more specifically about the liquid helium cooling TEM holder made by USA side of the Gatan company. The UK made holder will be too expensive for us, so anyone has experience with the US made holder?
Hi to all: Thanks for the responce on the nerve problem. Some of you have asked how big the tissue samples are and the type of resin I'm using. We are using, EMBED 812 with DMP-30 as catalyst. The tissue size is about 5mm x 2mm x 3mm. Big for the most part. I already tryed to make the blocks smaller, but the nero-pathologist wanted the pieces this big. We have a 20 hour protocol from buffer to 100% epon. I'll try some of the sudjestions, lenthtening the protocol. Although we think we are also having a humidity issue as well, due to construction and AC problems.
Thanks again
Omayra Velez Electron Microscopy Specialist Pathology Department Weill Cornell Medical College 1300 York Ave NY,NY. 10021 212-746-6437 omv2001-at-med.cornell.edu
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Here is the table of contents for the September/October 2003 issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 9 September for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting. Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA. MSA members anywhere have free subscriptions. Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
Table of Contents:
Carmichael Print Your Own Organs! Kelly and Gribb Atom Probes LEAP Ahead Breger Microscopy At The Ends Of The Earth Barkan, et al. A Si Multi-Cathode Detector For Microanalysis Applications Sehgal, Karim, Stafford, & Fasolka Techniques for Combinatorial and High- Throughput Microscopy Part 1: Gradient Spec. Fabrication for Polymer Thin Film Research Millette, Boltin, Few, & Turner Microscopical Studies of World Trade Center Disaster Dust Particles Vrdoljak Prep. Of Soil Samples For Light And TEM Radzikowska A New Look At Cast Iron Microstructure Lifshin & Gauvin Precision and Detection Limits for EDS Analysis in the SEM Sedgewick Adding Color To Grayscale Images Downing Support Films with Uniform Hole Size Mills Cleaning a Cold Cathode Gauge Tube Yang and Kalab Zigzag Edges in SEM Micrographs Schooley Microscopy CD-ROMs For Children: A Bibliography
Does anyone have a package or know of a package that can calculate changes in sample thickness from thickness fringes observed in conventional TEM? I am imageing defect free single crystals on the order of 100nm. I need to know how the crystals thickness profile changes.
I have a student who wants to image magnetic particles with SEM. The particles are in the 400nm to 1µm size range. I had planned to stick them to double stick copper tape prior to imaging. However, I am concerned about their adversely affecting the microscope. Are there any suggestions as to mounting the particles, whether short working distance may magnify potential problems if some particles break loose and get into the lens. Etc?
If there is a concern then I guess it would be possible to mount on sticky carbon tape, overlay the sample with a formvar film, and then use backscattering to image the particles. However, is this necessary?
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
If one wants to do pure standards-based quantitative EDS analysis, protocol says that conditions for the standards must be the same as for the specimen. Well, if one is using a thermal FE SEM, then the beam current is very likely to be the same at any time. How about for a cold FE gun?
From what I can surmise, the cold guns vary much more than thermal FE guns, to the point that intervention is necessary. Does this mean that every quant EDS measurement has to have a precursor Faraday cup reading of beam current? Conversely, does this mean that this is not required when using a thermal FE gun?
The EDAX system supports a Beam Current Factor so that calculations are made based on as-read current when making the quant analysis. This seems like a lot of hassle. Is the difference between standardless and standards-based EDS so huge that the hassle is warranted? Or, is this nit picking?
Is it possible to simply degauss the samples before putting them in the SEM?
There is a border, I believe, between too much degaussing and too little. Looking at hard drive tracks is a case in point. Depending on your ultimate resolution, the working distance may facilitate your specimens by keeping them away from the final lens. 1um specimens should not require short WD.
Your BSE ought to be a separate issue depending on what type it is and what KV it works at.
gary g.
At 03:14 PM 9/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have routinely examined (gamma-)ferric oxide particles in the SEM by dusting the particles onto double-sided sticky carbon tape. Or you can paint a small dab of carbon paint onto your stub and dust the particles onto the paint while it is still wet. Then tap the side of the stub firmly on a hard surface to dislodge any loose particles. If you are really paranoid you can use a low-pressure gas stream (like a "Micro Duster") to make sure there are no loose particles present. However, the more aggressively you remove looser particles, the less you are likely to see larger particles that may exist in the specimen. This may be important if you are trying to do particle size distribution measurements.
Particles like Fe2O3 are weakly magnetic, so there is not much danger of them being sucked up into your lenses by the magnetic field.
You will probably want to keep the working distance fairly short ( {10mm) to achieve acceptable image quality.
Hope this helps, Ken Gaugler
Debby Sherman 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 have a student who wants to image magnetic particles with SEM. The } particles are in the 400nm to 1µm size range. I had planned to stick them } to double stick copper tape prior to imaging. However, I am concerned about } their adversely affecting the microscope. Are there any suggestions as to } mounting the particles, whether short working distance may magnify potential } problems if some particles break loose and get into the lens. Etc? } } If there is a concern then I guess it would be possible to mount on } sticky carbon tape, overlay the sample with a formvar film, and then use } backscattering to image the particles. However, is this necessary? } } 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 } } }
Wouldn't it also be dependent on the type of SEM used (i.e. HRSEM)?
If the final lens (pole piece) is not energized for through-the-lens detection, the sticky Cu tape should hold them down. Disperse the particles then blow off the weakly held particles. Now lower a ferro magnetic material over the samples and see if any pop off. If particles do "pop" off then use formvar and a high kV. The magnetism of the particles may play a role in their physical construct so degauss with caution. I believe imaging aberrations will most likely be the issue over that of tool contamination. I have successfully imaged toner particles in a relatively low resolution SEM. It should be noted that this tool has been classified as a "dirty" tool. The toner particle were even dispersed on the "new" C sticky tabs. The ones that are rock hard with barely any adhesive.
Kristopher
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, September 05, 2003 7:28 PM To: Debby Sherman Cc: MSA listserver
Is it possible to simply degauss the samples before putting them in the SEM?
There is a border, I believe, between too much degaussing and too little. Looking at hard drive tracks is a case in point. Depending on your ultimate resolution, the working distance may facilitate your specimens by keeping them away from the final lens. 1um specimens should not require short WD.
Your BSE ought to be a separate issue depending on what type it is and what KV it works at.
gary g.
At 03:14 PM 9/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You really don't need a package. You can do this by hand using basic TEM techniques.
I think that your best way to do this would be to set up a good 2-beam condition and use convergent beam electron diffraction. You use the distances between the symmetrical fringes (Kossel-Mollenstedt fringes) in the known reflection, calculate the deviation from exact Bragg condition for each fringe, and plot the results assuming that the first fringe that you see is the 1st, 2nd, or 3rd. You plot s(i)^2/n(i)^2 vs 1/n(i)^2, where n(i) is the index of the particular fringe. You try different starting values for n(1) starting from 1, then 2, then 3. You usually don't have to go higher than 3. When you get a straight line, the slope of the straight line is -1/(extdist)^2 and the intercept of the line is 1/t^2. This comes from the following relationship,
s(i)^2/n(i)^2 + 1/(extdist)^2/n(i)^2 = 1/t^2.
This technique is demonstrated in a number of TEM texts. The procedure is written in the Williams and Carter book.
You could apply this at two points on the sample along the wedge and get the angle.
You could also do this from the results of the two-beam approximation model. If you are at 2-beam Bright Field condition and you do not have bend contours present, then the first dark fringe that occurs in the wedge will occur at a thickness of 0.5 * (extdist(g)) and the second at 1.5 * (extdist(g)), (and so forth.) From the lateral separation distance between the fringes and the value of the extinction distance for that reflection, you can calculate the wedge angle. This is also written up in the Williams and Carter book and others as well.
-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: thurston e herricks [mailto:thurst0n-at-u.washington.edu] Sent: Friday, September 05, 2003 3:37 PM To: Microscopy-at-sparc5.microscopy.com
Hello all,
Does anyone have a package or know of a package that can calculate changes in sample thickness from thickness fringes observed in conventional TEM? I am imageing defect free single crystals on the order of 100nm. I need to know how the crystals thickness profile changes.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (taylor-at-bio.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 11:21:33 ---------------------------------------------------------------------------
Email: taylor-at-bio.fsu.edu Name: Kenneth Taylor
Organization: Florida State University
Title-Subject: Fixation and Embedding of bacteria
Question: Hi,
I am looking for a protocol for fixing, embedding and sectioning bacteria, in particular E. coli. Good membrane preservation would be important for this study.
Does anyone have a protocol that they would be willing to share?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Jlindstrom-at-Hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, September 2, 2003 at 17:19:35 ---------------------------------------------------------------------------
Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (icastro-at-usiminas.co.br) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, September 8, 2003 at 09:26:44 ---------------------------------------------------------------------------
Email: icastro-at-usiminas.co.br Name: Ivan de Castro
Organization: Usiminas
Education: Graduate College
Location: Brazil
Question: I'm having some difficulty on metallographic examination of Hot Dip Galvanized steels. Then I will shall be pleased if you could help me giving some information about papers that discrebe the better technique to metallographic prepation and etching of HDG steels. Thanking you Yours sincerely
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Jlindstrom-at-Hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, September 2, 2003 at 17:19:35 ---------------------------------------------------------------------------
Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
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Email: maloneyb-at-fiu.edu Name: Barb
Organization: FIU
Title-Subject: Receipe for cultured cells on filter for SEM
Question: Dear Group - what is the newest and most successful receipe for prep for SEM on cultured cells on a filter? Thanks Barb
The server is back up, albeit running slowly due to what I believe are DNS problems.
If you are having problems posting, please let me know so that I can investigate. Don't be bashful to forward to me (complete) copies of any error messages.
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Email: babbini-at-units.it Name: Lorenza Babbini
Organization: Dept. of Biology - University of Trieste - Italy
Title-Subject: MListserver:
Question: Hi all! I have to do my sections of decalcified coralline algae and I would like to try to use Histocryl resin. Is there anyone that ever used it for sectioning plants? Suggestions and comments are welcome! Lorenza
Does anyone have for sale a used but still properly functioning Metaltek filter wheel, Model # 5240? The wheel itself is all we'd need, but the controller could be useful as well. Or possibly a Uniblitz shutter model # V505S1Z0? The shutter in our Metaltek has blitzed its last, and I am suspicious of the wheel itself as well. Apologies to those who get this message twice from both the microscopy and the confocal lists.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
I have a 1950's vintage AO Spencer microtome with the porro prism, binocular head. In the past, we've had a stereomicroscope with the same head and the same problem, which is prisms breaking loose from the adhesive mounting. I know that it's next to impossible to try to achieve freehand alignment when re-cementing, and am told that there's a jig for this purpose. Not having the jig, and imagining that those who would posess one and the know-how to use it are superannuated, if still living, I'm turning to the group at large for any helpful leads in getting this instrument repaired. Or, does anyone have the heads with intact prisms that you would be willing to sell? Or, am I better off just getting another, used microtome altogether?
Much obliged,
Dave Stadden Research Scientist Testing & Analysis Lab Armstrong World Industries, Inc.
I really do have some worthwhile information to offer, but the short answer may be, it all depends.
The importance and frequency of beam current measurements will depend on the stability of your beam and your tolerance for error. If your beam current only fluctuates by a percent or so over a few minutes, you might be able to ignore it. The uncertainty in EDS results is going to be a larger error than the relative error from beam drift. However, if you experience 5% drift in 5 minutes, I would definitely recommend frequent checks of the beam current to correct for the drift. I say this not having worked with FE guns, so I don't know how much drift to expect. I definitely like to have current measurements often enough so that there is no more than a 5% change in beam current between measurements. More critical measurements would necessitate even more frequent checks.
The type of quant analysis will also make a difference. If you are monitoring all elements in the sample you can normalize your results which should cancel out the effects of beam drift. At moderate count rates all elements should be affected similarly, i.e., only the intensity should drop. The change should not affect the matrix correction, if the software is written correctly. You could check your software by forcing a major change in beam current via the condenser lens to see if the results are the same.
However, if you are not directly quantifying all elements present, e.g., you are analyzing an element by difference, then an accurate measure of beam current will be important. Whatever loss (or gain) in current you encounter will be attributed to the element determined by difference. If that element is not critical (e.g., C in a polymer), then you only need be concerned about the loss (or gain) of concentration in proportion to the current drift. But if you are trying to determine a small component (e.g., 5% Li by difference) it will be profoundly influenced by the beam drift.
The difference in standardless analysis and analysis with your own standards will depend on your system and its software. Some systems use built-in standards for their "standardless" analysis. Those built-in standards are probably collected on another scope in another lab under different conditions, but they can produce some pretty good results when corrected for current scope conditions.
If your technique is good, you might be able to improve on the "standardless" results by collecting your own standards. For instance, our internal standards were collected at the highest resolution mode on our analyzer while we typically do our x-ray work at a faster mode to improve statistics. Therefore, we can improve our deconvolution and thus our results by collecting our own standard profiles. You just need to be sure that your techniques are as good or better than those of the application engineers for your EDS system. I know it took me a while to relearn the lesson that the carbon coat on the standards and unknowns must be identical. The coat can cut the intensities by a few percent.
My personal experience is that when mixing line series (K, L, and M), the use of my own standards usually leads to a substantial improvement in accuracy. For analyses within the line series, I find that I cannot necessarily improve the results.
So like I said, "it depends" whether the close monitoring of beam current will be important or not.
HTH, Warren
At 04:45 PM 9/5/2003 -0700, you wrote:
} Greetings to EDS experts: } } If one wants to do pure standards-based } quantitative EDS analysis, protocol says } that conditions for the standards must } be the same as for the specimen. Well, } if one is using a thermal FE SEM, then } the beam current is very likely to be } the same at any time. How about for a } cold FE gun? } } From what I can surmise, the cold guns } vary much more than thermal FE guns, to } the point that intervention is necessary. } Does this mean that every quant EDS } measurement has to have a precursor } Faraday cup reading of beam current? } Conversely, does this mean that this is } not required when using a thermal FE gun? } } The EDAX system supports a Beam Current } Factor so that calculations are made based } on as-read current when making the quant } analysis. This seems like a lot of hassle. } Is the difference between standardless and } standards-based EDS so huge that the hassle } is warranted? Or, is this nit picking? } } gary g.
------------------------------------------- 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
Arthur Rosenfelder roseoptics-at-aol.com has the tools to rebuild these scopes or can probably swap out on that is already reconditioned.
Rosenfelder Optics, Specializing in Microscope Service and Repair. Maintaining research and instructional optics since 1980, P.O. Box 164, Bellvue, Colorado 80512, (970) 217-3368 RoseOptics-at-aol.com Art often has microscopes as well as parts and repair for sale email him for you needs. Art also remounts and realigns prisms in AO Spenser Cycloptic stereo microscopes.
Gordon Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. ----- Original Message ----- } From: "David R Stadden" {DRStadden-at-armstrong.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, September 11, 2003 10:33 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (halbers-at-doacs.state.fl.us) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, September 11, 2003 at 14:55:45 ---------------------------------------------------------------------------
Email: halbers-at-doacs.state.fl.us Name: Susan Halbert
Organization: Division of Plant Industry, Florida Department of Agriculture
Education: Graduate College
Location: Gainesville, FL USA
Question: I have a late '70s vintage Zeiss universal microscope with a built-in camera. Some time ago, the camera blew a fuse. We replaced the fuse, and it blew again, indicating an electrical malfunction. We have not been able to find anyone that can fix it. Does anyone have any suggestions?
Dear All, I would like to post the following question: Is there a sensitive method to measure the intracellular pH by a staining procedure of tissue sections (pancreas tissue)? Thanks for any suggestions! Daniel
-- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V= Developmental Biology & Molecular Pathology University of Bielefeld D 33501 Bielefeld/Germany
By the time you fix the tissue, embed in paraffin (or other medium) and section, then the living intracellular pH is longsince gone. However, some work has been done on living cells in culture, using special fluorescent dyes that change their color or density at different pHs. I don't have any references handy, but you could probably find some by searching PubMed or Google.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Friday, September 12, 2003 1:22 PM +0200 DANIEL EBERHARD {daniel.eberhard-at-uni-bielefeld.de} wrote: } } Dear All, } I would like to post the following question: } Is there a sensitive method to measure the intracellular pH by a staining } procedure of tissue sections (pancreas tissue)? Thanks for any } suggestions! Daniel } } -- } (-)-(-) } --------------------------- \"/ --- } Dr. Daniel Eberhard =V= } Developmental Biology & Molecular Pathology University of Bielefeld } D 33501 Bielefeld/Germany } } FAX: xx49(0)521-106-5654 (-)-(-) } --------------------------- \"/ --- } =V= } }
Hello Daniel, There are a number of fluorescent dyes which can be used to determine intracellular pH, the most useful variants exhibit shifting of the emmission spectra with changes in pH. You can check Molecular Probes' website for more information: http://www.probes.com/servlets/directory?id1=3&id2=76 -Karl G.
DANIEL EBERHARD 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 } ----------------------------------------------------------------------- } } Dear All, } I would like to post the following question: } Is there a sensitive method to measure the intracellular pH by a } staining procedure of tissue sections (pancreas tissue)? } Thanks for any suggestions! Daniel }
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
I would be grateful for any input from the listserve for the following request I recently received. Rosemary
"I am working on a crime scene investigation outreach activity for 4th-7th graders. I have the activities prepared already, but I would like to include more hard science involving fiber analysis and fingerprinting. I am familiar with several types of microscopy, but I was wondering if you could help me with the explanations of their application to forensic science. We are also hoping to be able to put images on the outreach website and the final paper version so that students could see, for instance, the differences between types of fibers and have an idea of how a match is made."
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 Friday, September 12, 2003 at 14:02:21 ---------------------------------------------------------------------------
Email: hgabrisc-at-uno.edu Name: Heike Gabrisch
Organization: University of New Orleans
Title-Subject: MListserver: negative scanner
Question: Can anybody recommend a good scanner for TEM negatives?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gilweiss-at-worldnet.att.net) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, September 12, 2003 at 19:44:55 ---------------------------------------------------------------------------
Question: What is the practical use of circular polarization in microscopy? What physical phenomena initiate it? I have not found a good working source of info on the subject.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gilweiss-at-worldnet.att.net) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, September 12, 2003 at 19:44:55 ---------------------------------------------------------------------------
Question: What is the practical use of circular polarization in microscopy? What physical phenomena initiate it? I have not found a good working source of info on the subject.
There are scanners available over a wide price range. I bought one a few years ago, on a tight budget, and I'm happy with it. Its an Epson Expression Pro 1600. I came with a trans-illuminator for transparencies and runs with Photoshop. I use 400 dpi for "work print" type images, and a higher dpi for publication. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I´d like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. I´d like to have a high resolution. Could someone give me a suggestion about the best device (as brand) to apply? Thank you. Best Regards, Massimo
I´d like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. I´d like to have a high resolution. Could someone give me a suggestion about the best device (as brand) to apply? Thank you. Best Regards, Massimo
On Friday, September 12, 2003, at 08:06 PM, by way of Ask-A-Microscopist wrote:
} Name: Gilbert Weiss } } Organization: Imaging Analysis } } Education: Graduate College } } Location: Stamford, CT } } Question: What is the practical use of circular polarization in } microscopy? What physical phenomena initiate it? I have not found a } good working source of info on the subject. } } Thanks for the help. } Dear Gilbert, I'll let the LM experts answer your first question. As for the second, molecules that do not have a center of symmetry will interact differently with right-hand and left-hand circularly polarized light. The usual description is to consider a molecule that has a carbon atom singly bonded to four distinct groups, such as H, F, Cl, and Br. There are two possible configurations, which cannot be superimposed, but which are mirror images of each other. (By rotating any configuration it can be superimposed onto one of the two possible configurations.) Any optics text should have a description of this phenomenon, so I am surprised you have not found one. Yours, Bill Tivol 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
I have a Zeiss Confocal LSM-510 for sale which has the Argon Helium lasers and I am looking to sell it.
Photos and a complete configuration are available by request. It is located in Bohemia NY and is able to be inspected with an appointment.
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
Is anyone aware of the fluorescent properties of 5-(4-dimethylaminobenzylidene)rhodanine....not rhodamine. This is a histochemical stain fro copper usually used for brightfield microscopy, but I am also seeing a great fluorescent signal. Thanks, Brett
Brett M. Connolly, Ph.D. Merck & Co.,Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly-at-merck.com
------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (Whitehouse Station, New Jersey, USA), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD) that may be confidential, proprietary copyrighted and/or legally privileged, and is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please immediately return this by e-mail and then delete it. ------------------------------------------------------------------------------
"WebCam" and "High Resolution" are generally mutually exclusive. The required bandwidth for sending high res video over the Internet (and many "internal" networks) makes it unworkable. As a result, depending on what you're after, you may need two cameras. (Even many "high end" video cameras don't have a very high resolution, at least as compared to "still" digital cameras.)
WebCams, because of the low resolution are available from a number of companies, and there isn't much difference between them. They all offer at least a couple of models. The differences tend to be in zoom, microphone, and the level of "remote control". It may take some ingenuity to get one of these mounted.
As far as High Res, that subject comes up often in this group, and the Nikon CoolPix line seems to be most popular. There are microscope adapters available for the Nikon's and some of the other more popular choices.
Thank you;
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
-----Original Message----- } From: max_gra-at-libero.it [mailto:max_gra-at-libero.it] Sent: Monday, September 15, 2003 8:34 AM To: MSA Microscopy
Hi all,
I´d like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. I´d like to have a high resolution. Could someone give me a suggestion about the best device (as brand) to apply? Thank you. Best Regards, Massimo
Your question is not only a good one, it offers many people a new way do image analysis, strangely enough!
We have to start with regular polarized light. The difference between "ordinary light" and "polarized light" has to do with how the electric vectors are vibrating. Because light is electro-magnetic radiation, it has both an electrical component and a magnetic component. Most microscopists are only interested in the electrical function.
We often use a sine wave to describe light. The sine wave is actually the path traveled by the tip of light's electric vector as it builds up in a positive direction, then decreases, then builds up in a negative direction then decreases. This process gives light several key features: a. a direction of travel (you see an object because light is traveling from it to you) b. a direction of vibration (the direction in which the vector increases and decreases; direction of vibration is always perpendicular to the direction of travel) c. its intensity (intensity is proportional to the square of the maximum displacement in a positive or negative direction) c. its wavelength (which microscopists typically associate with color... all apologies to the physicists in the audience).
Ordinary light is comprised of waves of light traveling toward you, vibrating at all angles. The typical analogy: if you looked back along the direction of travel, you would see all the vectors like arrows vibrating N-S, E-W, and all angles in between.
To convert ordinary light to polarized light, you need to have it interact with something which preferentially absorbs all but one direction of vibration. That "something" could be the process of reflection at the specific angle known as "Brewster's angle" (dependent on the properties of a given material), absorption through something like a polarizing filter or polarizing sunglasses, or the process of beam splitting which happens when the beam enters a material which is birefringent. As this term implies, these materials have two ("bi") characteristic refractive ("refr") indices ("in"). (The "gent" means "having the property of..."). The refractive indices are the result of differences in electric field which result from the arrangement of atoms, ions, or elecrons. In the simplest term, birefringence is the electrical equivalent of wood grain: one direction is "with the grain" and the direction at right angles is "against the grain". Many minerals are birefringent, su ch as calcite and quartz. Also, many polymer fibers and films.
If you create polarized light by beam splitting, the beam will be split into two components: an "e" ray and an "o" ray. They will have directions of vibration at right angles to each other. Each will encounter a different electrical field. Both will slow down because of the interaction between the electrical field in light and the field in the object. The one which encounters the field represented by the higher refractive index ("against the grain") will slow down more and lag behind its partner. The "lag" is called "retardation" and is measured in wavelengths.
So, what does all of this have to do with circularly polarized light? This part is really tough without diagrams, but draw the following: A horizontal reference line On the left side of the line, draw rectangle to represent a polarizer which permits light to emerge vibrating at +/- 45 degrees = Electric Vector "E" At some distance along the line, draw another rectangle to represent a piece of birefringent material (ex: a polymer overhead transparency) In that rectangle, draw a short vector from the reference line pointing up (we'll call that "e") and another, again from the reference line, at 90 degrees ("o"). At a third point along the reference line, draw another short vector "e" and, a short distance further along, draw another "o".
Here is the explanation: a. When ordinary light passes through the polarizing filter, it absorbs all the electric vectors except "E". (We would have determined the direction of vibration by rotating the filter). b. When the polarized beam enters the birefringent material, it is split into "e" and "o", each polarized and each vibrating at right angles to each other. c. Because each interacts with a different electrical field, one will slow down and lag behind the other. In this case, we have drawn the example when "e" experiences the greater interaction and, therefore, lags behind "o". d. Analyzing the resultant electric vector will tell you the "State of Polarization": Again, a simple drawing helps. Draw two perpendicular lines which cross in their centers. Label the "Y" with + and - (top and bottom); Label the "X" with + and - (right and left). The next part takes a bit of imagination (there is a great diagram in Optimizing Light Microscopy, but I can't draw it here). First, keep in mind that "e" and "o" are vibrating at right angles. Let's say "e" is vibrating in the Y direction and "o", in the X direction. If "e" lags behind "o" by some multiple of whole ("e" goes to max in + direction when "o" goes to max in + ) or half wavelengths ("e" goes to max in + when "o" goes to max in -), the resultant vector would see-saw between +45 and -45 degrees, generating "linearly polarized light". (Direction changes, depending on whether a whole wave or half wave) If "e" lags behind "o" by odd multiples of quarter waves, the resultant vector travels in a corkscrew pattern. Quarter wave shifts produce clockwise rotation; three-quarter shift produces counter-clockwise rotation. This corkscrew path is what we call "circularly polarized light". Circular polarizing filters are actually sandwiches of (a) a regular polarizing filter and (b) a piece of plastic which causes a retardation of 1/4 wave (typically about 140nm).
Why is all this important? And what does it have to do with image analysis? The biggest challenge in image analysis is simplifying objects so that they can be recognized by an automated measuring system. In polarized light, birefringent objects viewed between crossed polars are bright against a dark background ... great for segmenting with an automated system. Unfortunately, the intensity changes with orientation (yet another discussion). The same crystal, for example, will appear dark if sitting N-S or E-W but bright at +/- 45 degrees! On the other hand, if you use crossed CIRCULAR polars (make sure the retarding plastic is toward the center of the "sandwich" and the sample is in between), all the orientation effects will be removed because of the corkscrew path of the light. The result: all birefringent objects which are behaving the same would appear the same intensity, regardless of orientation.
We used this trick very successfully, for example, with the Center for Urban Ecology, when they were trying to measure the % crystallinity of soil samples.
Note: if you are interested in trying circularly polarized light, you can ask your microscope rep to lend you a set of circular Pol filters or you can make your own, using 2" squares of polarizing filter and "quarter wave" material, typically available from companies like Edmund Scientific. (Caveat: no financial interest).
Whew! A long winded answer, but I hope that it is helpful.
If you are interested in copies of the diagrams described above, contact me off line.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
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I am using Luft's technique for staining the glycocalyx with Ruthenium Red. I know the RR is supposed to be included in the aldehyde fix, the rinses and the osmium but how about the osmium rinses or ethanol dehydration series? Has anyone successfully done this procedure? Thanks, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Don't bother even trying a webcam, they have insufficient resolution (unless all you want to do is to obtain images small enough to stream on the Net).
I've been there.
Buy a Nikon Coolpix 4500 or similar plus an adaptor from any of several vendors. I bought my adaptor from WPI (www.wpiinc.com).
It works well.
cheers
rtch
Date sent: Mon, 15 Sep 2003 15:34:03 +0200
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmolin-at-cooksonelectronics.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, September 15, 2003 at 13:43:45 ---------------------------------------------------------------------------
Email: dmolin-at-cooksonelectronics.com Name: Dick Molin
Organization: Cookson Electronics
Education: Graduate College
Location: Indianapolis, IN USA
Question: Our company specializes in applying a thin, transparent conformal coating called Parylene. I'm wondering if the use of DIC could be helpful in my analysis of the coating and the coating/substrate interface. I realize DIC is valuable for transparent biological specimens,and don't know if this usefulness translates for my application. Please let me know your opinion since I would want to justify the added cost for the prism and the optics.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 15, 2003 at 15:36:08 ---------------------------------------------------------------------------
Email: echeung-at-eyetk.com Name: Eunice Cheung
Organization: Eyetech Pharmaceuticals
Title-Subject: MListserver: Trouble with Formvar and glass coverslips
Question: I am doing a project in growing cells on Formvar covered grids. First, the film is made by floating the Formvar on water. Second, the 7 grids are put on the film. Third, a small circular glass coverslip is place covering on top of the grids. Fourth, the formvar, grids, and coverslip are picked up with a piece of parafilm. Afterwards, the coverslip is taken out with the grids in place and sterlized in UV. Later it is coated with gelatin. The problem: The Formvar with the grids falls off from the coverslip. Any suggestions in preventing this? Thanks.
I am looking for people who have equipment to give away such as micrscopes that they would not mind parting with, that would serve as a teaching tool for high school students from grades 7 to 11. The ideal microscopes would be with trinocular tubes or without. I would also welcome video or digital cameras. Video microscopy would be a great help in teaching large groups of kids. Any other laboratory or imaging equipment or supplies would also be welcome.
Hi, You can do a good job with negative staining. Best regards from Prague Oldrich On 15 Sep 2003 at 11:50, Jon Krupp 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: } } Is there a quick and dirty method for looking at TMV? } } I have never done it, a person from the local community wants to see if } that's what killing his plants and insists on 'seeing' it with his own } eyes. } } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
you haven't mentioned the size of your TEM negatives but I will assume that they are one of the larger formats (eg 83x84mm or 83x102mm).
I have used an Epson Perfection 1200 which produce fairly good results but I was always suspicious of its quoted top resolution of 1200 dpi. Negatives on this type of flatbed scanner have to be placed on the glass surface of the scanner to be scanned which creates several problems: Newton's rings; dust and other marks on the glass.
I now use a Microtek Scanmaker 8700 which you can buy with or without Silversoft image management software (I think its the PRO version in the US). It also has a glass panel for normal flatbed document scanning but importantly has a separate glassless drawer which can take several types of film holder. See website below: http://www.microtekusa.com/sm8700.html So far I have been very impressed because the scan quality is good and some of our staff have even used the 35mm film and slide adaptors to digitise their colour slides (1200dpi is adequate for 35mm if you're using a digital projector or displaying on screen). This model comes with USB 1.1 but more usefully Firewire with an adaptor card. In the UK you also get Adobe Photoshop Elements which is quite handy. There is sufficient room to place two large 83 x 102mm (3 1/4 x 4 inch) in the glassless film carriers. Although I am still just using the 5 x 4 inch adaptors I suspect that it would be possible to construct inserts.
There seem to be 3 main types of possible scanner: 1. Dedicated film scanners - these are often the best and highest resolution but large format film scanners are extremely expensive and rare (1,000 - 2,000 UK pounds and upwards). Make sure it really will do your film size. 2. Normal flatbed document scanners with a glass copying surface and a film/transparency adaptor light source either built into the lid of the scanner or replacing the lid. These can be extremely cheap with current resolutions in excess of 1200dpi but be sure that the transparency adaptor is not just for 35mm or medium format (costs start from below 100 UK pounds). A good way of trying out but the quality won't be as good as the other two types. 3. The hybrid flatbed scanner with separate glassless film carriers which slide into a drawer in the scanner body. These seem to strike a good balance in terms of quality, versatility and cost for large format film (600 UK pounds and upwards).
Basic things to look for are a genuine 1200 dpi or higher resolution (after all you may want to enlarge part of an image or examine in detail on the screen). Colour bit depth of 42 or 48 (ie 14 or 16 in greyshades). The ability to generate a variety of image formats (especially TIF, BMP and JPEG). If you have a lot of dense negatives check the D max or dynamic range - it should be at least 3.4 but 4.0 would be better for difficult film. Finally the connection to the computer should really be USB 2.0 or Firewire because if you scan a 4x3 inch negative at 1200dpi that's 17280000pixels (ie 17-18 Mb minimum for 8 bit B&W) which can be quite slow on USB 1.1.
Most e.m. film I scan is at 600 or 800dpi 8 bit grey-scale and stored as best quality JPEG. TIF and 14/16 bit greyscale would be nice but in the real world I still have the negatives and can store more digital images on CDs or hard disk. I can't remember the last time I produced an old style photographic print, although I do still offer the option for final publication quality
I hope this helps and if the 8700 isn't good enough I am sure either Microtek or a competitor will do higher resolution or better D max.
By the way if anyone has read this far I am seriously considering the Canon i950 bubble/inkjet printer as a successor to our Epson 750. Has anyone used the Canon for e.m. black and white printing because it is supposedly quiet, good resolution, faster and has a separate replaceable print head?
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK tel no: 0191 515 2872 / 3468 e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: hgabrisc-at-uno.edu (by way of Nestor J. Zaluzec)
Hello everyone,
If possible, I would like to find out the hourly rate for SEM/EDS work. Our management wants to increase the rate and we are trying to asses the "fare" rate for SEM work. I would like to find out the rate that you would charge and any other charges (extra copies, digital, WDS work, etc)
Thank you for the info.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
An enlightening discussion, as usual! Thanks for taking the time to put that all down in electrons!
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Barbara Foster {bfoster-at-mme1.com} 09/15/2003 08:50 PM
To: microscopy-at-msa.microscopy.com cc: Subject: Re: Ask-A-Microscopist: circular polarization in microscopy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Gilbert,
Your question is not only a good one, it offers many people a new way do image analysis, strangely enough!
We have to start with regular polarized light. The difference between "ordinary light" and "polarized light" has to do with how the electric vectors are vibrating. Because light is electro-magnetic radiation, it has both an electrical component and a magnetic component. Most microscopists are only interested in the electrical function.
We often use a sine wave to describe light. The sine wave is actually the path traveled by the tip of light's electric vector as it builds up in a positive direction, then decreases, then builds up in a negative direction then decreases. This process gives light several key features: a. a direction of travel (you see an object because light is traveling from it to you) b. a direction of vibration (the direction in which the vector increases and decreases; direction of vibration is always perpendicular to the direction of travel) c. its intensity (intensity is proportional to the square of the maximum displacement in a positive or negative direction) c. its wavelength (which microscopists typically associate with color... all apologies to the physicists in the audience).
Ordinary light is comprised of waves of light traveling toward you, vibrating at all angles. The typical analogy: if you looked back along the direction of travel, you would see all the vectors like arrows vibrating N-S, E-W, and all angles in between.
To convert ordinary light to polarized light, you need to have it interact with something which preferentially absorbs all but one direction of vibration. That "something" could be the process of reflection at the specific angle known as "Brewster's angle" (dependent on the properties of a given material), absorption through something like a polarizing filter or polarizing sunglasses, or the process of beam splitting which happens when the beam enters a material which is birefringent. As this term implies, these materials have two ("bi") characteristic refractive ("refr") indices ("in"). (The "gent" means "having the property of..."). The refractive indices are the result of differences in electric field which result from the arrangement of atoms, ions, or elecrons. In the simplest term, birefringence is the electrical equivalent of wood grain: one direction is "with the grain" and the direction at right angles is "against the grain". Many minerals are birefringent, su ch as calcite and quartz. Also, many polymer fibers and films.
If you create polarized light by beam splitting, the beam will be split into two components: an "e" ray and an "o" ray. They will have directions of vibration at right angles to each other. Each will encounter a different electrical field. Both will slow down because of the interaction between the electrical field in light and the field in the object. The one which encounters the field represented by the higher refractive index ("against the grain") will slow down more and lag behind its partner. The "lag" is called "retardation" and is measured in wavelengths.
So, what does all of this have to do with circularly polarized light? This part is really tough without diagrams, but draw the following: A horizontal reference line On the left side of the line, draw rectangle to represent a polarizer which permits light to emerge vibrating at +/- 45 degrees = Electric Vector "E" At some distance along the line, draw another rectangle to represent a piece of birefringent material (ex: a polymer overhead transparency) In that rectangle, draw a short vector from the reference line pointing up (we'll call that "e") and another, again from the reference line, at 90 degrees ("o"). At a third point along the reference line, draw another short vector "e" and, a short distance further along, draw another "o".
Here is the explanation: a. When ordinary light passes through the polarizing filter, it absorbs all the electric vectors except "E". (We would have determined the direction of vibration by rotating the filter). b. When the polarized beam enters the birefringent material, it is split into "e" and "o", each polarized and each vibrating at right angles to each other. c. Because each interacts with a different electrical field, one will slow down and lag behind the other. In this case, we have drawn the example when "e" experiences the greater interaction and, therefore, lags behind "o". d. Analyzing the resultant electric vector will tell you the "State of Polarization": Again, a simple drawing helps. Draw two perpendicular lines which cross in their centers. Label the "Y" with + and - (top and bottom); Label the "X" with + and - (right and left). The next part takes a bit of imagination (there is a great diagram in Optimizing Light Microscopy, but I can't draw it here). First, keep in mind that "e" and "o" are vibrating at right angles. Let's say "e" is vibrating in the Y direction and "o", in the X direction. If "e" lags behind "o" by some multiple of whole ("e" goes to max in + direction when "o" goes to max in + ) or half wavelengths ("e" goes to max in + when "o" goes to max in -), the resultant vector would see-saw between +45 and -45 degrees, generating "linearly polarized light". (Direction changes, depending on whether a whole wave or half wave) If "e" lags behind "o" by odd multiples of quarter waves, the resultant vector travels in a corkscrew pattern. Quarter wave shifts produce clockwise rotation; three-quarter shift produces counter-clockwise rotation. This corkscrew path is what we call "circularly polarized light". Circular polarizing filters are actually sandwiches of (a) a regular polarizing filter and (b) a piece of plastic which causes a retardation of 1/4 wave (typically about 140nm).
Why is all this important? And what does it have to do with image analysis? The biggest challenge in image analysis is simplifying objects so that they can be recognized by an automated measuring system. In polarized light, birefringent objects viewed between crossed polars are bright against a dark background ... great for segmenting with an automated system. Unfortunately, the intensity changes with orientation (yet another discussion). The same crystal, for example, will appear dark if sitting N-S or E-W but bright at +/- 45 degrees! On the other hand, if you use crossed CIRCULAR polars (make sure the retarding plastic is toward the center of the "sandwich" and the sample is in between), all the orientation effects will be removed because of the corkscrew path of the light. The result: all birefringent objects which are behaving the same would appear the same intensity, regardless of orientation.
We used this trick very successfully, for example, with the Center for Urban Ecology, when they were trying to measure the % crystallinity of soil samples.
Note: if you are interested in trying circularly polarized light, you can ask your microscope rep to lend you a set of circular Pol filters or you can make your own, using 2" squares of polarizing filter and "quarter wave" material, typically available from companies like Edmund Scientific. (Caveat: no financial interest).
Whew! A long winded answer, but I hope that it is helpful.
If you are interested in copies of the diagrams described above, contact me off line.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
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Thanks to all who responded to the recent inquiry about forensic applications. Once again this is a wonderful site because of the people willing to share their knowledge.
There's a fingerprint exercise in Project MICRO's "Microscopic Explorations", which was written for grades 4-8. "Microscopic Explorations" is published in the Lawrence Hall of Science GEMS series, which also includes "Mystery Festival", (crime scene investigation) and "Fingerprints"; you'll find a link to GEMS on the MICRO website (URL below). The Minnesota Microscopy Society has an excellent outreach website, http://resolution.umn.edu/MMS/ProjectMicro/ : you'll find a link to fingerprints at NIST and images of several fabric types there. Don't miss Joe Neilly's delightful "Beanie Baby Mystery" in the Classroom Activities part of the MICRO site. Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Dear Ms. Walsh, This is not a reply to the general list but a remark that I hope will be taken as reflective. I know that interesting "hooks" are needed to get children interested in science with all the ramifications that a scientific grounding has for our future, but isn't the constant diet of mayhem, down to the most grotesque details of forensic science, being served up from enough sources already? I think that the constant stream of "forensic" programs and cop shows conditions children to an acceptance of a police state mentality. The same instructional subjects can be approached from the biological and earth sciences. (Microscopy to solve the mysteries of developing life; micro-fossils in common stones that explain the mystery of what happened on earth when there was no one yet to observe it, etc.) If you want to deal with fibers, how about the (relatively) benign world of counterfeit goods - the alpaca cape that turns out to be rayon, the shawl that turns out to be shanoosh from poaching of endangered antelope. In fact the whole area of wildlife forensics, the evidence gathered in the protection of endangered species, at least avoids more of the human-on-human violence that is made to seem the norm by its familiarity. (See the website of the federal wildlife forensics lab in Ashland, Oregon.) John Twilley
Rosemary, www.mccrone.com will lead you to some interesting things and Ted Clarke's pages have some information about asbestos http://www.couger.com/microscope/Ted-Clarke/ Good luck Gordon Gordon Couger gcouger-at-couger.com
Rosemary You should look up GSR (Gun Shot Residue Analysis), this is commonly used with a SEM and EDS. Try this URL: http://www.gunshotresidue.com/ JQ Jim Quinn {jquinn-at-/" EUDORA="AUTOURL"www.matscieng.sunysb.edu}
} From: "Ann St. Amand" {astamand-at-phycotech.com}
Jon, What you want to do is a leaf dip. Use a formvar + cabon grid held in a tweezer. Put a drop of uranyl acetate on the grid. Then take a piece of what appears to be an infected leaf, macerate an edge with a razor blade and drag it through the stain droplet a few times. This will deposit some of the sap containing some organelles and virus, if present, onto the grid. Wick off the remaining stain and view. TMV is very easily recognized as a rod shaped virus. You might find that the problem is another virus type so also look for round viruses or viruses that are rod shaped but of very unequal length. TMV is about 18nm wide. I forget exactly how long it is but probably not more than about 200nm. Particles are quite regular in length.
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
On 9/15/03 1:50 PM, "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} 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: } } Is there a quick and dirty method for looking at TMV? } } I have never done it, a person from the local community wants to see if } that's what killing his plants and insists on 'seeing' it with his own } eyes. } } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
I've used RR to look at the extracellular matrix in cardiac muscle and to label the apical surface of cultured retinal pigment epithelial cells. You do not need to use it after the osmium step. don't be surprised if the labelling is patchy. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
General information: http://www.shortcourses.com/how/microscope/microscope.htm http://www.microscopy-uk.org.uk/mag/artaug01/vrcoolpix.html
Another way is to build the adapter yourself. I did that myself with my Canon G3 and our old Leica microscope. It is equipped with an old analog camera, and the only thing I had to build was a small tubus which fits to the old eyepiece of the analog camera (and holds the camera in the right position above it). Leica sells a similar system for app. 2000 €... (Adapter and camera).
I have to admit that the Canon G3 is not the best camera for microscopy. It is a fine camera for normal pictures, and I really like it. "But" it's lense is quite large in diameter. That is nice, because it get's a lot of light, but the openings of eyepieces I know of are not large. Therefore, if you don't zoom in digitally (4x optical zoom is not enough), you always have circular black frames on the picture. The smaller the lens of the camera, the better for microscopy (I think). (But I bought the camera, b/c I can easily use a ring flash with it). Also, the part of the field in view is quite small. You have to take more then one picture to get all information you see through the eyepiece.
Another topic is the software you want to use to aquire pictures with. It is not nice to handle the camera itself when attached to the microscope (even if it has a display you can flip to the right direction).
All Canon cameras come with a software called "remote capture". With it, most functions of the camera can be controlled via USB from the computer. (The large hard disk is quite nice for time lapse recordings... Having space for } 10000 pictures (at 4 MP) is definitly different to a 36 picture film.) Another software from the original package (well, designed for preparing a panorama photo out of a number of single pictures) is quite nice to put single pictures together in a matrix - we just tried it out with 40 single C. elegans pictures - now we have a worm picture with app. 100 MP... Quite large and difficult to handle... And who want's to print that out??
:-) Torsten
Torsten Fregin
Universität Hamburg - Zoologisches Institut Abt. Neurophysiologie AG Wiese - Raum 413 Martin-Luther-King-Platz 3 20146 Hamburg, Germany Telefon *49-(0)40-42838-3931 Fax *49-(0)40-42838-3937 eMail Torsten.Fregin-at-zoologie.uni-hamburg.de Torsten-at-Fregin.de
The C2 apertures in our CM200 FEG has become very dirty and needs to be replaced as soon as possible. As you might be aware, the C2 apertures for FEG have to be very clear and need to take a special method for cleaning. Now I have got several options:
A.Buy from FEI directly but they obviously cost about more than three times from ordinary aperture of which if you purchase from Agar. B.Use special plasma cleaner to clear up those ordinary apertures so that apertures would be acceptable by FEG. C.Try to use the plasma from a carbon coater to clear up ordinary apertures.
My questions is either B or C would work? Alternatively, there are any good ideas for this purpose?
Many thanks!
Peiyi Manager of EM Krebs Institute for Biomolecular Research University of Sheffield Firth Court Western Bank Sheffield Yorkshire S10 2TN United Kingdom Tel: +44 (0)114 222 2000 Direct: +44 (0)114 222 2739 FAX: +44 (0)114 272 8697 E-mail: p.wang-at-sheffield.ac.uk
I have an additional remark. Sometimes it is useful to take the measured background (Bremsstrahlung) as reference to the beam current. That works only, if the unknown specimen and the standard have nearly the same mean atomic number. If not, the Bremsstrahlung depends from Z and this fact has to taken into account. If analyst takes the Bremsstrahlung reference, a very high precission is reachable, because charakteristic radiation and Bremsstrahlung intensities are both directly depend from beam current. It is useful, if a standard exists with nearly the same elements and concentrations as the expected in unknown.
Otherwise: If the beam current vary in percent units, the results must vary in same order of magnitude (in standard comparison analysis), too. Only normalization can help, like Warren remarked. But this works only if all elements are possible to measure and were measured in same time. That is why standard comparison analysis often do not have better results than standardless. Finally, to overcome the problem with normalization in standardless analysis, you need the Bremsstrahlung background, too.
Frank Eggert
By the way, for all who have got a Palm Handheld and need line energies (keV) or a periodic chart of elements for your Palm } } } try: http://www.microanalyst.net/software.phtml
______________________________________________________________________________ Zwei Mal Platz 1 mit dem jeweils besten Testergebnis! WEB.DE FreeMail und WEB.DE Club bei Stiftung Warentest! http://f.web.de/?mc=021183
You may want to take a look at our webpage on connecting Coolpix cameras to microscopes: http://www.mvia.com/Coolpix/clpxadpt.htm
There is a lot of information there about different types of microscopes as well as various options for how to mount the camera onto your microscope.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
"max_gra-at-libero.it" 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´d like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. } I´d like to have a high resolution. } Could someone give me a suggestion about the best device (as brand) to apply? } Thank you. } Best Regards, } Massimo
I got plenty of advice about how to do a quick survey for TMV. Most of it was what I had suspected, a simple leaf drag through some negative stain. Though I had never done it before, it was reassuring to know I was on the right track.
The guy came to the lab today, we did not see any TMV in his leaves. The leaves looked mottled, like TMV, but I did not see any in the EM. I tried 3 grids, got lots of other junk on the grids, but either no TMV or there was so much of it that it made a solid mat that made seeing individual particles impossible.
Now, here's the good part. He came in with a couple of species of tobacco leaves, they looked like they had TMV. I said "Looks like TMV, nothing you can do about it, doesn't matter if we can see virus or not, the plants are toast."
He said he really wanted to know if it was TMV because he had other plants growing and was afraid they might get infected too. As we talked over other ways of confirming what was wrong with his plants, he started to tell me the whole story.
Seems he is growing these plants for 'ceremonial' purposes. Got the seeds from a shamans garden in New Mexico and one of the species he had contains up to 15% nicotine, he says. He was concerned that he might have to 'sacrifice' the rest of the plants if this was TMV, he was hoping there might be some alternative to starting over.
Only in Santa Cruz ................
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
The leaf dip technique is valuable for locating plant viruses but not always 100% trustworthy, even when plenty of virus is present. Back in the 1960s a colleague of mine was studying plant viruses and used PTA as the negative stain. He saw plenty of rods (it was a rod-virus, but not TMV), however nothing else. For some now unknown reason he did another leaf dip but this time with a different negative stain, (uranyl acetate or ammonium molybdate?), and saw lots of small icosahedral virus particles but no rods. After much speculation about the reason for these "spherical rods" he concluded that this was a mixed (double) viral infection. This was later proved correct. The lesson here is not to rely solely on one negative stain when examining an unknown infection, especially when no virus can be seen. It could be that the stain is for some reason not capable of contrasting the virus; try another stain to be sure. Best wishes,
Jim
-----Original Message----- } From: Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Wednesday, September 17, 2003 12:00 AM To: Microscopy-at-sparc5.microscopy.com
Hi:
I got plenty of advice about how to do a quick survey for TMV. Most of it was what I had suspected, a simple leaf drag through some negative stain. Though I had never done it before, it was reassuring to know I was on the right track.
The guy came to the lab today, we did not see any TMV in his leaves. The leaves looked mottled, like TMV, but I did not see any in the EM. I tried 3 grids, got lots of other junk on the grids, but either no TMV or there was so much of it that it made a solid mat that made seeing individual particles impossible.
Now, here's the good part. He came in with a couple of species of tobacco leaves, they looked like they had TMV. I said "Looks like TMV, nothing you can do about it, doesn't matter if we can see virus or not, the plants are toast."
He said he really wanted to know if it was TMV because he had other plants growing and was afraid they might get infected too. As we talked over other ways of confirming what was wrong with his plants, he started to tell me the whole story.
Seems he is growing these plants for 'ceremonial' purposes. Got the seeds from a shamans garden in New Mexico and one of the species he had contains up to 15% nicotine, he says. He was concerned that he might have to 'sacrifice' the rest of the plants if this was TMV, he was hoping there might be some alternative to starting over.
Only in Santa Cruz ................
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
We are in the process of purchasing a 4K x 4k or 2k x 2k CCD camera for our JEOL 2010F TEM. The microscope is attached with Gatan's Digipeels, therefore the CCD camera has to be compatible with the existing Peels system. Our questions are:
1) Is Gatan the only provider for such kind of CCD camera? If there is other providers, we would like to consider.
2) Is the technology for the 4k x 4k camera still premature?
Any other input regarding CCD camera are welcome.
Thanks a lot!
James
_________________________________________________________________ Compare Cable, DSL or Satellite plans: As low as $29.95. https://broadband.msn.com
We are in the process of purchasing a 4K x 4k or 2k x 2k CCD camera for our JEOL 2010F TEM. The microscope is attached with Gatan's Digipeels, therefore the CCD camera has to be compatible with the existing Peels system. Our questions are:
1) Is Gatan the only provider for such kind of CCD camera? If there is other providers, we would like to consider.
2) Is the technology for the 4k x 4k camera still premature?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jjjani-at-gauanand.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, September 16, 2003 at 22:43:21 ---------------------------------------------------------------------------
Email: jjjani-at-gauanand.com Name: Janardan Jani
Organization: Gujarat Agricultural University
Education: Graduate College
Location: Anand, Gujarat State, India
Question: Respected Sir/ Madam,
I wish to carry out SEM for my Bacterial isolates which are probably Bacillus thuringiensis. I wish to observe the spore and crystal the delta endotoxin which is bipyramidal in shape. I have access to Hitachi S3000 SEM but do not have protocol for the sample preparation. I would be obliged if i am given a protocol about the sample preparation and staining if any for the purpose.
I keep my ethanol series (50, 70, 95, and 100%) in squirt bottles that have a long tube like shaft coming out the top of them. If you pull the shaft upwards, it seals off the bottle and prevents evaporation and minimizes humidity contamination of the ethanol. Unfortunately, these bottles are getting quite old (16 years) and need replacement. I can't remember where I got them. Does anyone use a similar type or have a better alternative? Thanks, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Another CCD camera company worth considering is TVIPS. They are in Germany and they also have representatives in the USA. They make very good cameras and easy to install and use. The contact details are:
Hans Tietz Email: hans.tietz-at-tvips.com Fax: +(49)-89-8509488
Sincerely,
Thearith Ung
-----Original Message----- } From: Boron nitride [mailto:boronnitride-at-hotmail.com] Sent: Wednesday, September 17, 2003 5:21 AM To: Microscopy-at-sparc5.microscopy.com
Dear colleage,
We are in the process of purchasing a 4K x 4k or 2k x 2k CCD camera for our JEOL 2010F TEM. The microscope is attached with Gatan's Digipeels, therefore
the CCD camera has to be compatible with the existing Peels system. Our questions are:
1) Is Gatan the only provider for such kind of CCD camera? If there is other
providers, we would like to consider.
2) Is the technology for the 4k x 4k camera still premature?
Any other input regarding CCD camera are welcome.
Thanks a lot!
James
_________________________________________________________________ Compare Cable, DSL or Satellite plans: As low as $29.95. https://broadband.msn.com
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Peiyi Wang wrote the following: ================================================================= } The C2 apertures in our CM200 FEG has become very dirty and needs to be } replaced as soon as possible. As you might be aware, the C2 apertures for FEG } have to be very clear and need to take a special method for cleaning. Now I } have got several options: } } A.Buy from FEI directly but they obviously cost about more } than three times from ordinary aperture of which if you } purchase from Agar. } B.Use special plasma cleaner to clear up those ordinary } apertures so that apertures would be acceptable by FEG. } C.Try to use the plasma from a carbon coater to clear up } ordinary apertures. } } My questions is either B or C would work? Alternatively, there are any good } ideas for this purpose? =================================================================== You always have the option of purchasing new apertures from either the original equipment manufacturer or from other sources, such as Agar (as you indicated) or SPI Supplies or yet others.
A plasma cleaner operates at very low power (supposedly under 10 watts) whereas a plasma etcher operates typically at 100 watts or higher. The quality of the cleaning will depend on the power being used, and since there is no reason not to use 100 watts, you could clean the apertures in a few minutes or less in a plasma etcher using oxygen in a unit such as the SPI Plasma Prep™ II plasma etcher. See URL www.2spi.com/catalog/instruments/etchers1.shtml
If you were using a plasma cleaner, such as the SPI Plasma Cleaner, see URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml you could use a mixture of oxygen and argon, which would be more effective. You don't want to use argon at 100 watts since it could possibly etch the aperture itself, which obviously would be undesireable.
If you use this approach, your apertures will last nearly forever (well nothing is forever) since you are not heat cycling them as you would in a vacuum evaporator. Of course, users eventually (accidentally) damage the edges of the hole with the beam so apertures really don't last forever. But cleaning them this way will enable them to last far longer than would otherwise be possible.
We don't see how you could use the plasma in a carbon coater to clean apertures, we have never heard of anyone doing that. Are you sure you are not referring to the use of a vacuum evaporator (with diffusion or turbo pump) to clean apertures, but in that case, it would be the heat instead of a plasma responsible for the cleaning.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHmR5t013948 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 12:48:27 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HHmRtO013944 for L1; Wed, 17 Sep 2003 12:48:27 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-sparc5.microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HHle5t013914 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 12:48:06 -0500 Received: from nightshade.noc.ucla.edu (nightshade.noc.ucla.edu [169.232.48.18]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHl65t013904 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 12:47:23 -0500 Received: from azalea.noc.ucla.edu (azalea.noc.ucla.edu [169.232.47.12]) by nightshade.noc.ucla.edu (8.12.8p1/8.12.8) with ESMTP id h8HHkqW9021247 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 10:46:52 -0700 Received: from emi.ucla.edu ([149.142.241.180]) (authenticated bits=0) by azalea.noc.ucla.edu (8.12.9/8.12.9) with ESMTP id h8HHkp7T007203 (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NOT) for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 10:46:51 -0700 Message-Id: {5.2.0.9.2.20030917102256.01b30cf8-at-mail.ucla.edu} X-Sender: sryazant-at-pop.bol.ucla.edu X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9
Dear James Gatan is not only producer of high resolution digital cameras for TEM. You may shop around to find other manufacturers (or they will find you). Personally, I do have camera from Gatan and really happy with it. A few things to consider:
- CCD cameras with fiber-optics coupling (most modern cameras including Gatan's UltraScan 4x4 or 2x2) are slow. It means, you don't have real video-mode. It does not matter what manufacturer claims, in reality you do have a few frames per second and that's it. Again, as more pixels you have, camera is slower (in general). To overcome this problem, Gatan virtually split their 4x4 chip on 4 quadrants and read them simultaneously increasing speed 4 times. You have to pay a fortune for such trick.
- as far as I know, UltraScan 4x4K is pretty mature camera; they install them on new FEI TEMs as a part of standard package. If I will have money, I'll definetely get it (camera and FEI microscope as well).
- Another things to consider is customer service.
- Before you made final decision, ask company for the demo. When I did shopping for my camera, I had a few offers from different companies. The cameras were pretty similar on the paper and so different when I took pictures and compare them side-by-side. So, ask for the demo, bring your own samples, took your own pictures and compare.
Good luck, Sergey.
At 05:37 AM 9/17/2003, 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 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bart-at-lasalle.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 17, 2003 at 09:52:23 ---------------------------------------------------------------------------
Email: bart-at-lasalle.edu Name: Henry Bart
Organization: La Salle University
Title-Subject: MListserver:
Question: I have a TOPCOM SM-300 SEM and would like to know what company,if any, has picked up that line after RJ LEE stopped supporting it. I need some parts and advice. Thanks
I agree with Lee's answer, RR in the glut, rinses, osmium and after that no RR. I also used the Luft protocol. I seem to remember using para-phenylene diamine (0.5% in alcohol during dehydration) to intensify (reduce) the osmium-RR complex on at least some of my samples. I had fantastic contrast, almost too much. Since I was looking at cells in culture evenness of penetration/staining was not a problem. By some strange coincidence I also used RR on cultured retinal pigment epithelium just as Lee did.
Geoff
Tom Phillips wrote:
} I am using Luft's technique for staining the glycocalyx with Ruthenium } Red. I know the RR is supposed to be included in the aldehyde fix, } the rinses and the osmium but how about the osmium rinses or ethanol } dehydration series? Has anyone successfully done this procedure? } Thanks, tom } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
-- -- ********************************************** 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 **********************************************
We need a hint or two about cryosectioning nylon. The cryocutter in our lab is used to sectioning complex hydrated heterocopolymers (i.e. biological specimens), and the nylon is being recalcitrant. Any tips (I checked the U. Florida Tips & Tricks site) for cutting temperature, cutting speed and so forth? This is for routine TEM study of a 2-phase nylon, and a nylon with silicon dioxide nanoparticles. Nothing extraordinary. Thanks for any hints.
Phil
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Tom, As a substitute for your ethanol bottles I would suggest that you consider the pull-up type caps that are available on some juice and water bottles. I have noticed that there is a wide variety of sizes and quality to the different suppliers. Perhaps there is one that fits glass bottles or better plastic ones, like the pint bottles that the ethanol comes in.
I hadn't thought of using these before but it seems like a good idea to try.
I have not been able to get the latest version (Beta 4.0.2) of Scion Image (on a machine running Win XP) to open or import digital images. I've tried to open and import both .tifs and .bmps to no avail. The program just crashes. Scion does not provide technical support unless you have purchased equipment from them, which I have not. Can anybody suggest a solution? Or, alternatively, do you know of any other image analysis software that's free/inexpensive? Thanks!
ImageJ put out by NIS is free and works on almost anything. http://rsb.info.nih.gov/ij/ It is the successor of Sicon image.
Gordon } From: "Diana Greenlee" {greenlee-at-u.washington.edu}
: : I have not been able to get the latest version (Beta 4.0.2) of Scion Image : (on a machine running Win XP) to open or import digital images. I've : tried to open and import both .tifs and .bmps to no avail. The program : just crashes. Scion does not provide technical support unless you have : purchased equipment from them, which I have not. Can anybody suggest a : solution? Or, alternatively, do you know of any other image analysis : software that's free/inexpensive? Thanks! :
I need to purchase a frame grabber card for Image Analysis to use either NIH Images Analysis, Scion program or the Image Java Program but I need to know what boards will work with an Amray 1000 SEM. I have both a MAC G3 and PC Windows platforms.
The short answer is I don't know. But I would ask you first, are you sure a frame grabber will provide you enough resolution?
I have nothing against Scion or any other manufacturer of frame grabbers, but I am not sure that you really want a frame grabber for SEM applications. The limitations of the video signal, RS-170, prevent you from digitizing more than about 640 pixels across the image, and that assumes that the Amray uses RS-170. A frame grabber may be adequate for basic imaging, but the image will be somewhat grainy. I would not be inclined to publish such images or to attempt image analysis on them.
I think most SEM imaging systems resort to either active or passive digitization to collect images at higher resolution.
Warren
At 06:32 AM 9/18/2003 -0700, you wrote:
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Lookingfor a method of determining live vs dead via microscopy BUT not using a fluorescent stain. Organism is a level 2 pathogen and cannot be accomodated safely under our fluorescent scopes.
The samples do not need to be live at time of observation.
If there is a live dead fluorescent stain which retains live dead after fixation/killing that would work as well.
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 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
Years ago, when working on viability of ovine articular cartilage, I noticed that fresh tissue stained with acridine orange maintained its viability status (as judged by colors of the dye) after it had been kept in 70% ethanol at 4oC for several days.
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Thursday, September 18, 2003 10:51 AM To: microscopy-at-MSA.Microscopy.com
Lookingfor a method of determining live vs dead via microscopy BUT not using a fluorescent stain. Organism is a level 2 pathogen and cannot be accomodated safely under our fluorescent scopes.
The samples do not need to be live at time of observation.
If there is a live dead fluorescent stain which retains live dead after fixation/killing that would work as well.
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 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
Zeiss TEM 10A has to be moved ASAP. It was in use and working well in July 2002. It was serviced in June 2002. Contact Dr. Cartun at the following address: Rcartun-at-harthosp.org It is located in Connecticut.
I wonder if it is possible to use 6 nm-gold-tagged antibodies for pre-embedding immunolabeling, or if it is necessary to use smaller ones? I want to do immunolabeling of betacells from the islets of Langerhans (rat).
Sincerely, Pernilla Nevsten
........................................................................................................................................... Pernilla Nevsten, PhD-student Materials Chemistry Lund University, Sweden
Question: I¥we used Ruthenium Red to stain the capsule of vibro colerae successfully. I use RR in the fixation, rinse and osmium but not in the osmium rinse or the ethanol dehydration steps. Good luck!
We are seeking information from anyone who has knowledge of how long a critical-point dried specimen prepared for SEM is sufficiently stable for subsequent viewing. We anticipate receiving many samples of glutaraldehyde-fixed plankton with delicate protozoa and algae in them. Our colleagues sending the material will eventually sample only a small subset based on data they gather one year from now. So, we will be asked to examine the stubs approximately 12 months in the future.
Has anyone advice on the stability of specimens for SEM observation that have been placed on stubs one year prior to observation? Is it preferable to lightly sputter-coat the sample before storage and is there any secure hope of stabilizing delicate flagellar structures, etc. after one year? We understand the importance of storage in a dessicator and at fairly constant temperature, but we have never stored specimens for 1 year before observing them.
Many thanks, Dee
```````
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
I would like some opinions about critical point dryers. What do you think about automated units vs. manual units. Or do you use HMDS and not use the CPD at all? If you could post to the list and not send them directly to me, I would appreciate it and everyone could read the responses.
Thanks, Jean Ross Central Microscopy Research Facility University of Iowa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pfraundorf-at-umsl.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, September 20, 2003 at 08:45:53 ---------------------------------------------------------------------------
Email: pfraundorf-at-umsl.edu Name: Phil Fraundorf
Organization: UM-StL Physics and Astronomy/CME
Title-Subject: Education: a variable size-scale adventure
Question: We've been looking for web strategies to let non-microscopists share the experience of moving between macroscopic and nanoscopic size scales for some time. The link below* is a step in that direction. The puzzlers beneath the model are meant to be experimental in nature, rather than based on theory, so put on your Sherlock Holmes hat and enjoy.
Also, what are some: (i) learning challenges and storylines for use by students and teachers with an exploration tool like this; and (ii) ideas for scenarios to that end, e.g. different specimen configurations? The first model you encounter uses a fixed intersecting-axis goniometer. The beta model allows movement of that rotation point with respect to a "scope-based" cartesian system by hitting x,X,y,Y,z,Z. We also have a way to request lateral displacements accompanied by a dynamically redrawn heirarchy of grids. Although this can't yet be done at display rates (i.e. many times per second), current tools do allow us to generate "data" (e.g. simulated HREM images, diffraction patterns, X-ray or roughness spectra, etc.) from regions of interest. We're also interested in feedback from storyline field tests in university and high school classrooms, as well as in pooling resources for this between institutions.
Phil Fraundorf UM-StL Physics & Astronomy/CME Washington U. Physics/MCSS office: (314)516-5044 lab: (314)516-5024 pfraundorf-at-umsl.edu http://www.umsl.edu/~fraundor
The Midwest Microscopy & Microanalysis Society would like to announce two upcoming meetings this fall. Please (!) circulate this notice to all your colleagues.
Friday, October 3rd University of Wisconsin, Room 235 USDA Dairy Forage Research Laboratory, 1925 Linden Drive West, Madison, Wisconsin 53706. 9:00AM to 4:30PM See the below link for the complete program: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSOct2003.html
Changes to the posted program are as follows:
Ranier Guillery, Professor Emeritus UW Department of Anatomy, UW Madison will replace David Slautterback
In the afternoon Biological session: Sara Patterson's title is: "A look at cell separation in Arabidopsis using floral organ abscission as a model system"
In the afternoon Physical Sciences Session (moved to 3:05pm) : Mark Eriksson' s title is : " Novel scanning probe techniques: from off -angle tapping to dielectric imaging".
Tom Kelly from Imago Scientific will open this session (at 1:pm) with a talk entitled; "3D Atomic- Scale Compositional imaging with local electrode Atom Probes.
Friday, November 14th Harper College, Wojcik Conference Center, 1200 West Algonquin Road, W108, Palatine, Illinois 60067 SEM Workshop : Stuart McKernan - University of Minnesota: Environmental SEM John Mackenzie - North Carolina State University: Digital Imaging Robert Apkarian - Emory University: Cryo SEM in Biomedical Sciences Wayne Neimeyer- McCrone Associates- Forensic Microscopy
Details and www link to follow.
For further information please contact either:
Robert Mierzwa (MMMS President - Elect): Tel: 920-803-8945, email: mierzwa-at-jeol.com Arvid Casler (MMMS Program Coodinator): Tel: 847-674-7700, email: arvid_casler-at-fmo.com Jim DiOrio (MMMS President): Tel: 847-270-4676, email: jim_diorio-at-baxter.com
Dear Pernilla, In my opinion, you can use 6 nm gold tagged antibodies only if your cell is capable of endocytosis. Otherwise, you have to permeabilize your cell and use ultra small gold coupled to your antibody. Another alternative is to do a common immunolabelling, after permeabilization ( antibody of interest, then species specific ultra small gold.) In all case you have to amplify your small gold particles to visualize them in a electron microscope. Best regards danièle -------------------------------------------- Danièle Spehner, INSERM EPI 99-08 - EFS-Alsace 10 rue Spielmann - 67065 STRASBOURG tel : 03 88 21 25 25 - fax : 03 88 21 25 44 e-mail : daniele.spehner-at-efs-alsace.fr ------------------------------------
-----Message d'origine----- De : Pernilla Nevsten [mailto:pernilla.nevsten-at-materialkemi.lth.se] Envoyé : lundi 22 septembre 2003 08:16 À : Microscopy-at-MSA.Microscopy.Com Objet : pre-embedding immunolabeling
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Dear Microscopy Listserver members,
I wonder if it is possible to use 6 nm-gold-tagged antibodies for pre-embedding immunolabeling, or if it is necessary to use smaller ones? I want to do immunolabeling of betacells from the islets of Langerhans (rat).
Sincerely, Pernilla Nevsten
.......................................................................................................................................... Pernilla Nevsten, PhD-student Materials Chemistry Lund University, Sweden
As long as the samples are properly stored with good desiccant, they are archival. A year is no problem. I would suggest not sputter coating them, since you know you won't be looking at them for a year. Protection against mechanical shock -- jarring and the like -- is the best way to preserve delicate structures. But. Do you or your colleagues already know the proper and best methods for preparing the critters? Some might require OsO4, for example, and what's best for ciliates isn't necessarily best for dinoflagellates and so on. Meaning, it might be a good idea to process and examine some samples of each kind of wee beastie now, rather than waiting, to make sure they're actually processed right.
Phil
} Dear colleagues, } } We are seeking information from anyone who has knowledge of how long } a critical-point dried specimen prepared for SEM is sufficiently } stable for subsequent viewing. We anticipate receiving many samples } of glutaraldehyde-fixed plankton with delicate protozoa and algae in } them. Our colleagues sending the material will eventually sample } only a small subset based on data they gather one year from now. So, } we will be asked to examine the stubs approximately 12 months in the } future. } } Has anyone advice on the stability of specimens for SEM } observation that have been placed on stubs one year prior to } observation? Is it preferable to lightly sputter-coat the sample } before storage and is there any secure hope of stabilizing delicate } flagellar structures, etc. after one year? We understand the } importance of storage in a dessicator and at fairly constant } temperature, but we have never stored specimens for 1 year before } observing them. } } Many thanks, } Dee } *************************************************************** } Please do not publicly post any of my correspondence without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995)
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Manual, not automatic. The automatic CPDs I've seen are too inflexible, and I don't trust them anyway. Maybe I'm a Luddite, but I don't even like "semi-automatic" CPDs. And yes, I do use HMDS, but it's not appropriate for everything.
Phil
} Hi everyone, } } I would like some opinions about critical point dryers. What do you think } about automated units vs. manual units. Or do you use HMDS and not } use the CPD } at all? If you could post to the list and not send them directly to me, I } would appreciate it and everyone could read the responses. } } Thanks, } Jean Ross } Central Microscopy Research Facility } University of Iowa
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hi all, I reply as an old foggie probably to set in my ways,but here goes. Over the past 30 or so years I have used several CPD's usually with acceptable results, however the machines I have had most unpredictable results with were those with automatic cycling and programmed temperture rise. The simple Polaron and others with manual valves and tap water temperature controlled by a shower mixing control have been most reliable and trouble free.
Good Luck,
Ron.
jean-ross-at-uiowa.edu 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 everyone, } } I would like some opinions about critical point dryers. What do you think } about automated units vs. manual units. Or do you use HMDS and not use the CPD } at all? If you could post to the list and not send them directly to me, I } would appreciate it and everyone could read the responses. } } Thanks, } Jean Ross } Central Microscopy Research Facility } University of Iowa
-- Ronald J. Smith Department of Biology Room 235, Biological & Geological Sciences Bldg. U.W.O., London, Ontario N6A 5B7 (519) 661-2111 ext.86486
Dee I have re-examined CPD virus specimens after 9 months to a year and they looked pretty much as they did at first. I think you have every chance of success, particularly if you store the specimens in a desiccator at a steady temperature as you suggest, and also preferably in the dark. I would recommend not coating them until you are going to examine them. Chris
University of Edinburgh Biological Sciences EM Facility
----- Original Message ----- } From: "Dee Breger" {micro-at-ldeo.columbia.edu} To: {microscopy-at-msa.microscopy.com} Sent: Monday, September 22, 2003 3:14 PM
For the last several years, we have been coating our glass microscope slides with aminopropyltriethoxysilane. We have found it superior to gelatin, chrome-gel, poly-D-lysine, etc for increasing adherence of both cryostat and plastic resin sections. We love this technique but after years of success, it has stopped working or at least fully working. One problem is decreased adhesion and lost sections during rigorous immunostaining protocols. Another easily seen difference is that water droplets no longer stay beaded up due to surface tension but spread over the surface more easily. We have tried two new bottles (different sources) and new acetone bottles. We tried doubling the APTS concentration to 4% but it made little difference. Does anyone have insights into this problem? I am not really looking for an alternative coating material but would rather solve the problem with this one. Our actual protocol is: Dip slides in 2% aminopropyltriethoxysilane in acetone for 1 min followed by 1 min in 100% acetone, then 1 min in dH20, allow to air dry. Thanks in advance for your comments. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I was discarding old files today when I noticed a paper from the old (undated) EMSA Bulletin by Christine Jeffery of Sothern Illinois University. She found that some osmium treated, non-coated SEM samples stored in one vendor's sample box with a soft plastic insert developed crystals after about six months. Metal coated samples did not exhibit the crystal formation. The implication was that an unknown mechanism involving osmium laden samples and a phenol compound in the box inserts resulted in the crystals. So perhaps it is wise to metal coat SEM samples if they will be retained for study after six or more months. Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140, Room S101 San Francisco, CA 94143
Your submission of other meetings will be very appreciated (submission form is accessible from the page mentioned, the direct link is http://www.petr.isibrno.cz/microscopy/PMRform.php).
Petr Schauer +--------------------------------------------------------------------+ | Dr. Petr Schauer tel.: +420 541 514 313 | | Head of the Group of the Scintillation fax : +420 541 514 404 | | and Cathodoluminescent Systems +420 541 514 402 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | | Czech Republic www: http://www.petr.isibrno.cz/ | +--------------------------------------------------------------------+
Hi, Is Gatan the only commercial vendor offering CL on SEM's? Have any of you heard of a company called KNE?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
I wonder if it is possible to use 6 nm-gold-tagged antibodies for pre-embedding immunolabeling, or if it is necessary to use smaller ones? I want to do immunolabeling of betacells from the islets of Langerhans (rat).
Sincerely, Pernilla Nevsten
........................................................................... .............................................................. Pernilla Nevsten, PhD-student Materials Chemistry Lund University, Sweden
I usually use something along the line of the Nanoprobes 1.4nm nanogold for pre-embedding labeling, the cells are permeabilized after primary fixation. You can enhance it after the labeling step (before embedding) to make it easier to locate in the microscope.
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
I believe Gatan may be the only commercial vendor of a well developed CL product with a presence in the U.S., however: There was an article/short paper in the May 2003, issue 60 (The Americas edition) Microscopy and Analysis that used a CL system from Japan (on a JEOL 6500F thermal FE-SEM) (the study was conducted at Kyoto Inst. of Tech., Japan) The CL device was a MP-32FE, Atago Bussan, Horiba Group, Tokyo The capabilities and features available as described in the paper seem similar to Gatan offerings, but I never investigated further than reading the paper. (We already own a Gatan MonoCL3 system)
Probing Nanoscopic Stresses in Glass using Luminescent Atoms Giuseppe Pezzotti, Ceramic Physics Laboratory & Research Inst. for Nanoscience, Kyoto Inst. of Tech., Japan
If you'd like the author's contact info, a copy of the paper, or have questions about our Gatan system, feel free to contact me.
------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
This reminds me I was looking for the next Microscopical Society of Canada meeting the other day. If anyone reading this is associated with that meeting, then please make note of the MSC wwwpage is way out of date. http://msc.rsvs.ulaval.ca/english/welcome.html ... and I didn't get a response from the executive secretary because the e-mail address is no good.
What I have been able to find, is that it is 'apparently' being held at Acadia University, Nova Scotia ... but consider it heresay.
} Dear Microscopists, } } An update of the list of meetings for microscopists in the year 2004 } has been finished at the Petr's Microscopy Resources at the } } http://www.petr.isibrno.cz/microscopy/meetings.php#2004 } } Your submission of other meetings will be very appreciated (submission } form is accessible from the page mentioned, the direct link is } http://www.petr.isibrno.cz/microscopy/PMRform.php). } } Petr Schauer } +--------------------------------------------------------------------+ } | Dr. Petr Schauer tel.: +420 541 514 313 | } | Head of the Group of the Scintillation fax : +420 541 514 404 | } | and Cathodoluminescent Systems +420 541 514 402 | } | INSTITUTE OF SCIENTIFIC INSTRUMENTS | } | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | } | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | } | Czech Republic www: http://www.petr.isibrno.cz/ | } +--------------------------------------------------------------------+ } } } }
I've just realized that the current frames format of the MICRO web page requires that you access all of its segments (the bibliography, for example) via the URL below. If you have a different bookmark stored, please change. -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I think that Mike is referring to KE Developments in the UK. Their website with information regarding their detectors (including CL) is at http://www.kedev.co.uk/detectors.htm.
best regards, Steven Slap
on 9/23/03 8:17 AM, Michael Cheatham at mmcheath-at-mailbox.syr.edu 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, } Is Gatan the only commercial vendor offering CL on SEM's? } Have any of you heard of a company called KNE? } } TIA } Mike
Your reference to KNE, may be KE Developments in the UK. Energy Beam Sciences has recently become the US distributor for KE Developments. You may check the KE Developments web-site www.kedev.com, contact me at 800-992-9037 X340, or by e-mail. for assistance on CL detectors
Regards,
Mike Dufraine Energy Beam Sciences, Inc.
-----Original Message----- } From: Michael Cheatham [mailto:mmcheath-at-mailbox.syr.edu] Sent: Tuesday, September 23, 2003 8:18 AM To: Microscopy-at-sparc5.microscopy.com
Hi, Is Gatan the only commercial vendor offering CL on SEM's? Have any of you heard of a company called KNE?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
I believe AOL & Compuserve & Netscape.Net Email users are now restored to active participation and can now once again receive Microscopy Listserver Email. Sorry for the extended down time to those subscribers.
AOL in their infinite wisdom decided to block all Microscopy Listserver saying that it was a potential source of SPAM mail. A point that I would undoubtably argue to the contrary.
It has taken me several weeks to get them to reverse this decision and I had to jump through a lot of proverbial hoops. But as of this today mail appears to be flowing to blocked addresses at AOL.
I might point out that alot of ISP's, Companies and Universities have changed their SPAM software recently, and unilaterally block content or domains. For example I received rejected Email from several organizations from the (valid) Email posting asking about WebCams for microscopy. It was unilaterally decided that any Email message with the word Webcam was obviously SPAM mail. This was clearly not ture for this Email, but nevertheless the SPAM filters at various sites rejected it. This will be a continuing problem, for which I see little solution. So remember to choose your subject words carefully...
In any event, please don't be bashful of reporting outageous of MListserver Email which persist for several days. Your site may institute a domain or subject filter which blocks MListserver Email and I will not necessairly know about it.
For example,, AOL never actually notified me that they were blocking traffic. I only discovered it when testing a new filter and found that my AOL account was not receiving the test messages. That event (over 2 weeks ago) started me on search for the problem, which I erroneously thought I created, but happened to coincide with their system change.
BTW, I note with irony, that their blocking of the MListserver did not actually decrease the amount of SPAM mail I get on my (seldom used) AOL account, it only stopped me from using AOL, and also from my 80 year old Mother from receiving Email addressed from me via my normal microscopy.com account!
Sigh, now if I were running AOL things would be different.... ( Nestor grins )
The IFBLS'26 has been added to the list at the http://www.petr.isibrno.cz/microscopy/meetings.php#2004 . Other suggestions?
Petr Schauer
On 24 Sep 2003 at 7:51, Gareth Morgan wrote:
} That reminds me................ } International Federation of Biomedical Laboratory Science, IFBLS (formerly } IAMLT) will hold its 26th Congress in Stockholm, Sweden between the 13th } and 18th June, 2004. } } Take a look at: } http://www.vardforbundet.se/ifbls2004 } } } Med vänliga hälsningar/With best regards } } Gareth } } ***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look at } http://www.vardforbundet.se/ifbls2004 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } Gareth Morgan MPhil MSc FIBMS, } Department of Laboratory Medicine (Labmed), } Karolinska Institutet, } Huddinge Universitetssjukhus, 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. } }
Dear Members, I would like to try to observe cristalized KDP (KH2PO4) with a TEM and for this, I would like to obtain ultrathin sections of this cristalized KDP. However, I have no experience in cutting cristals with an ultramicrotome. Does anyone has ever try that? If so, what would be the best method? Thank you very much for any advice on this.
Pierre-Yves Sizaret
P-Y Sizaret Microscopy Facilities University of Tours, France
I noticed that you have PITTCON listed now... It might also be worth noting that we run the American Chem Soc short course, "Applied Optical Microscopy" in conjunction with that meeting. It will be held March 5-7 in 2004.
Thanks, Barbara Foster
At 10:59 AM 9/24/03 +0200, Petr Schauer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
P-Y Sizaret wrote: ====================================================== Dear Members, I would like to try to observe cristalized KDP (KH2PO4) with a TEM and for this, I would like to obtain ultrathin sections of this cristalized KDP. However, I have no experience in cutting cristals with an ultramicrotome. Does anyone has ever try that? If so, what would be the best method? Thank you very much for any advice on this. ====================================================== We have not tried cutting this particular crystal but we have thin sectioned , over the years, crystals that would seem to be not too different in properties.
You can not actually "section" with ultramicrotomy the entire crystal. Or at least we have never succeeded in doing that. But you can powder up some small amount of the crystal, we always would use a good mortar and pestle made from agate (not porcelain), see URL http://www.2spi.com/catalog/tools/mortar_pestles_agate.shtml and then embed the powder in one of the "Epon substitutes" such as our SPI- Pon 812 resin, see URL http://www.2spi.com/catalog/chem/spi-pon-812-epoxy-embedding-kit.shtml If you use a porcelain mortar and pestle, you will end up with porcelain powder in your ground sample, created a much more difficult time at interpreting your results.
If you experience "particle pull-out" then you have to either a) grind down to a smaller powder or b) use an adhesion promoter such as SPI-Chem™ 3GTMO, as shown on URL http://www.2spi.com/catalog/chem/trimethoxysilane.shtml
Since cutting crystals like this will be "hard"on your diamond knife, we would suggest as a money-saving measure, the use of a materials science diamond knife, but not one with a larger angle because compression effects would then make it much more difficult if not impossible to obtain usable sections of the particles (crystals). This is explained more fully on URL http://www.2spi.com/catalog/knives/materials.shtml This is a far more economical approach than to be using so-called "life science" diamond knives for this kind of work.
If you have other questions, let me know and I will try my best to answer them for you.
Disclaimer: SPI Supplies offers all three of the products mentioned in this posting so we would have a vested interest in having more people using these products.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
I am working with a colleague to examine isolated Chodium chloroplasts by TEM. The chloroplasts were isolated according to a published protocol (I can get those details if pertinent), and their final solution was 0.9 M sorbitol/0.05 M Hepes buffer, pH 7.3. Some but not all were pelleted into 2% agarose in the same solution prior to fixation in Karnovsky's. Following washes in 0.1M cacodylate + 7% sucrose, they were post-fixed in buffered 1% OsO4, then were washed, enbloc stained in UA, and routinely dehydrated and embedded (Embed 812). Thin sections were stained with UA and Pb, then examined in the TEM.
The same protocols yield excellent results for mammalian tissues. (That may be the key here?)
Some questions:
1 - Is it usual for the chloroplasts to remain a bright green throughout processing? There was no obvious sign of Os reduction (blackening of the tissues).
2 - Missing membranes? The outer limiting membranes of the chloroplasts are missing. Thykaloid membranes are fairly well-preserved (a few ripples here and there). Either the outer membranes are utterly unstained and thus invisible? or, more likely, stripped away during isolation although the stroma maintains a high degree of integrity and does not look extracted. The stroma looks reasonably dense, starch granules are dark and prominent.
Any explanation for the persistent green coloration? Any thoughts on our missing outer membranes?
Thanks all. Ann
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
Nestor, it never fails to astound me how ridiculous and anti-productive our modern life is getting. You're doing too fine and valuable a job to get mired in this kind of bureaucratic, er, um, stuff. My continued thanks for all your good work on our behalf. Dee
----------------------------------------------------------------------- } } Colleagues } } I believe AOL & Compuserve & Netscape.Net Email users are now restored } to active participation and can now once again receive Microscopy } Listserver Email. Sorry for the extended down time to those subscribers. } } AOL in their infinite wisdom decided to block all Microscopy Listserver } saying that it was a potential source of SPAM mail. A point that } I would undoubtably argue to the contrary. } } It has taken me several weeks to get them to reverse this decision and } I had to jump through a lot of proverbial hoops. But as of this today mail } appears to be flowing to blocked addresses at AOL. } } I might point out that alot of ISP's, Companies and Universities have changed } their SPAM software recently, and unilaterally block content or domains. } For example } I received rejected Email from several organizations from the (valid) Email } posting asking about WebCams for microscopy. It was unilaterally decided } that any Email message } with the word Webcam was obviously SPAM mail. This was clearly not ture for } this Email, but nevertheless the SPAM filters at various sites rejected } it. This will } be a continuing problem, for which I see little solution. So remember to } choose } your subject words carefully... } } In any event, please don't be bashful of reporting outageous of } MListserver Email which } persist for several days. Your site may institute a domain or subject filter } which blocks MListserver Email and I will not necessairly know about it. } } For example,, AOL never actually notified me that they were blocking traffic. } I only discovered it when testing a new filter and found that my AOL account } was not receiving the test messages. That event (over 2 weeks ago) } started me on search for the problem, which I erroneously thought I created, } but happened to coincide with their system change. } } BTW, I note with irony, that their blocking of the MListserver did not } actually } decrease the amount of SPAM mail I get on my (seldom used) AOL } account, it only stopped me from using AOL, and also from my 80 year old } Mother from receiving Email addressed from me via my normal microscopy.com } account! } } Sigh, now if I were running AOL things would be different.... ( Nestor } grins ) } } ;-) } } Nestor } Your Friendly Neighborhood SysOp.
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
I was hoping someone may have some experience with a building's primary power transformer (1500kW), and installing an SEM.
Presently we're concerned with the anticipated location being directly above this transformer, but the facility design is also in the design phase, and can be moved. One question would be how far(?), and another would ask if there aren't any extenuating circumstances(?)
While the entire building is being rennovated, I cannot imagine any ability for anticipating field strengths or measuring them.
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
Hi there, We would like to locate intermediate filaments in mouse pancreas using immunocytochemistry. The samples have been fixed using 4% PAF and 0.5% glutaraldehyde and embedded in LR White. They have been successfully used for other ICC localization. Could anyone recommend an antibody to use against proteins specific to mammalian intermediate filaments. We would prefer to find a polyclonal antibody if possible.
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
I am looking for a couple of sturdy tables on which to place microscopy systems. I know that excellent vibration isolation is acieved by products like TMC tables, however, can anyone recommend good (one level lower than excellent) isolation with other products?
Any personal experience or manufacturer name would be highly appreciated.
Thank you. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
Anyone care to comment on imaging samples in the SEM (LaB6) while they are offgassing naphthalene. The entomologists are still using the stuff and I have been kicking them out or transferring them to the ESEM, but is this really a problem for the instrument?? Since many of these samples are dried and completely unfixed they are offgassing all kinds of crap anyway, so should I even care? The policy was enforced as a result of the mess it makes of the sputter coater so I just banned it from the SEM's too. Maybe I don't need to though?? Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
Dear Michael, Last year we had a proposal to renovate the basement of the research building for a high-resolution EM and SIMS lab, so I used my Gauss Mauss to check the fields. This is a hand-held field checker that I bought from Marivac on sale a few years ago (I think it was about $300CAD). It has proved its worth many times over. You can get total field and AC component, check in horizontal and vertical directions and it is still working on its first set of batteries. When we had the room surveyed by one of the EM vendors, the results were very close. We were concerned because the building power supply cable vault was just under the floor, but because it was a high voltage feed, the field was negligible three to four feet away. The room is planned around that. You can also pay an EM vendor service department to do a survey of both mag fields and vibration, so there is no obligation. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, September 24, 2003 8:00 AM
Jean,
I have a vintage 1991 Ted Pella CPD. It is as manual as they come and after 12 years it still performs reliably.
Regards, Evelyn
Scripps Institution of Oceanography University of California, San Diego Analytical Facility (858)534-2438
-----Original Message----- } From: jean-ross-at-uiowa.edu [mailto:jean-ross-at-uiowa.edu] Sent: Monday, September 22, 2003 7:22 AM To: Microscopy-at-MSA.Microscopy.Com
Hi everyone,
I would like some opinions about critical point dryers. What do you think about automated units vs. manual units. Or do you use HMDS and not use the CPD at all? If you could post to the list and not send them directly to me, I would appreciate it and everyone could read the responses.
Thanks, Jean Ross Central Microscopy Research Facility University of Iowa
I attempted pre-embed immunogold labeling of neurofilament in rodent cochlea using 10 nm gold and did not have much success. While 6 nm is smaller than what I used, I would probably suggest going smaller to help avoid the problems I encountered. I have since gone to post-embed immunolabeling with the 10 nm gold and have had success.
Good luck!
On 23 Sep 2003 at 8:39, Robert Simmons 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 } ---------------------------------------------------------------------- } - } } Dear Microscopy Listserver members, } } I wonder if it is possible to use 6 nm-gold-tagged antibodies for } pre-embedding immunolabeling, or if it is necessary to use smaller } ones? I want to do immunolabeling of betacells from the islets of } Langerhans (rat). } } Sincerely, } Pernilla Nevsten } } } ...................................................................... } ..... .............................................................. } Pernilla Nevsten, PhD-student Materials Chemistry Lund University, } Sweden } } } } } I usually use something along the line of the Nanoprobes 1.4nm } nanogold for pre-embedding labeling, the cells are permeabilized after } primary fixation. You can enhance it after the labeling step (before } embedding) to make it easier to locate in the microscope. } } } Dr Robert Simmons } Program Director } Biological Imaging Core Laboratory } Georgia State University } Atlanta, GA 30302-4010 } } 404-651-3138 } 404-651-2509 FAX } } } }
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
I've been taking on a post-embed immunogold staining project and I'm having a low section adhesion average. My staining periods and rinses are quite long so there is ample opportunity for the sections to float off. My latest attempt had 1 grid out of 6 that retained its sections, and that's a poor average even if I'm a hitter for the Detroit Tigers!
A colleague emailed me the grid adhesive described by Bozzola, 2nd Ed but wonder if there are any drawbacks I need to consider, both with the beam and any known reactivity with the gold reagent?
Any suggestions to increase my success are welcomed!
Thank you
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
On Wednesday, September 24, 2003, at 10:49 AM, Scott Whittaker wrote:
} Anyone care to comment on imaging samples in the SEM (LaB6) while they } are } offgassing naphthalene. The entomologists are still using the stuff } and I } have been kicking them out or transferring them to the ESEM, but is } this } really a problem for the instrument?? Since many of these samples are } dried } and completely unfixed they are offgassing all kinds of crap anyway, so } should I even care? The policy was enforced as a result of the mess it } makes } of the sputter coater so I just banned it from the SEM's too. Maybe I } don't } need to though?? Thanks } Dear Scott, When I was doing electron diffraction on naphthalene in a TEM, I put the grid in a cryo-holder and started putting in LN2 while the airlock was pumping down. That was sufficient to prevent the naphthalene from subliming. I don't know how cold you would need to get your specimen--i.e., I can't say whether a Peltier cooler would do it--but at ~ -185 C there is no observable effect on the vacuum in a TEM from the nanoliter quantities I had on the grid, and there was no noticeable loss of the specimen from the grid, although I'm sure that a higher electron dose would etch naphthalene away. YMMV. Yours, Bill Tivol 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
On Wednesday, September 24, 2003, at 02:59 AM, Pierre-Yves Sizaret wrote:
} I would like to try to observe cristalized KDP (KH2PO4) with a TEM and } for this, I would like to obtain ultrathin sections of this cristalized } KDP. However, I have no experience in cutting cristals with an } ultramicrotome. Does anyone has ever try that? If so, what would be the } best method? } Thank you very much for any advice on this. } Dear Pierre-Yves, Do you need to observe the particular crystals you have, or would any KDP crystals do? If the latter, you could dry down a small volume of dilute solution--elevating the temperature will give smaller crystals. If the former, try to match the hardness of the resin to that of the KDP. Another concern is that if the substance is actually KH2PO4.xH2O, the structure can change in the TEM vacuum. Good luck. Yours, Bill Tivol 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
I am not familiar what KDP is. I did oriented (perpendicular to the crystallographic axes) 10 nm thick sections of the 3D ribosomal crystals and some other biological macromolecules. I used to process them in the pretty standard way with embedding into Epon. If crystals was big enough (0.2-0.5 mm) I orient them before polymerization. If the crystals are small (less than 0.1 mm) it's practically impossible to orient them - so you will have a set of different planes. Some people used tannic acid to improve the preservation of the crystals during the dehydration and embedding. I had perfect sections from the whole crystal without huge problems. We used those sections for the crystal structure determination. The crystal cell unit parameters obtained from the sections were in great agreement with X-ray crystallography data obtained later. EM also helped to distinguish between two group of symmetry X-ray guys had problem with. As a joke, I determined 2D cell dimensions for G-factor crystals in two days with very good preciseness (+/- 1A) as X-ray guys. Nevertheless it took them 2 years to prove that my data was correct. Ironically, by mistake, my EM images of the cell unit were presented as a X-ray solution. We also used EM for crystal's quality determination. Sergey
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
O-o-o-o-ps-s-s I just post reply about ribosomal crystals, which is nothing to do to inorganic materials. I apologize for not paying attention. Sergey
At 02:59 AM 9/24/2003, 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 33-089 Los Angeles, CA 90095
We have a Leica (formerly Wild) M650 surgical microscope on the floor stand. Beautiful instrument.
We would like to have an alternate stand so that the optics could be used on a table top.
I believe such a stand is used in the Leica M651 MSD. I would appreciate hearing from anyone who has information on this topic (vendors, users, etc.)
Thank you.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (brian.kirkmeyer-at-iff.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 24, 2003 at 09:15:33 ---------------------------------------------------------------------------
Email: brian.kirkmeyer-at-iff.com Name: Brian Kirkmeyer, Ph.D.
Organization: IFF
Title-Subject: Replication from skin for OM
Question: I am conducting an experiment in which a substance is applied to skin and washed away (like with soap), and we need to examine the residue remaining on the skin. My arm, which is the skin in question, does not fit in either of our optical microscopes, so I thought replication might be the way to go.
What is the best way to replicate the residue on the surface of my skin for analysis with OM? Is something simple like scotch tape the way to go? For the record, I would prefer to use benign substances on my skin...I like my arm and would like to keep it. :)
I'm not sure about your problem but we couldn't install SEM in the entire building which was located about 50m from the street train's stop. A change in the magnetic field was measured when the train was starting.
} Dear Ann, } } To your problems with the chloroplasts I can recall that here in } the lab we fix with glutaraldehyde 2 % in the same buffer (the same } molarity and pH) used for isolation. Post fixed in 2% Osmiun T.- The } Karnovsky is too much hypertonic for plants cells and tissues. The } chloroplast do not have to much lipid materials to reduce the Osmiun, } however 2 or 3 hours will rend enough to get a good conservation and } contrast.
In my hummble opinion! Have a good luck! Francisco
Francisco Freire, F.R.M.S. Head Service Electron Microscopy Service University Las Palmas de Gran Canaria Canary Islands Spain
} } } } Lehman, Ann wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You might try contacting Nuhsbaum Inc. They recently sold me a table top boom stand for my stereo microscope. web site is: http://www.nuhsbaum.com/
Best of luck,
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
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We have a Leica (formerly Wild) M650 surgical microscope on the floor stand. Beautiful instrument.
We would like to have an alternate stand so that the optics could be used on a table top.
I believe such a stand is used in the Leica M651 MSD. I would appreciate hearing from anyone who has information on this topic (vendors, users, etc.)
Thank you.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Amanda.Harman-at-newcastle.edu.au) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 25, 2003 at 00:30:17 ---------------------------------------------------------------------------
Organization: Reproductive Science Group, Newcastle University Australia
Title-Subject: Immunofluoresence with LR White Resin
Question: Hi,
I have been unsuccessfully trying to label 1 micron LR White resin sections of mouse epididymis,with a FITC tagged antibody. We are successfully gold labelling thin sections with the same antibody from the same blocks.
Brian Kirkmeyer wrote: ============================================== Title-Subject: Replication from skin for OM
Question: I am conducting an experiment in which a substance is applied to skin and washed away (like with soap), and we need to examine the residue remaining on the skin. My arm, which is the skin in question, does not fit in either of our optical microscopes, so I thought replication might be the way to go.
What is the best way to replicate the residue on the surface of my skin for analysis with OM? Is something simple like scotch tape the way to go? For the record, I would prefer to use benign substances on my skin...I like my arm and would like to keep it. :) =============================================== I can refer you to one of my own (with R. Swayne and T. Nightingale) publications:
Characterizing Cosmetic Effects and Skin Morphology by Scanning Electron Microscopy", presented at SCC Annual Educational Symposium, May 1975, St. Louis; J. Soc. Cosmet. Chemists, 27, 509-531 (November 1976).
I don't have any reprints left but I trust you would have this paper in the IFF library.
The detection and measurement of materials that are substantive to skin (meaning that they tend to stick to the skin instead of being washed off such as active moisturizer materials that might be incorporated into a bar of soap) is not a straight forward analytical undertaking. We have found that if one can be happy with measuring the effect of the deposited material , rather than the material itself, if the work is done on cosmetically dry skin, and if you are doing replicas of the skin site (with the right replicating material), before vs. after product application, when there is a deposition of something, its effect can be measured by careful comparision with the before replicas. One can't do this kind of work on "normal" skin because there would not be present the uplifting (stratum corneum) layers from which one could measure such changes. The results can be analyzed semi-automatically with image analysis.
Although they are not images from the above referenced paper, examples of such skin replicas can be seen on URL http://www.2spi.com/catalog/spec_prep/a.html
These replicas were made with the SPI-Chem™ Wet Replica kit which uses a fast acting curing agent so that the true skin morphology is captured, and not that of a highly hydrated skin site (which would be the result if the replicating resin was curing over a longer time frame such as several minutes). See URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.shtml
Scotch tape strippings and even cyanoacrylate strippings (which do "hurt" when stripped off) don't seem to be very helpful in terms of measuring the amount present. We have tried measuring quantitatively, the amount of residue remaining by taking multiple Scotch tape strippings, each one removing a bit more of the residue, so that when all of the residue was removed, that last stripping really did "hurt". Counting the number of strippings needed to get down to that point, in theory at least, one could make some determination about the amount of residue present. But we could never get this approach to work very reproducibly either within our own laboratory or between laboratories using the same product application protocol.
Disclaimer: SPI Supplies is the manufacturer of the SPI-Chem Wet Replica Kit. The independent laboratory of Structure Probe, Inc. has been performing these kinds of studies for clients in the cosmetic and pharmaceutical industries since 1972.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
Nestor, I agree keep up the good work. Now a little problem, for the last week or so all the communications I receive from the listserver comme in duplicate. Isi this a problem on the server or is my e-mail package doing this to me? Has anyone else had this problem.
Thanks,
Ron.
Dee Breger 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 } ----------------------------------------------------------------------- } } Nestor, it never fails to astound me how ridiculous and anti-productive our } modern life is getting. You're doing too fine and valuable a job to get } mired in this kind of bureaucratic, er, um, stuff. My continued thanks for } all your good work on our behalf. Dee } } ----------------------------------------------------------------------- } } } } Colleagues } } } } I believe AOL & Compuserve & Netscape.Net Email users are now restored } } to active participation and can now once again receive Microscopy } } Listserver Email. Sorry for the extended down time to those subscribers. } } } } AOL in their infinite wisdom decided to block all Microscopy Listserver } } saying that it was a potential source of SPAM mail. A point that } } I would undoubtably argue to the contrary. } } } } It has taken me several weeks to get them to reverse this decision and } } I had to jump through a lot of proverbial hoops. But as of this today mail } } appears to be flowing to blocked addresses at AOL. } } } } I might point out that alot of ISP's, Companies and Universities have changed } } their SPAM software recently, and unilaterally block content or domains. } } For example } } I received rejected Email from several organizations from the (valid) Email } } posting asking about WebCams for microscopy. It was unilaterally decided } } that any Email message } } with the word Webcam was obviously SPAM mail. This was clearly not ture for } } this Email, but nevertheless the SPAM filters at various sites rejected } } it. This will } } be a continuing problem, for which I see little solution. So remember to } } choose } } your subject words carefully... } } } } In any event, please don't be bashful of reporting outageous of } } MListserver Email which } } persist for several days. Your site may institute a domain or subject filter } } which blocks MListserver Email and I will not necessairly know about it. } } } } For example,, AOL never actually notified me that they were blocking traffic. } } I only discovered it when testing a new filter and found that my AOL account } } was not receiving the test messages. That event (over 2 weeks ago) } } started me on search for the problem, which I erroneously thought I created, } } but happened to coincide with their system change. } } } } BTW, I note with irony, that their blocking of the MListserver did not } } actually } } decrease the amount of SPAM mail I get on my (seldom used) AOL } } account, it only stopped me from using AOL, and also from my 80 year old } } Mother from receiving Email addressed from me via my normal microscopy.com } } account! } } } } Sigh, now if I were running AOL things would be different.... ( Nestor } } grins ) } } } } ;-) } } } } Nestor } } Your Friendly Neighborhood SysOp. } } *************************************************************** } Please do not publicly post any of my correspondence without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995)
-- Ronald J. Smith Department of Biology Room 235, Biological & Geological Sciences Bldg. U.W.O., London, Ontario N6A 5B7 (519) 661-2111 ext.86486
We routinely use grid pens from Electron Microscopy Sciences, due to terrible problems with losing sections and having sections tear apart during incubations for immuno work. This adhesive has almost completely solved our problems and even appears to have improved section stability under the beam.
As far as any effect on labeling, I have never done any side-by-side comparisons with and without adhesive, but I have not noticed any obvious effects on our labeling.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---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.biotech.missouri.edu/emc/
-----Original Message----- } From: Christopher S. Zurenko [mailto:czurenko-at-mail.khri.med.umich.edu] Sent: Wednesday, September 24, 2003 4:33 PM To: Microscopy-at-MSA.Microscopy.Com
I've been taking on a post-embed immunogold staining project and I'm having a low section adhesion average. My staining periods and rinses are quite long so there is ample opportunity for the sections to float off. My latest attempt had 1 grid out of 6 that retained its sections, and that's a poor average even if I'm a hitter for the Detroit Tigers!
A colleague emailed me the grid adhesive described by Bozzola, 2nd Ed but wonder if there are any drawbacks I need to consider, both with the beam and any known reactivity with the gold reagent?
Any suggestions to increase my success are welcomed!
Thank you
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
} Question from: } } Christopher S. Zurenko } Kresge Hearing Research Institute, Otopathology } The University of Michigan Medical School
} I've been taking on a post-embed immunogold staining project and I'm } having a low } section adhesion average. My staining periods and rinses are quite } long so there is } ample opportunity for the sections to float off. My latest attempt } had 1 grid out of 6 } that retained its sections, and that's a poor average even if I'm a } hitter for the Detroit } Tigers! } } A colleague emailed me the grid adhesive described by Bozzola, 2nd } Ed but wonder } if there are any drawbacks I need to consider, both with the beam } and any known } reactivity with the gold reagent?
Comments:
Poor adhesion of sections is generally due to either dirty grids or grids that are not flat (bent or wavy). Before using grid glue, try cleaning the grids by sonication in a solvent such as acetone. Alternatively, you might use an acid such as 1M HCl for 5-10 minutes. If cleaning alone does not solve the adherence problem, then place the grids onto a filter paper (with the side that will hold the sections facing up) and place droplets of 0.25% Formvar or Collodion onto the grids. This puts a plastic coating over the meshwork of the grids without obstructing the open areas. The plastic coating enhances adherence of the plastic sections significantly. Also, after the sections are put onto the grids, place them overnight in a 60 C oven prior to immuno staining. This works wonders.
If all of these fail, maybe then I would consider grid glue as the last resort. But, really, you should not need it.
Let us know what solves your problem.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
In a message dated 9/25/03 12:02:40 PM, rsmith-at-uwo.ca writes:
} I agree keep up the good work. Now a little problem, for the last week } or so all } the communications I receive from the listserver comme in duplicate. Isi } this a } problem on the server or is my e-mail package doing this to me? Has anyone } else had } this problem.
This happens to me as well, but (strangely) only on about half of the messages, and the second copy arrives quite some time after the first one. I was blaming my email client, since I have messages being forwarded from one address to another, but have no real explanation. John Russ
There are several possible explainations for duplicate messages.. If this problem persists by all means contact me, but don't post this problem to the MicroscopyListserver, send an off-line message to me explaining the problem and if at all possible include an example. and I'll try to help you sort it out.
Here are a few tips, if you would like to do some detective work ahead of time.
1.) If every message you receive is a duplicated and this persists for a long time (i.e. days) then it is likely you actually receiving multiple seperate mailings. There can be 2 possibilities here
a.) You are really subscribed multiple times.
The important information is in your Email header are your two subscriptions both the same email address or not. Some people have inadvertently subscribed with similiar (yet different) addresses..
These may all be you, but the server treats these as 3 different addresses and will dutifully sent out 3 copies of the message, arent' computers wonderful, they do what you tell them not what you mean't. Check your mail headers and if you are receving posts at different addresses unsubscribe the one you don't want. Use the WWW based form at
b.)Someone is (or has started) forwarding mail from their address to you, or you are forwarding mail to yourself from another system.
This happens sometimes when, for example, an employee leaves a company and email is forwarded from say an info-at-xyz.com account to another person in the company. Again you need to check your header lines to see how the mail is getting to you.
In both cases above I'll try to help, but I'll also need the header info to sort it out (at least to sort it out more easily).
2.)Occassional duplicate messages...
a). Check the time stamps in the headers
If the time stamps are different then it is likely that the sender just mailed it twice. This happens occassionally
b.)Time stamps identical, but duplicate messages are an occasional occurance.
Likely due to a mail delivery problem. All mail servers attempt to insure delivery. If an error is detected during the sending process the mail server may resend the whole message. The severity of the error is also sometimesirrelevant, it could just be a missing character even a space at the end of the message. It will be resent depending upon the error codes sent back an forth between the mail delivery agents (MDA's). This can happen because of network failures, congestion and or server errors. This problem can occur anywhere along the transmission path. It could be at this server, a relay point along the net, your ISP's server, or your client program having problems connecting and requesting a download to your local computer.. Unless this persists for a long time, there is not much that can be done, since the error is probably not reproducible.
3.) The whole list starts getting duplicates... I'll catch this, (it has happened a few times) but remember even I ocassionally have another life, and I might not catch the problem until later in the evening when I do system checks.
Also remember that I closely manage the subscription and unsubscription process so that spam/junk mailers have a difficult time getting in. New subscriptions and unsubscriptions must be approved, i.e. it is not an automatic process. This is done only once/day, so don't expect instanteous changes.
This minimizes junk mailers getting into the system, but it does delay things.
I'll also remind you that it takes time for all the mail queues to process. Postings don't happen immediately, there are delays in processing and delivery. On average the delay is on the order of a few hours. If you attempt to post a message and you don't see it within a day something is likely wrong.
All rejected messages which are spam suspects are automatically sent back to the sender with an explaination of the problem. However, I note that lately, some sites are flagging the MListserver warning messages as SPAM. It's a catch 22 situation and there have been a few subscribers caught in the middle with both servers rejecting each others Email, meaning the person who posted the message is out of the loop and doesn't know what is happening. In this case (i.e. you don't see your posting after about 24-48 +hours), contact me off-line or try using the on-line WWW submission form also at
as with the subsription WWW page this one is closely monitored to limit spam and there will be delays since these forms requires my personal "blessing" to be forwarded to the list.
Dear Colleagues, we are searching for an antibody to analyze mitotic activity in mouse tissue (cryo sections, formaldehyde-prefixed, 4% pFA). Does anybody have recommendations for a nice ki67 or pcna (proliferating cell nuclear antigen) antibody to detect proliferation? Any experiences, tricks, tips and recommendations are welcome.
Thanks in advance
Daniel
-- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V=
Over the weekend I have modified the Microscopy Listserver Email filters which I hope should help you all in the constant SPAM/Virus battles.
As you know there is alot of Address spoofing (i.e. faking) going on lately, where a false sender's address is inserted into an Email to make you think it is coming from somewhere different than the actual source.
For many years the MListserver has been inserting a header line into the body of all Emails which indicates it is being sponsored by the Microscopy Society of America, and with a bit of reminder info on how to subscribe/unsubscribe. This is all handled by the SPAM filter, and was intended to help you weed out junk mail.
Many people filter email based upon the content of Email headers and/or the Email text in the message body. I have modified the SPAM filters so that all Email which now goes through the Filter now also should have it's SUBJECT modified and the phrase
[MicroscopyListserver]
prepended to the actual text of the original subject. This should help you all differentiate spoofed email from items that have actually been processed by the ML filter.
This makes the Subject line abit longer, but it adds abit of insurance that the message "might" be real ;-)
Of course, I have every confidence that this will be defeated at some time in the not to distant future, but it should help for awhile.
Remember, just because an Email appears to be coming from Microscopy.Com, you should not trust it implicitly. There was a virus sent out at the end of last week purportedly about Microscoft security updates. Had this gone through the ML Filter it would have been blocked because of the attachment which it contained.
The battles never end...
Cheers
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Sun Sep 28 16:31:04 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (docM_ph-at-hotmail.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, September 27, 2003 at 00:31:12 ---------------------------------------------------------------------------
Email: docM_ph-at-hotmail.com Name: Martin Moreno
Organization: St} Luke's Medical Center
Education: Graduate College
Location: Quezon City,Philippines
Question: Im trying to visualize viruses by Negative Stain on a transmission EM. BUT,I cant seem to get the right stain. Casn you please help me on this? Id appreciate it very much. Thanks you in advance.
I am looking for any used, but operational equipments, either as a donation or with a small charge. The equipments include: Microscopes (LOM, TEM, SEM, SPM) HPLC DSC/TGA FTIR GC-MS AA Thanks! Pat Sadhukhan
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 10:32:04 2003
The final notice for the Midwest Microscopy & Microanalysis Society (MMMS) Meeting in Madison, Wisconsin on October 3rd, can be found at the following URL: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSOct2003.pdf
The MMMS Society in association with the ASM Chicago Chapter is holding an SEM workshop at Harper College in Palatine, Illinois on November 14th. The first announcement for this meeting can be found at the following URL: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSNov2003.html
For further information please contact either:
Robert Mierzwa (MMMS President - Elect): Tel: 920-803-8945, email: mierzwa-at-jeol.com Arvid Casler (MMMS Program Coodinator): Tel: 847-674-7700, email: arvid_casler-at-fmo.com Jim DiOrio (MMMS President): Tel: 847-270-4676, email: jim_diorio-at-baxter.com
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 11:52:26 2003
The Connecticut Microscopy Society announces plans for its Fall Meeting October 23rd at Wesleyan University in Middletown, CT, featuring:
Dr. David Hall, from Albert Einstein School of Medicine. TITLE: "The Complete Nematode: Exploring C. elegans Tissues by EM"
Dr. Ron Anderson, former MSA President and now Editor of Microscopy Today. TITLE: "TEM Analysis of Thin Films: Specimen Preparation"
Colleagues
It was pointed out to me that the [MicroscopyListserver] label that I added over the weekend was far too long, and upon reflection I must agree. Chalk it up to too many hours in front of a computer screen.
I've shortened it to just [Microscopy]. Let's see how well this works.
Cheers...
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 14:39:50 2003
I am looking for recommendations for oxygen sensors. We have some small rooms where liquid nitrogen is used in amounts that require some safety measures including an oxygen monitor. I have found units ranging from about $600 to $1800 with varying costs and 1 year to 2 year lifetimes for the active sensor.
Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140, Room S101 San Francisco, CA 94143
415-476-4441 FAX 415-514-4142
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 19:18:23 2003
I would suggest "ListServ" or "ListServer" is better, because "Microscopy is too much common word and may not be effectively used in any filters. For instance, "electron microscopy" would be filtered as a message from listserver even if it's not. Another suggestion: if it possible, it would be better to add it to the end of the subject line, otherwise, I have difficulties to read subject line, it's mostly occupied by "Microscopy"... Thanks, Sergey
At 10:20 AM 9/29/2003, 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 33-089 Los Angeles, CA 90095
I think that the current [Microscopy] preface is very good. "ListServer" is too generic...there are at least 15,000 of them. A filter should only look at the Subject line. If it wants to dig deeper, it can look at the sub-headers. But for Nestor's purpose, and ours, I think that [Microscopy] is a great advancement. Plus, this keeps me from wrongfully posting to the XL-30 ListServer.
Thanks Nestor.
gary g.
At 05:21 PM 9/29/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 23:08:31 2003
I would agree that [Microscopy] would be more appropriate than [Listserver]; I think Sergey's idea of putting the tag at the end of the subject line instead is a good one, though. Sergey: for purposes of filtering, why not include the [] brackets in the string to match for the filter? Any thoughts/comments on using [MSA], or is that getting carried away?
------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
} } } ------------------------------------------------------------------------------ } 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 that the current [Microscopy] preface } is very good. "ListServer" is too generic...there } are at least 15,000 of them. A filter should only } look at the Subject line. If it wants to dig deeper, } it can look at the sub-headers. But for Nestor's } purpose, and ours, I think that [Microscopy] is a } great advancement. Plus, this keeps me from } wrongfully posting to the XL-30 ListServer. } } Thanks Nestor. } } gary g. } } } } At 05:21 PM 9/29/2003, 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 } } ------------------------------------------------------------------------------- } } } } I would suggest "ListServ" or "ListServer" is better, because "Microscopy } } is too much common word and may not be effectively used in any } } filters. For instance, "electron microscopy" would be filtered as a } } message from listserver even if it's not. Another suggestion: if it } } possible, it would be better to add it to the end of the subject line, } } otherwise, I have difficulties to read subject line, it's mostly occupied } } by "Microscopy"... Thanks, Sergey } } } } At 10:20 AM 9/29/2003, 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 } } } ------------------------------------------------------------------------------- } } } } } } Colleagues } } } } } } It was pointed out to me that the [MicroscopyListserver] label that } } } I added over the weekend was far too long, and upon reflection I must agree. } } } Chalk it up to too many hours in front of a computer screen. } } } } } } I've shortened it to just [Microscopy]. Let's see how well this works. } } } } } } Cheers... } } } } } } Nestor } } } Your Friendly Neighborhood SysOp } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } 10833 Le Conte Ave, Room 33-089 } } Los Angeles, CA 90095 } } } } Phone: (310) 825-1144 (office) } } (310) 206-1029 (Lab) } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 10:59:29 2003
In tracking the logs (and my email) for the last few days I am starting to see that an increasing a number of postings are being rejected. Rejections in themselves are not a new event as it happens alot.
This is happening more and more often, because of the fact that people have changed their addresses or have a different return address than their sending address. The problem is that the warning message which is telling the sender that there is a possible problem with their post (and is automatically sent out by the Listserver ) are being rejected by many receipents hosts.
This means, of course, that the sender doesn't know there is a problem.
If you are having a problem posting a message, for the sort term please use the WWW based form at:
Sorry for the hassel, I'll investigate this latest problem and try to sort it out.
Nestor Your Friendly Neighborhood SysOp
-- ---===[|]===--- Nestor J. Zaluzec
Email: Zaluzec-at-Microscopy.Com {--------at- Home
Email: Zaluzec-at-aaem.amc.anl.gov {-------at- Argonne National Lab.
---===[|]===---
--------------------------------------------------------- The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac ---------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 12:05:33 2003
We have replaced the Link eXL (now Oxford) automation that controlled our Cameca SX50, and the computer and its peripherals can be made available to anyone interested and who can pay the packing and shipping charges. I imagine there may be someone who may need it for parts(?)
I, personally, do not know a whole lot about this automation for the SX50, but it has controllers for 3 WDX, a second terminal (no monitors), PC-Link software/hardware, and EDX control (we are keeping the detector). It would be too much work to cannibalize, so please do not ask.
Those interested should contact me direct ... {michael -at- shaffer.net}
Hi all, I am looking for a non-autoflourescent resin. I am labeling specific cells that are not synchronized. The cells grow on ACLAR, and they are fixed with glut, and OsO4, embedded on a piece of ACLAR with the cells facing down into the resin. This creates a thin layer of resin with cells close or directly on the surface. I'm using DAPI to stain the nucleus / chromosomes to identify cells of interest. I can see cells (although it is had to focus) using a light microscope, but the resin seems to be auto-fluorescent. Currently i have only tried Epon-812. Has anybody tried this before? Thanks in advance.
Joel Mancuso Department of Molecular and Cell Biology 345 LSA University of California Berkeley, CA 94720-3200
p - 510.643.8277 email - jmancuso-at-uclink.berkeley.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 12:50:17 2003
We have the DuoScan T2500 and the computer it was on died (Macintosh OS 8.?), so we tried reinstalling it on a different computer (Macintosh (9.1).
We cannot get it to scan higher than 600 PPI. Before it died we could scan at 2500 PPI.
Any help in installing/reinstalling to drive it at full resolution would be great.
Thanks.
____________________________________________________________________________ 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/
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 15:06:07 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yan.328-at-osu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 30, 2003 at 10:55:27 ---------------------------------------------------------------------------
Email: yan.328-at-osu.edu Name: Yanping Yan
Title-Subject: [Microscopy] EM, need help with serial reconstructions
Question: Hi, all:
We are attempting serial reconstruction the axon of a cultured neuron. The axons are just about 200 nm thick.
The problem that we have run into is that when we try to line up the axons in the two sections using unambiguous landmarks, such as large organelles or varicosities (which are large enough to be present in both sections), they simply don't line up. In the one case that we have examined so far, the distance between the two landmarks is about 18% greater in one section than in the other. This is definitely not due to errors in our montaging, because we have confirmed the discrepancy within single images of the two sections taken at low mag.
Some possibly relevant technical info is that we are embedding in a pretty standard PolyBed 812 formulation and cutting silver sections.
We guess the sections must have been compressed or expanded differentially during processing. What surprises us is that the discrepancy is quite large. Moreover, we would actually expect the amount of compression to be greatest perpendicular to the knife edge, yet the axon is actually running close to parallel to the knife edge (maybe about 30 degrees off parallel) in this case.
Do you ever encounter this problem when you were doing serial reconstructions? Do you do anything to avoid this problem? If we stretch the sections with chloroform or heat, we don't how much it will help since sections can react differently.
I am wondering if you can give us some suggestions.
Thanks,
Yanping Yan
Neuroscience Graduate Studies Program Rightmire Hall, Room 056 The Ohio State University Center for Molecular Neurobiology 1060 Carmack Road Columbus, OH 43210
Greetings Joel, Do you need to achieve good image quality for fluorescence or are you just screening for cells to perform EM analysis on? I'm not sure what the refractive index of Epon-812 is, and refractive index will have an impact on image quality. Glutaraldehyde is known to cause extensive background fluorescence, but you can probably still discriminate DAPI from the greenish glutaraldehyde signal. You might try epon/araldite as outlined in:
Sun, Tolbert and Hildebrand. (1995). Using laser scanning confocal microscopy as a guide for electron microscopic study: a simple method for correlation of light and electron microscopy. J. Histochem. Cytochem. 43(3),329-355.
Other methods to correlate images from photonic microscopy and electron microscopy include the use of cryosections, light microscope imaging in agarose followed by embedding for EM. Pertinent references are below:
Robinson, Takizawa, Pombo and Cook. (2001). Correlative Fluorescence and Electron Microscopy on Ultrathin Cryosections: Bridging the Resolution Gap. J. Histochem. Cytochem. 48, 471-480.
Sun, Tolbert, Hildebrand and Meinertzhagen. (1998). A Rapid Method for Combined Laser Scanning Confocal Microscopic and Electron Microscopic Visualization of Biocytin or Neurobiotin-labeled Neurons. J. Histochem. Cytochem. 46, 263-274.
Deitch, Smith, Swann and Turner. (1991). Ultrastructural investigation of neurons identified and localized using the confocal scanning laser microscope. J. Electron Microsc. Tech. 18, 82-90.
Sun, Tolbert and Hildebrand (1997). Synaptic organization of the uniglomerular neurons of the antennal lobe of the moth Manduca sexta: a laser scanning confocal and electron microscopic study. J. Comp. Neurol. 379,2-20.
Regards, Karl G.
jmancuso wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 17:44:30 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (beth-at-plantbio.uga.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 30, 2003 at 16:16:36 ---------------------------------------------------------------------------
Question: Hi all, We are trying to cryostat section root-knot nematodes and soybean cyst nematodes that have been fixed in 2% paraformaldehyde/PBS, etc. but even with a new blade in place the 'todes and roots are often tearing up rather than sectioning. Any help/advice would be greatly appreciated. We'd like to see other people's protocol for fixation and hear any thoughts on 10% DMSO vs 30% sucrose? We would esp. like to know how long you leave the samples in each step of your protocol. thanks!!! Beth
Question: I need info on microscopist as a career. I want a job description and the training needed. The different types or specialties would be great also. Please include other info that would be great for a research paper.