My wife had what appears to be a similar problem after a probably not so routine injection in the early 1980s. The injection was for protection from anthrax, which ironically in the end she never worked with. Although this has perhaps similarity with 'Gulf war Syndrome' 20 years later, her reaction to the injection seems very unlikely to be due the 'pathogen', more likely the carrier medium. She got an intense reaction at the site within 24 hours that remained very painful (couldn't be touched|) for a year or so, although the pain became intense and then reduced a little over weekly cycles. We don't think anything like steroid injections were tried as my wife certainly wasn't keen on any more needles, but it is a while ago now and she can't remember all the minor details (they must be in her notes).
As she describes below the only thing that got rid of it was a fairly major operation to remove the entire reaction site. It caused so much pain, although it got very bad and then a bit better in the weekly cycles, that she insisted on the operation to excise the area. As my wife mentioned, the first operation didn't remove enough of the inflamed area and the second probably removed slightly more than was necessary. However it was very successful in the end. She got no compensation or anything even though it was a work related injury, as nothing was really found (still seems a bit like Gulf War Syndrome) - the 'lump' was removed in a non-painful part of the cycle so there was no useful microscopy.list.server related histology. At the time such reactions seemed rare and little was known about them - it didn't even have a name (now probably all sorts of similar conditions are bundled under the 'macrophagic myofasciitis' label).
She had a minor complication due the resulting very large 6 inch vertical scar at the top of her left arm being 'wired' after the operation. A visiting locum doctor told her to change the rest position of the arm from that advised by the surgeons and it badly split the wound's stitching. The area of her arm now has a large sensitive scar (from the operation scar tissue) that hurts her if anyone pokes it and occasionally she gets the odd tingle from the old area near where the injection site was (probably scar tissue again), but nothing like she was suffering previously. Her arm clearly has a large dip where so much muscle tissue was removed, but it doesn't notice now she's older (53), although she tends to wear short sleeves rather than tank tops. Sunburn irritates it was well. However in comparison with the pain she suffered for a year or so when the inflamed area was present, she considers the operation was a complete success. This was all back in 1982, and medical staff here were non-plussed by the injection reaction at the time. She didn't get any systemic problems she was aware of, just around the injection site (although that was so painful it may have masked other symptoms I suppose). It was quite a drastic cure but it worked, and of course she went on to marry a great guy and have wonderful children while still juggling a successful career etc....
My wife's comments :
"They didn't treat it with anything. The only other cases I came across then (as it was pre-internet) were either allergy to the aluminium used in preparing the inoculum or in the 3rd world women reacted to the short chains of an 'oil' in the adjuvant which normally used long-chain molecules but not the inoculum itself.
Whatever it was, it was as instant as the day after the injection I could not lift or touch my arm.
Mine flared up every 2 weeks eventually like clockwork.
The first local excision did work but because they cut it out when it was 'quiet' they could not discern where it was. I told them they had cut it in half but of course they didn't believe me. So when it re-appeared on schedule 2 weeks later they decided to cut a huge amount out just to be sure. If they had done the 2nd op first it would not have needed to be so drastic but still sizeable."
I hope this is of some help.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Good afternoon, good evening, } dear listers, } } Allow to ask a question perhaps not usual for this forum: } } We do have a patient suffering from several pain due to a post-vaccination } problem (due to accumulation of Al-precipitates in intermuscular } macrophages) , namely } } macrophagic myofasciitis, } } luckily NOT systemic or generalized. } } If there is anybody out there who has knowledge about a specific and } reliable treatment for such a condition I greatly should appreciate your } comment. } (Unfortunately I was not able to localize any reference for } successful { } treatment). } } } If -perhaps- I have broken rules for using this listserver, please } apologize. It is just to seek help for a young patient. } } Thank you and } cordially yours } } Wolfgang Muss, PhD. } Salzburg, Austria }
==============================Original Headers============================== 16, 28 -- From keith.morris-at-ucl.ac.uk Wed Mar 1 04:17:37 2006 16, 28 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) 16, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21AHZEt004779 16, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 04:17:36 -0600 16, 28 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 16, 28 -- by vscani-d.ucl.ac.uk with smtp (Exim 4.51) 16, 28 -- id 1FEOOh-0007P3-Ns; Wed, 01 Mar 2006 10:17:31 +0000 16, 28 -- Message-ID: {003f01c63d19$3f7b26b0$7b865290-at-keithhigrade} 16, 28 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 16, 28 -- To: {Microscopy-at-microscopy.com} 16, 28 -- Cc: {W.Muss-at-salk.at} 16, 28 -- References: {200602271813.k1RID3wk015939-at-ns.microscopy.com} 16, 28 -- Subject: Re: [Microscopy] Vaccination complications: macrophagic myofasciitis 16, 28 -- Date: Wed, 1 Mar 2006 10:16:47 -0000 16, 28 -- MIME-Version: 1.0 16, 28 -- Content-Type: text/plain; 16, 28 -- format=flowed; 16, 28 -- charset="iso-8859-1"; 16, 28 -- reply-type=original 16, 28 -- Content-Transfer-Encoding: 7bit 16, 28 -- X-Priority: 3 16, 28 -- X-MSMail-Priority: Normal 16, 28 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 16, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 16, 28 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 16, 28 -- X-UCL-MailScanner: Found to be clean 16, 28 -- X-UCL-MailScanner-SpamCheck: 16, 28 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk ==============================End of - Headers==============================
I've used sputtered Ag on one occasion with some success. The only catch was that after sputtering, the samples went immediately into the scope, and once they were removed from the scope were not analyzed again. I didn't do any experiments to prove this, but I assumed the Ag would oxidize rather quickly. The Ag didn't do as good of a job of dissipating surface charge on the sample as Au would have, but it was adequate for the work I was doing.
Cheers, Bryan Bandli Research Microscopist MVA Scientific Consultants
gary-at-gaugler.com wrote:
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==============================Original Headers============================== 6, 19 -- From bbandli-at-mvainc.com Wed Mar 1 08:13:24 2006 6, 19 -- Received: from smtp05.gnvlscdb.sys.nuvox.net (smtp.nuvox.net [64.89.70.9]) 6, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21EDNut017324 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 08:13:24 -0600 6, 19 -- Received: from [192.168.1.70] (216.215.228.34.nw.nuvox.net [216.215.228.34]) 6, 19 -- by smtp05.gnvlscdb.sys.nuvox.net (8.12.11/8.12.11) with ESMTP id k21EDXHQ015464 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 09:13:33 -0500 6, 19 -- Message-ID: {4405AB92.6050208-at-mvainc.com} 6, 19 -- Date: Wed, 01 Mar 2006 09:11:30 -0500 6, 19 -- From: bbandli {bbandli-at-mvainc.com} 6, 19 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- MIME-Version: 1.0 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- Subject: Re: [Microscopy] Low Z peak pileup 6, 19 -- References: {200603010337.k213btxx031706-at-ns.microscopy.com} 6, 19 -- In-Reply-To: {200603010337.k213btxx031706-at-ns.microscopy.com} 6, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I sometimes use chromium or nickel to coat biological samples for EDS analysis. Cr can either be evaporated from Cr chips in vacuum evaporator, or sputtered from Cr target in sputter coater.The K-shell x-rays don't overlap any elements of biological interest and the L-shell is very low at about 0.57 KeV. Nickel L-shell is at about 0.89 KeV.
My experience with carbon coating is that it is not very good at eleiminating charging on highly insulating biological samples (critical point dried, freeze-dried) and also not being a "metal" is not a good source of secondary electrons for imaging. Bot Ni and Cr are very effective at eliminating charging and make for stable secondary electron images for capturing good images of what you are doing EDS analysis on.
Having said that, I shall attempt carbon coating today on biological sample to compare with EDS done on Cr coated sample a few weeks ago, so see if detectibility of trace amounts of Cu and Zn is improved.
Gib -- Gilbert (Gib) Ahlstrand, Electron Microscopist Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
-------- Original Message --------
Dear Keith, dear Phil (Oshel), dear Robert (Darby), Dear Diane (Ciaburry), dear Diane (Miller), dear listers, apologize for my perhaps rel. late reply to all kind opinions and/or messages I got concerning the header } Vaccination complications: macrophagic myofasciitis { (no other messages received except via MSA-Listserver).
I would like to thank you for your input and interesting/valuable comments, now knowing that there is not much new concerning an efficient treatment.
Yes, I have found also the reference on } one successful treatment { by a 2 years medication with steroids and azathioprine (Fischer,Reimann,Schroeder: Dtsch Med Wochenschr. 2003 Oct 31;128(44):2305-8) but in that case perhaps the myophagic "reaction" was not initiated by aluminium and therefore might have been caused by another constituent/circumstance .....the patient in our case unequivocally had Al in the macrophages' cytoplasm (as confirmed not only by typical EM-micrographs but now also by EDX and EELS) received corticosteroid treatment for appr. half a year without any improvement.
Chelating therapy for aluminium in human seems to be a little bit complicated perhaps, as I have seen from a lit. search, but perhaps I have located ONE person at Mount Sinai School of Medicine some minutes before. If you like, I'd send info directly / off Listserver to you provided that communication thread can be verified.
Also, I shall contact you personally within 24 h......(:-)) Have a nice and beautiful - great- day,
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Dear Wolgang,
My wife had what appears to be a similar problem after a probably not so routine injection in the early 1980s. The injection was for protection from anthrax, which ironically in the end she never worked with. Although this has perhaps similarity with 'Gulf war Syndrome' 20 years later, her reaction to the injection seems very unlikely to be due the 'pathogen', more likely the carrier medium. She got an intense reaction at the site within 24 hours that remained very painful (couldn't be touched|) for a year or so, although the pain became intense and then reduced a little over weekly cycles. We don't think anything like steroid injections were tried as my wife certainly wasn't keen on any more needles, but it is a while ago now and she can't remember all the minor details (they must be in her notes).
As she describes below the only thing that got rid of it was a fairly major operation to remove the entire reaction site. It caused so much pain, although it got very bad and then a bit better in the weekly cycles, that she insisted on the operation to excise the area. As my wife mentioned, the first operation didn't remove enough of the inflamed area and the second probably removed slightly more than was necessary. However it was very successful in the end. She got no compensation or anything even though it was a work related injury, as nothing was really found (still seems a bit like Gulf War Syndrome) - the 'lump' was removed in a non-painful part of the cycle so there was no useful microscopy.list.server related histology. At the time such reactions seemed rare and little was known about them - it didn't even have a name (now probably all sorts of similar conditions are bundled under the 'macrophagic myofasciitis' label).
She had a minor complication due the resulting very large 6 inch vertical scar at the top of her left arm being 'wired' after the operation. A visiting locum doctor told her to change the rest position of the arm from that advised by the surgeons and it badly split the wound's stitching. The area of her arm now has a large sensitive scar (from the operation scar tissue) that hurts her if anyone pokes it and occasionally she gets the odd tingle from the old area near where the injection site was (probably scar tissue again), but nothing like she was suffering previously. Her arm clearly has a large dip where so much muscle tissue was removed, but it doesn't notice now she's older (53), although she tends to wear short sleeves rather than tank tops. Sunburn irritates it was well. However in comparison with the pain she suffered for a year or so when the inflamed area was present, she considers the operation was a complete success. This was all back in 1982, and medical staff here were non-plussed by the injection reaction at the time. She didn't get any systemic problems she was aware of, just around the injection site (although that was so painful it may have masked other symptoms I suppose). It was quite a drastic cure but it worked, and of course she went on to marry a great guy and have wonderful children while still juggling a successful career etc....
My wife's comments :
"They didn't treat it with anything. The only other cases I came across then (as it was pre-internet) were either allergy to the aluminium used in preparing the inoculum or in the 3rd world women reacted to the short chains of an 'oil' in the adjuvant which normally used long-chain molecules but not the inoculum itself.
Whatever it was, it was as instant as the day after the injection I could not lift or touch my arm.
Mine flared up every 2 weeks eventually like clockwork.
The first local excision did work but because they cut it out when it was 'quiet' they could not discern where it was. I told them they had cut it in half but of course they didn't believe me. So when it re-appeared on schedule 2 weeks later they decided to cut a huge amount out just to be sure. If they had done the 2nd op first it would not have needed to be so drastic but still sizeable."
I hope this is of some help.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
} ------------------------------------------------------------------------ ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ---- } } Good afternoon, good evening, } dear listers, } } Allow to ask a question perhaps not usual for this forum: } } We do have a patient suffering from several pain due to a post-vaccination } problem (due to accumulation of Al-precipitates in intermuscular } macrophages) , namely } } macrophagic myofasciitis, } } luckily NOT systemic or generalized. } } If there is anybody out there who has knowledge about a specific and } reliable treatment for such a condition I greatly should appreciate your } comment. } (Unfortunately I was not able to localize any reference for } successful { } treatment). } } } If -perhaps- I have broken rules for using this listserver, please } apologize. It is just to seek help for a young patient. } } Thank you and } cordially yours } } Wolfgang Muss, PhD. } Salzburg, Austria }
==============================Original Headers============================== 16, 28 -- From keith.morris-at-ucl.ac.uk Wed Mar 1 04:17:37 2006 16, 28 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) 16, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21AHZEt004779 16, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 04:17:36 -0600 16, 28 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 16, 28 -- by vscani-d.ucl.ac.uk with smtp (Exim 4.51) 16, 28 -- id 1FEOOh-0007P3-Ns; Wed, 01 Mar 2006 10:17:31 +0000 16, 28 -- Message-ID: {003f01c63d19$3f7b26b0$7b865290-at-keithhigrade} 16, 28 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 16, 28 -- To: {Microscopy-at-microscopy.com} 16, 28 -- Cc: {W.Muss-at-salk.at} 16, 28 -- References: {200602271813.k1RID3wk015939-at-ns.microscopy.com} 16, 28 -- Subject: Re: [Microscopy] Vaccination complications: macrophagic myofasciitis 16, 28 -- Date: Wed, 1 Mar 2006 10:16:47 -0000 16, 28 -- MIME-Version: 1.0 16, 28 -- Content-Type: text/plain; 16, 28 -- format=flowed; 16, 28 -- charset="iso-8859-1"; 16, 28 -- reply-type=original 16, 28 -- Content-Transfer-Encoding: 7bit 16, 28 -- X-Priority: 3 16, 28 -- X-MSMail-Priority: Normal 16, 28 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 16, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 16, 28 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 16, 28 -- X-UCL-MailScanner: Found to be clean 16, 28 -- X-UCL-MailScanner-SpamCheck: 16, 28 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk ==============================End of - Headers==============================
==============================Original Headers============================== 31, 28 -- From W.Muss-at-salk.at Wed Mar 1 10:41:27 2006 31, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 31, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21GfQch014880 31, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 10:41:26 -0600 31, 28 -- Received: from localhost (localhost [127.0.0.1]) 31, 28 -- by hermes.lks.at (Postfix) with ESMTP id CF10C5A9020; 31, 28 -- Wed, 1 Mar 2006 17:41:23 +0100 (CET) 31, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 31, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 31, 28 -- with ESMTP id 60933-01; Wed, 1 Mar 2006 17:41:23 +0100 (CET) 31, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 31, 28 -- by hermes.lks.at (Postfix) with SMTP id 67DC15A900A; 31, 28 -- Wed, 1 Mar 2006 17:41:23 +0100 (CET) 31, 28 -- Received: by localhost with Microsoft MAPI; Wed, 1 Mar 2006 17:41:10 +0100 31, 28 -- Message-ID: {01C63D57.53DD50E0.W.Muss-at-salk.at} 31, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 31, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 31, 28 -- To: "'keith.morris-at-ucl.ac.uk'" {keith.morris-at-ucl.ac.uk} 31, 28 -- Cc: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 31, 28 -- Subject: [Microscopy] Re(Summ): Vaccination complications: macrophagic myofasciitis 31, 28 -- Date: Wed, 1 Mar 2006 17:41:09 +0100 31, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 31, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 31, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 31, 28 -- MIME-Version: 1.0 31, 28 -- Content-Type: text/plain; charset="us-ascii" 31, 28 -- Content-Transfer-Encoding: 7bit 31, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Dear Hong, perhaps this citation could be also of value for your studies: (indeed I too was not able to find some better technical solution than : silver sulphide reaction(s), Gorm Danscher's methods, and Silver enhancement as Rick Powell formulated in his recent mail.
All best wishes Wolfgang Muss Salzburg/Austria
copied from HISTONET-Listserver to be found at: http://www.histosearch.com/histonet/May01/RE.Manganesestaining.html RE: Manganese staining
Roy Ellis {roy.ellis-at-imvs.sa.gov.au}
Beth, The following reference to manganese is found in 'Theory and strategy in histochemistry' edited by Hans Lyon and published by Springer-Verlag. After treating a section with benzothiazolylazonaphthol, manganese deposits will stain blue. The method is not specific as other metals, namely cadmium, zinc and nickel also stain blue. Post-differentiation with 1% oxine (8-hydroxyquinoline) in 75% ethanol turned tissue cadmium from blue to red but zinc remained blue. Regards Roy Ellis mailto:roy.ellis-at-imvs.sa.gov.au }
-----Original Message----- } From: Histo-Scientific Research Laboratories } [mailto:histosci-at-shentel.net] } Sent: Thursday, 3 May 2001 21:38 } To: Linda McGraf } Subject: Manganese staining } } } Dear Histonetters, } } Has anyone ever heard of staining for localization of manganese in animal } tissue? We have looked through our staining books and have come up with } nothing. If anyone has ever heard of such a procedure, please share any } info you may have. } } Thank you, } } Beth Poole } HSRL } beth-at-hsrl.org } (540)459-8211 } fax: (540)459-8217 } www.hsrl.org } 137 South Main Street } Woodstock, VA 22664
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America We have a PI here wants to localize manganese in tissue. Does anyone know a way of doing this without using X-ray microanalysis? Thank you.
Hong Emory
==============================Original Headers============================== 4, 16 -- From hyi-at-emory.edu Tue Feb 28 15:56:35 2006 4, 16 -- Received: from sccrmhc14.comcast.net (sccrmhc14.comcast.net [63.240.77.84]) 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1SLuZKn032163 4, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 15:56:35 -0600 4, 16 -- Received: from [24.30.86.130] (c-24-30-86-130.hsd1.ga.comcast.net[24.30.86.130]) 4, 16 -- by comcast.net (sccrmhc14) with SMTP 4, 16 -- id {2006022821563401400ebg66e} ; Tue, 28 Feb 2006 21:56:34 +0000 4, 16 -- Mime-Version: 1.0 (Apple Message framework v622) 4, 16 -- Content-Transfer-Encoding: 7bit 4, 16 -- Message-Id: {e81b3f049cea790cdb8341be7e80af1a-at-emory.edu} 4, 16 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 16 -- To: Microscopy-at-microscopy.com 4, 16 -- From: Hong Yi {hyi-at-emory.edu} 4, 16 -- Subject: Manganese 4, 16 -- Date: Tue, 28 Feb 2006 16:56:38 -0500 4, 16 -- X-Mailer: Apple Mail (2.622) ==============================End of - Headers==============================
==============================Original Headers============================== 15, 28 -- From W.Muss-at-salk.at Wed Mar 1 10:45:31 2006 15, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 15, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21GjSok015853 15, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 10:45:29 -0600 15, 28 -- Received: from localhost (localhost [127.0.0.1]) 15, 28 -- by hermes.lks.at (Postfix) with ESMTP id 5EE5C5A9044; 15, 28 -- Wed, 1 Mar 2006 17:45:23 +0100 (CET) 15, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 15, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 15, 28 -- with ESMTP id 61136-01; Wed, 1 Mar 2006 17:45:22 +0100 (CET) 15, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 15, 28 -- by hermes.lks.at (Postfix) with SMTP id EE1B65A9041; 15, 28 -- Wed, 1 Mar 2006 17:45:22 +0100 (CET) 15, 28 -- Received: by localhost with Microsoft MAPI; Wed, 1 Mar 2006 17:45:10 +0100 15, 28 -- Message-ID: {01C63D57.E2BB1B80.W.Muss-at-salk.at} 15, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 15, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 15, 28 -- To: "'hyi-at-emory.edu'" {hyi-at-emory.edu} 15, 28 -- Cc: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 15, 28 -- Subject: AW: [Microscopy] Manganese 15, 28 -- Date: Wed, 1 Mar 2006 17:45:09 +0100 15, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 15, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 15, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 15, 28 -- MIME-Version: 1.0 15, 28 -- Content-Type: text/plain; charset="us-ascii" 15, 28 -- Content-Transfer-Encoding: 7bit 15, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Cr is known to oxidize quickly so I eliminated that one.
C would be good but the specimens may contain C.
I failed to list the elements I might see: B, C, O, F, Na, Mg, P, Si, Al, S, Cl, Ar, K, Ca, Fe, Co, Mo, In, Cu, Hf, W, Ga, As. Not all at once!
It is mostly the lower Zs that are a problem with low energy peaks around 2KeV.
Perhaps Pd alone is a good choice? This does not occur in organic dielectrics or other specimens.
gary g.
At 08:10 AM 3/1/2006, you wrote:
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==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Wed Mar 1 11:22:41 2006 13, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k21HMdss025692 13, 20 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 11:22:40 -0600 13, 20 -- Received: (qmail 21705 invoked from network); 1 Mar 2006 09:22:39 -0800 13, 20 -- Received: by simscan 1.1.0 ppid: 21702, pid: 21703, t: 0.1877s 13, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 20 -- by qsmtp3 with SMTP; 1 Mar 2006 09:22:39 -0800 13, 20 -- Message-Id: {6.2.3.4.2.20060301091618.0205cf68-at-mail.calweb.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 13, 20 -- Date: Wed, 01 Mar 2006 09:22:39 -0800 13, 20 -- To: ahlst007-at-umn.edu 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] RE: Low Z peak pileup] 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200603011610.k21GAN67009038-at-ns.microscopy.com} 13, 20 -- References: {200603011610.k21GAN67009038-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
How about TEM EELS? Does anyone out there know if this will work?
Aloha, Tina
_____________________________________________________________________________ Dear All: We have a PI here wants to localize manganese in tissue. Does anyone know a way of doing this without using X-ray microanalysis? Thank you.
Hong Emory
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * Biological Electron Microscope Facility * (808) 956-6251 * University of Hawaii at Manoa * * http://www.pbrc.hawaii.edu/bemf ****************************************************************************
==============================Original Headers============================== 8, 19 -- From tina-at-pbrc.hawaii.edu Wed Mar 1 12:52:51 2006 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 8, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21IqmNf013586 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 12:52:50 -0600 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k21Iqh0B022445 8, 19 -- (version=TLSv1/SSLv3 cipher=AES256-SHA bits=256 verify=NO) 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 08:52:44 -1000 (HST) 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k21IqgtI022442 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 08:52:43 -1000 (HST) 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 8, 19 -- Date: Wed, 1 Mar 2006 08:52:41 -1000 (HST) 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 8, 19 -- X-Sender: tina-at-halia 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 8, 19 -- Subject: AW: Manganese 8, 19 -- Message-ID: {Pine.GSO.4.21.0603010849240.22388-100000-at-halia} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
I have a student making C-Pt replicas. He shadows with Pt at 45 degree tilt with rotation. The protocol he has found then depoists C at 90 degrees to the source with rotation. He wants to know why he needs to change the angle of tilt to do the carbon. I could not really give him a good explanation. Can any of you help?
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21Jx6ko024033 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 13:59:07 -0600 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 4, 22 -- (authenticated bits=0) 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k21Jx5pL2826334 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 14:59:05 -0500 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 4, 22 -- X-Accept-Language: en-us, en 4, 22 -- MIME-Version: 1.0 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Replicas 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
When doing conventional replicas (static specimens, no rotation) the Pt evaporation is conducted at an angle to generate the shadows. The carbon is then evaporated at 90 degrees (directly above the specimen) to fill in the voids or gaps (shadows) generated during the Pt evaporation. This strengthens the replica.
In your case, you are rotating the specimen on a turntable in both cases. Nonetheless, even though the Pt evaporation is being carried out with rotation, you will still have some gaps (otherwise you would not see any shadows). The carbon (since it is being evaporated directly overhead) will not land in the same areas as the Pt but will more uniformly coat the specimen and fill in the shadows (gaps). Although it fills in the gaps, its low density does not interfere with the shadows generated by the Pt.
So, the reason for the different angles is to make the replica stronger by filling in any voids or gaps.
JB
} I have a student making C-Pt replicas. He shadows with Pt at 45 degree } tilt with rotation. The protocol he has found then depoists C at 90 } degrees to the source with rotation. He wants to know why he needs to } change the angle of tilt to do the carbon. I could not really give him } a good explanation. Can any of you help? } } Greg } } -- } Gregory W. Erdos, Ph.D, } Assistant Director, Biotechnology Program } Scientific Director, EM Core Lab } P.O. Box 118525 } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } Phone: 352-392-1295 } Fax: 352-846-0251 } } } ==============================Original Headers============================== } 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 } 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) } 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k21Jx6ko024033 } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } 13:59:07 -0600 } 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu } [10.227.60.63]) } 4, 22 -- (authenticated bits=0) } 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id } k21Jx5pL2826334 } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } 14:59:05 -0500 } 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} } 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 } 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} } 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) } 4, 22 -- X-Accept-Language: en-us, en } 4, 22 -- MIME-Version: 1.0 } 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" } {Microscopy-at-MSA.Microscopy.Com} } 4, 22 -- Subject: Replicas } 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 22 -- Content-Transfer-Encoding: 7bit } 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } 4, 22 -- X-Scanned-By: CNS Open Systems Group } (http://open-systems.ufl.edu/services/smtp-relay/) } 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group } (http://open-systems.ufl.edu/services/smtp-relay/) } ==============================End of - Headers==============================
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==============================Original Headers============================== 9, 18 -- From bozzola-at-siu.edu Wed Mar 1 14:28:03 2006 9, 18 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21KS3TZ029200 9, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 14:28:03 -0600 9, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 9, 18 -- by abbmta1.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k21KS1FY023612 9, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 14:28:02 -0600 (CST) 9, 18 -- Mime-Version: 1.0 9, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 9, 18 -- Message-Id: {p06110407c02bb21f9f99-at-[131.230.177.142]} 9, 18 -- In-Reply-To: {200603012010.k21KAXCC026054-at-ns.microscopy.com} 9, 18 -- References: {200603012010.k21KAXCC026054-at-ns.microscopy.com} 9, 18 -- Date: Wed, 1 Mar 2006 14:28:00 -0600 9, 18 -- To: Microscopy-at-msa.microscopy.com 9, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 9, 18 -- Subject: Re: [Microscopy] Replicas 9, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 9, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
We used to use Pt/C replicas to show surface relief. We would shadow a cellulose acetate surface replica with Pt/C at a small angle to the surface to highlight the surface texture, then deposit C normal to the surface to provide the support film. After the C film deposition the film was removed from the replica surface and mounted on a copper grid.
It is still useful when you want to see the height of features on a surface. If you know the shadow angle, you can easily calculate the height. I am not aware of doing sample rotation during the shadowing step, but then I'm a materials guy and am not familiar with some of the bio techniques.
Good luck, Henk
At 03:16 PM 03/01/06, gwe-at-ufl.edu wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
==============================Original Headers============================== 10, 26 -- From colijn.1-at-osu.edu Wed Mar 1 14:54:52 2006 10, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21KspXt003731 10, 26 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 14:54:51 -0600 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 10, 26 -- id {01LZJO8BC9XS9V57H2-at-er6s1.eng.ohio-state.edu} for 10, 26 -- microscopy-at-microscopy.com; Wed, 01 Mar 2006 15:54:50 -0500 (EST) 10, 26 -- Received: from HOC1.ecr6.ohio-state.edu 10, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 10, 26 -- (PMDF V6.2-1x5 #31056) 10, 26 -- with ESMTPA id {01LZJO8AF6HG9VA6YH-at-er6s1.eng.ohio-state.edu} ; Wed, 10, 26 -- 01 Mar 2006 15:54:50 -0500 (EST) 10, 26 -- Date: Wed, 01 Mar 2006 15:54:47 -0500 10, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: [Microscopy] Replicas 10, 26 -- In-reply-to: {200603012016.k21KGm5p027267-at-ns.microscopy.com} 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- X-Sender: colijn-at-mail.ecr6.ohio-state.edu 10, 26 -- To: gwe-at-ufl.edu, Microscopy Listserver {microscopy-at-microscopy.com} 10, 26 -- Message-id: {6.1.0.6.2.20060301154803.02ee3940-at-mail.ecr6.ohio-state.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.0.6 10, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: {200603012016.k21KGm5p027267-at-ns.microscopy.com} ==============================End of - Headers==============================
Unless you change angles, the Pt and C will land in the same places. By changing the angle from 45 to 90 deg, the carbon covers pretty much everything (no shadows would be generated) since it comes straight down.
An interesting demonstration would be to mount an object on a turntable at the appropriate angles and use some colored spray paints.
} If the stage is tilted and rotating would the carbon not be able to } get in all the gaps? } } bozzola-at-siu.edu wrote: } } } } When doing conventional replicas (static specimens, no rotation) } } the Pt evaporation is conducted at an angle to generate the } } shadows. The carbon is then evaporated at 90 degrees (directly } } above the specimen) to fill in the voids or gaps (shadows) } } generated during the Pt evaporation. This strengthens the replica. } } } } In your case, you are rotating the specimen on a turntable in both } } cases. Nonetheless, even though the Pt evaporation is being carried } } out with rotation, you will still have some gaps (otherwise you } } would not see any shadows). The carbon (since it is being } } evaporated directly overhead) will not land in the same areas as } } the Pt but will more uniformly coat the specimen and fill in the } } shadows (gaps). Although it fills in the gaps, its low density does } } not interfere with the shadows generated by the Pt. } } } } So, the reason for the different angles is to make the replica } } stronger by filling in any voids or gaps. } } } } JB } } } } } I have a student making C-Pt replicas. He shadows with Pt at 45 degree } } } tilt with rotation. The protocol he has found then depoists C at 90 } } } degrees to the source with rotation. He wants to know why he needs to } } } change the angle of tilt to do the carbon. I could not really give him } } } a good explanation. Can any of you help? } } } } } } Greg } } } } } } -- } } } Gregory W. Erdos, Ph.D, } } } Assistant Director, Biotechnology Program } } } Scientific Director, EM Core Lab } } } P.O. Box 118525 } } } University of Florida } } } Gainesville, FL 32611 } } } gwe-at-ufl.edu } } } Phone: 352-392-1295 } } } Fax: 352-846-0251 } } } } } } } } } ==============================Original Headers============================== } } } 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 } } } 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) } } } 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } } k21Jx6ko024033 } } } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } } } 13:59:07 -0600 } } } 4, 22 -- Received: from [10.227.60.63] } } } (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) } } } 4, 22 -- (authenticated bits=0) } } } 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id } } } k21Jx5pL2826334 } } } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } } } 14:59:05 -0500 } } } 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} } } } 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 } } } 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} } } } 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) } } } 4, 22 -- X-Accept-Language: en-us, en } } } 4, 22 -- MIME-Version: 1.0 } } } 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" } } } {Microscopy-at-MSA.Microscopy.Com} } } } 4, 22 -- Subject: Replicas } } } 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } } 4, 22 -- Content-Transfer-Encoding: 7bit } } } 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } } } 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } } } 4, 22 -- X-Scanned-By: CNS Open Systems Group } } } (http://open-systems.ufl.edu/services/smtp-relay/) } } } 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group } } } (http://open-systems.ufl.edu/services/smtp-relay/) } } } ==============================End of - Headers============================== } } } } } } } } } } } } } } -- } Gregory W. Erdos, Ph.D, } Assistant Director, Biotechnology Program } Scientific Director, EM Core Lab } P.O. Box 118525 } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } Phone: 352-392-1295 } Fax: 352-846-0251
==============================Original Headers============================== 7, 19 -- From bozzola-at-siu.edu Wed Mar 1 15:27:29 2006 7, 19 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 7, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21LRRIn013166 7, 19 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 15:27:28 -0600 7, 19 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 7, 19 -- by abbmta1.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k21LROtr015034 7, 19 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 15:27:25 -0600 (CST) 7, 19 -- Mime-Version: 1.0 7, 19 -- X-Sender: bozzola-at-saluki-mail.siu.edu 7, 19 -- Message-Id: {p06110409c02bc11220b3-at-[131.230.177.142]} 7, 19 -- In-Reply-To: {44060A15.9080709-at-ufl.edu} 7, 19 -- References: {200603012037.k21KbwL4031717-at-ns.microscopy.com} 7, 19 -- {44060A15.9080709-at-ufl.edu} 7, 19 -- Date: Wed, 1 Mar 2006 15:27:23 -0600 7, 19 -- To: Microscopy-at-msa.microscopy.com 7, 19 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 7, 19 -- Subject: Re: [Microscopy] Re: Replicas 7, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 19 -- X-MASF: 0.00% ==============================End of - Headers==============================
Bozzola and Russell do a good job at diagrammatically showing why (if you want pictures - and well JB does a quick service to the list as well).
The Pt doesn't make a continuous film, even with rotation- esp if your samples have significant surface relief.
At 90° the carbon can be added uniformly and in a topographically neutral manner. Think about the carbon as the support for the Pt.
At least that's how I thought of the process. I had a luxury of having two sources: A point target with the Pt and a separate one for the carbon. And with the 90 degrees, I'd say drop the rotation maybe.
Does that help? When explaining something like this I always resort to drawing pictures.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: gwe-at-ufl.edu [mailto:gwe-at-ufl.edu] Sent: Wednesday, March 01, 2006 3:47 PM To: Williams, Geoffrey
I have a student making C-Pt replicas. He shadows with Pt at 45 degree tilt with rotation. The protocol he has found then depoists C at 90 degrees to the source with rotation. He wants to know why he needs to change the angle of tilt to do the carbon. I could not really give him a good explanation. Can any of you help?
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21Jx6ko024033 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 13:59:07 -0600 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 4, 22 -- (authenticated bits=0) 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k21Jx5pL2826334 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 14:59:05 -0500 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 4, 22 -- X-Accept-Language: en-us, en 4, 22 -- MIME-Version: 1.0 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Replicas 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
==============================Original Headers============================== 16, 29 -- From Geoffrey_Williams-at-brown.edu Wed Mar 1 15:40:49 2006 16, 29 -- Received: from scorpio.services.brown.edu (scorpio.services.brown.edu [128.148.106.155]) 16, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21Lel6f017722 16, 29 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 15:40:48 -0600 16, 29 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 16, 29 -- by scorpio.services.brown.edu (Switch-3.1.7/Switch-3.1.7/) with ESMTP id k21LekQ1020973; 16, 29 -- Wed, 1 Mar 2006 16:40:47 -0500 (EST) 16, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 16, 29 -- Wed, 1 Mar 2006 16:40:46 -0500 16, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 16, 29 -- Content-class: urn:content-classes:message 16, 29 -- MIME-Version: 1.0 16, 29 -- Content-Type: text/plain; 16, 29 -- charset="iso-8859-1" 16, 29 -- Subject: RE: [Microscopy] Replicas 16, 29 -- Date: Wed, 1 Mar 2006 16:40:45 -0500 16, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F042303E5-at-MAIL1.AD.Brown.Edu} 16, 29 -- X-MS-Has-Attach: 16, 29 -- X-MS-TNEF-Correlator: 16, 29 -- Thread-Topic: [Microscopy] Replicas 16, 29 -- Thread-Index: AcY9cVbxc5ZMZm11QeSykRvDszMUyAABqVxg 16, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 16, 29 -- To: {gwe-at-ufl.edu} , {Microscopy-at-microscopy.com} 16, 29 -- X-OriginalArrivalTime: 01 Mar 2006 21:40:46.0700 (UTC) FILETIME=[CCBDD2C0:01C63D78] 16, 29 -- X-Brown-Proofpoint: Not Infected 16, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06021401 definitions=3.0.0-06030104 16, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06021401 definitions=3.0.0-06030104 16, 29 -- Content-Transfer-Encoding: 8bit 16, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k21Lel6f017722 ==============================End of - Headers==============================
I believe that they went with Pt/C to form small clusters for better resolution. It think that pure metals will nucleate in larger islands.
Henk
At 04:46 PM 03/01/06, you wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
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Email: mccaulak-at-wfu.edu Name: Anita McCauley
Organization: Wake Forest University
Title-Subject: [Filtered] fluorolume illuminator
Question: A colleague at a neighoring institution recently found what she described as a "fluorolume, old-style fluorescent illuminator". She asked me if I knew anything about how to operate it. I have never heard of a fluorolume and so am unable to help her right now. I would appreciate any responses regarding what it is, how it works, issues associated with it's use, etc...
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Email: cbucana-at-mdanderson.org Name: C Bucana
Organization: UT MD. Anderson Cancer Center
Title-Subject: [Filtered] SEM of bat embryos
Question: I would appreciate any information on processing bat embryos for scanning electron microscopy. We were given embryos at different stages of development and we fixed them in glutaraldehyde/paraformaldehyde fixative, fixed in osmium tetroxide, dehydrated in increasing concentration of ethanol and dehydrated in HMDS before air drying and metal coating. Upon examination under the dissecting microscope, we found that the skin has separated from the rest of the embryo, i.e. the skin looks like it has balooned or it did not shrink while the rest of the embryo shrank. Any suggestions or comments will be greatly appreciated.
This Question was submitted to Ask-A-Microscopist by (exploratorium-at-tiscali.it) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, March 1, 2006 at 17:02:14 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both exploratorium-at-tiscali.it as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: exploratorium-at-tiscali.it Name: giovanni de caro
Organization: associazione luigi montalbÚ
Education: 9-12th Grade High School
Location: Campobasso, Italy
Title: SEM operation video
Question: I am looking for a video (VHS or DVD) dealing with the practical operation of an older SEM. We are going to restart and operate an AMR 1000 SEM in our natural science museum for youngsters in Italy (http://web.tiscali.it/exploratorium). Of course I do not expect a documentary ilustrating THIS specific instrument, I would be more than happy of something showing the operation of this calss of instruments. I also would like to have a video on SEM specimen preparation. Thank you for your kind help.
This Question was submitted to Ask-A-Microscopist by (l_mtl-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, February 28, 2006 at 08:14:52 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both l_mtl-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: l_mtl-at-yahoo.com Name: Reza
Organization: TEHRAN UNI
Education: Undergraduate College
Location: Tehran,iran
Title: Magnification marker
Question: I have some TEM pictured that i got from scanned negative. these pictures have no magnification marker. my question is that how i can put a magnification marker on pictures when i have only negative and camrea length? thanks in advance
With an absorption edge ca 650 eV, Mn has a weak signal for EELS,. But it is doable, especially if the concentration is high enough. With freeze substituted cyanobacteria we get a cytosolic distribution of Mn in EELS ESI images.
Howard
On Mar 1, 2006, at 1:11 PM, tina-at-pbrc.hawaii.edu wrote:
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R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility Danforth Plant Science Center 975 N. Warson Rd. St. Louis, MO 63132
ph 314-587-1261 fx 314-587-1361 cell 314-378-2409 rhberg-at-danforthcenter.org www.danforthcenter.org visit this educational resource: http://www.danforthcenter.org/Cells/
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I want to provide a different perspective on the use of replicas. High-resolution Pt/C shadowing and replication provided important insights into the size and shape of polymers beginning over 40 years ago. The first images of DNA molecules were made this way. However, in my opinion, this methodology has largely been supplanted by the use of Atomic Force Microscopy, both for direct height measurements (available in Pt/C replicas by measuring shadow lengths when the coating is deposited from one direction only) and for imaging molecular contours.
As an everyday example, consider that making a magnetic read and write head for a hard disk drive requires controlling the relative heights of several different regions to a tolerance of ca. 1 nm. AFM supports production by providing a rapid means of offline analysis, far faster and more precise than any replica method could be. In the biopolymer area, for more than 10 years, it has been relatively easy to prepare dispersions of molecules on smooth surfaces like mica for AFM imaging. We have done some of this work ourselves, and examples can be seen at:
Disclaimer: ASM provides analytical services using AFM and I benefit from increasing the demand for AFM data.
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
----- Original Message ----- From: gwe-at-ufl.edu To: donc-at-asmicro.com Sent: Wednesday, March 01, 2006 3:37 PM Subject: [a] [Microscopy] Replicas
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I have a student making C-Pt replicas. He shadows with Pt at 45 degree tilt with rotation. The protocol he has found then depoists C at 90 degrees to the source with rotation. He wants to know why he needs to change the angle of tilt to do the carbon. I could not really give him a good explanation. Can any of you help?
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
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==============================Original Headers============================== 24, 22 -- From donc-at-asmicro.com Wed Mar 1 18:42:03 2006 24, 22 -- Received: from smtp103.sbc.mail.re2.yahoo.com (smtp103.sbc.mail.re2.yahoo.com [68.142.229.102]) 24, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k220g1aL011199 24, 22 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 18:42:02 -0600 24, 22 -- Received: (qmail 96736 invoked from network); 2 Mar 2006 00:42:01 -0000 24, 22 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.57.205.80 with login) 24, 22 -- by smtp103.sbc.mail.re2.yahoo.com with SMTP; 2 Mar 2006 00:42:00 -0000 24, 22 -- Message-ID: {000001c63d91$beaf7580$c901a8c0-at-ASM11} 24, 22 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 24, 22 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 24, 22 -- To: {gwe-at-ufl.edu} , "Microscopy List" {microscopy-at-microscopy.com} 24, 22 -- References: {200603012037.k21Kb5TY031477-at-ns.microscopy.com} 24, 22 -- Subject: Re: [a] [Microscopy] Replicas 24, 22 -- Date: Wed, 1 Mar 2006 18:06:00 -0500 24, 22 -- MIME-Version: 1.0 24, 22 -- Content-Type: text/plain; 24, 22 -- charset="iso-8859-1" 24, 22 -- Content-Transfer-Encoding: 7bit 24, 22 -- X-Priority: 3 24, 22 -- X-MSMail-Priority: Normal 24, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 24, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
One of my users has problems getting a good grid with negatively stained viral particles. She floats the grid on the virus+stain droplet, picks up the grid and then dries by touching against filter paper. It sounds like a pretty standard procedure but she often found that the support film ("store-bought" carbon coated formvar on either 300 or 400 mesh copper grid) is broken after the procedure. And occasionally, almost every hole is torn. Is there any tricks to prevent this?
I have experienced similar broken film but it was in formvar coated slot grids. After picking up a group of ribbons, the support film broke when the grid is dried. I always thought that I was just careless during the handling causing the film to crack and eventually the surface tension tore the film completely. However, that would be unlikely for the 300 or 400 mesh grids which are supported by so many grid bars? Thanks for any advice.
-- Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519) The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
==============================Original Headers============================== 4, 19 -- From wpchan-at-u.washington.edu Wed Mar 1 19:57:44 2006 4, 19 -- Received: from mxout5.cac.washington.edu (mxout5.cac.washington.edu [140.142.32.135]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k221vgwE007728 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 19:57:43 -0600 4, 19 -- Received: from homer23.u.washington.edu (homer23.u.washington.edu [140.142.12.141]) 4, 19 -- by mxout5.cac.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k221vfws006724 4, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 17:57:42 -0800 4, 19 -- Received: from localhost (wpchan-at-localhost) 4, 19 -- by homer23.u.washington.edu (8.13.5+UW05.10/8.13.5+Submit) with ESMTP id k221vf6I025767 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 17:57:41 -0800 4, 19 -- Date: Wed, 1 Mar 2006 17:57:41 -0800 (PST) 4, 19 -- From: "W. Chan" {wpchan-at-u.washington.edu} 4, 19 -- To: microscopy-at-microscopy.com 4, 19 -- Subject: viral particles 4, 19 -- Message-ID: {Pine.LNX.4.64.0603011722450.24739-at-homer23.u.washington.edu} 4, 19 -- MIME-Version: 1.0 4, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 19 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
The previous responders are correct in that the carbon makes the support film and the heavy metal creates shadows of the surface structure. The heavy metal angle is really a variable and should not always be set at 45 degrees per some protocol. The shallower the surface structure, the lower the Pt dep angle. As was stated a picture here would be a big help, but think of a 100nm high step vs a 2 nm high step on an otherwise smooth substrate. From my experience 45 degrees would be OK to shadow the 100nm step into visibility (actually a bit high). A Pt dep angle of 10 to 15 degrees would be better for the 2 nm step--longer shadows. Whatever angle you use you can make a rough calculation of step height from the geometry of the Pt shadowing--assuming the replica stays flat vs. assuming a potato chip shape.
Aside from AFM imaging, a Pt-C replica will give better topographic resolution of extremely small steps on a flat surface where secondary electron SEM contrast is weak, IMHO better than a super-duper SEM . Single-atom high growth steps on a growing crystal surface for example.
There is another "lost art" replication method I would love to see someone perform and send me the images to display in Microscopy Today: (I no longer have access to an e.m. lab). Pull a carbon replica of a fairly rough surface. Do not apply a heavy metal shadow. Put the naked carbon film into a TEM at 100keV or so. Tilt the specimen as high as your goniometer stage allows--45 to 60 degrees is best. Image with a small objective aperture in the bright-field position allowing the unscattered main beam to pass. Find an interesting step or structure using the weak contrast available in this mode. Then slightly displace the objective aperture so that the bright part of the main beam is *just* outside the aperture (or tilt the beam leaving the aperture centered to obtain the same effect). In other words, you are making a dark-field image using the "tails" of the main beam. Refocus. The result should be a sharp, high contrast, positive image that looks like and has the resolution of a high quality SEM image.
Ron Anderson, Editor Microscopy Today
gwe-at-ufl.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a student making C-Pt replicas. He shadows with Pt at 45 degree } tilt with rotation. The protocol he has found then depoists C at 90 } degrees to the source with rotation. He wants to know why he needs to } change the angle of tilt to do the carbon. I could not really give him } a good explanation. Can any of you help? } } Greg } }
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unfortunately I do not have any idea how to be of help with handling such an illumination system.....but my personal interest in the request lead to the following search results:
............... ".....The basic tungsten source was the AO Ortho-illuminator and the high pressure mercury arc source was the AO Fluorolume illuminator equipped with a Corning (filter..)....." Source: Interchangeable Tungsten Filament and Mercury Arc Illuminator Adaptation for the Interference Microscope E. W. Lowrance Transactions of the American Microscopical Society, Vol. 86, No. 2 (Apr., 1967) , pp. 223-224 This journal is licensed to JSTOR by American Microscopical Society ==} http://links.jstor.org/sici?sici=0003-0023(196704)86%3A2%3C223%3AITFAMA% 3E2.0.CO%3B2-W
Structural Changes During Bean Leaf Abscission Peter C. Scott, Lillian W. Miller, Barbara D. Webster, A. C. Leopold American Journal of Botany, Vol. 54, No. 6 (Jul., 1967) , pp. 730-734 This journal is licensed to JSTOR by American Microscopical Society
Other Citations: Reference: Cellular and Molecular Life Sciences (CMLS) Publisher: Birkhauser Basel ISSN: 1420-682X (Paper) 1420-9071 (Online) DOI: 10.1007/BF01985701 Issue: Volume 37, Number 8
Question: A colleague at a neighoring institution recently found what she described as a "fluorolume, old-style fluorescent illuminator". She asked me if I knew anything about how to operate it. I have never heard of a fluorolume and so am unable to help her right now. I would appreciate any responses regarding what it is, how it works, issues associated with it's use, etc...
I have usually found that coatings collapse when the virus still has a lot of extraneous material associated with it such as from faecal samples or a not very pure cell pellet. I assumed it was mainly a heating and charging effect because of the high levels of background organic material which will swamp the conducting capacity of the carbon and copper on the grid. You never normally see it on pure isolates such as T4 phage. The simplest answer might be to dilute the drop until the grid stops breaking or spin out as much of the background as possible.
Your slot grid problem can easily happen because of flexing of the grid but it may also be weakened if you just coat the flat slot grids with plastic. I usually bend them slightly upwards so the plastic stretched across the and slot can't be damaged when it dries out.
I apologise if you have thought of all of this, already.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: wpchan-at-u.washington.edu
Dear Reza,
as I understand your question, in my opinion it is not possible to "put a magnification marker on pictures when (...you....) have only negative and camera length".......yes, you COULD DO that....but without guarantee for a "real" magnification reference.
2 solutions (only one really will work properly):
Solution 1: "real magnification" You must know the original magnification of the (T)EM-system onto the "negative".... (i.e.: primary magnification of EM-system at the correct kV-setting [= 50, 80, 100 kV; normally a certification test of magnification should be included in delivery papers of an instrument and should not change until major repair like lifting column parts, altering major adjustments of lenses etc.] times camera factor [depending on camera system you use [ e.g. 35 mm different will be a camera factor compared to other imaging/negative formats] = primary magnification of structures ON the NEGATIVE.
OR, right from the beginning, you MUST KNOW the negative's magnification.
If you then enlarge (secondary magnification) by means of an ancillary enlarger system onto positive paper, you have to multiply primary magnification ON the NEGATIVE with the "factor" of your (secondary) enlarger....on a positive you then can draw bars with the apropriate length due to secondary enlargement, scanning the positive, you will have included the "real" and correct magnification of structure/image....
Solution two: this only will result in a very "approximatively" set magnification bar: if there is included a sructure of known "normal" length, width, diameter...etc you perhaps CAN DRAW a line indicating } ~ ?m { or so........
Real magnification out of an unknown magnified negative } times { known secondary enlargement (?camera length?) unfortunately results again in } unknown magnification {....
So at least you should search for the (primary) magnification of the negative......
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Title: Magnification marker
Question: I have some TEM pictured that i got from scanned negative. these pictures have no magnification marker. my question is that how i can put a magnification marker on pictures when i have only negative and camrea length? thanks in advance
For those of you who were curious as to why we were using rotary shadowing, I would refer you to the web page of Gary Borisy http://www.borisylab.northwestern.edu/pages/protocols/electmicrosc.html#metcoat
Look at Figs. 4,5,6,8 to see the result of such shadowing on cytoskeletal proteins.
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 5, 22 -- From gwe-at-ufl.edu Thu Mar 2 07:44:10 2006 5, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22Di9UC021977 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 2 Mar 2006 07:44:09 -0600 5, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 5, 22 -- (authenticated bits=0) 5, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k22Di6iY4321324 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 2 Mar 2006 08:44:07 -0500 5, 22 -- Message-ID: {4406F6A8.9090601-at-ufl.edu} 5, 22 -- Date: Thu, 02 Mar 2006 08:44:08 -0500 5, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 5, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 5, 22 -- X-Accept-Language: en-us, en 5, 22 -- MIME-Version: 1.0 5, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- Subject: Replicas FYI 5, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 5, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 5, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 5, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Pang, When you say that she dries the grids by touching them to filter paper....is she actually touching the face of the grids, or wicking the excess fluid by touching the edge of the grid to the paper? This is a critical difference. Its all in the interpretation of a written protocol. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 21 -- From lcgould-at-med.cornell.edu Thu Mar 2 08:06:57 2006 1, 21 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22E6vlu026341 1, 21 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 08:06:57 -0600 1, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 21 -- by smtp-gw2.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k22E6s4U023189 1, 21 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 09:06:54 -0500 (EST) 1, 21 -- Received: from [140.251.48.23] by mpx1.med.cornell.edu 1, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 21 -- with ESMTP id {0IVI003IA77HES40-at-mpx1.med.cornell.edu} for 1, 21 -- microscopy-at-microscopy.com; Thu, 02 Mar 2006 09:06:54 -0500 (EST) 1, 21 -- Date: Thu, 02 Mar 2006 09:02:51 -0500 1, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 21 -- Subject: Re: [Microscopy] viral particles 1, 21 -- In-reply-to: {200603020217.k222H7BQ014221-at-ns.microscopy.com} 1, 21 -- To: wpchan-at-u.washington.edu, Microscopy Listserver {microscopy-at-microscopy.com} 1, 21 -- Message-id: {p06200700c02cab041a7d-at-[140.251.48.23]} 1, 21 -- MIME-version: 1.0 1, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 21 -- References: {200603020217.k222H7BQ014221-at-ns.microscopy.com} 1, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.03.02.052605 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both edmarti-at-ceride.gov.ar as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] FET specification (Pre-amplifier 183 A)
Question: Dear all,
I'm working with a FRX Philips PV 9500 with a detector EDAX (Pre-amplifier 183 A). The beryllium window was replaced since it was broken, but the detector still doesn¥t work. After re-checking, I can conclude that the FET of the detector is broken (doesn't work). Then I need to know the technical specifications or data sheet of this FET.
Any help about specification would be appreciated.
Thanks,
TÈc. Ppal Elbio MartÌnez SECEGRIN - CERIDE - CONICET G¸emes 3450 3000 - Santa Fe Argentina email: edmarti-at-ceride.gov.ar
Instead of floating the filmed grid on the virus/stain mix try attaching the grid to a grid stick that has adhesive applied. Or you can put a piece of double-sided scotch tape onto the edge of a glass microscope slide and attach the extreme edge of the grid to that.
Now apply the virus/stain drop and after the appropriate time remove the liquid by touching the edge of the grid with a piece of filter paper.
This provides much gentler handling of the support film than lifting the grid off a droplet of solution where the surface tension creates quite a pull on the film as the grid is lifted away.
Certainly, using this technique, the support film should not rupture even on a 200 mesh grid.
Good luck,
Ted Dunn
--- wpchan-at-u.washington.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi } } One of my users has problems getting a good grid } with negatively stained } viral particles. She floats the grid on the } virus+stain droplet, picks up } the grid and then dries by touching against filter } paper. It sounds like } a pretty standard procedure but she often found that } the support film } ("store-bought" carbon coated formvar on either 300 } or 400 mesh copper } grid) is broken after the procedure. And } occasionally, almost every hole } is torn. Is there any tricks to prevent this? } } I have experienced similar broken film but it was in } formvar coated slot } grids. After picking up a group of ribbons, the } support film broke when } the grid is dried. I always thought that I was just } careless during the } handling causing the film to crack and eventually } the surface tension tore } the film completely. However, that would be } unlikely for the 300 or 400 } mesh grids which are supported by so many grid bars? } Thanks for any } advice. } } -- } Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB } A087, 206-685-1519) } The Biology Imaging Facility } (http://staff.washington.edu/wpchan/if/) } } ==============================Original } Headers============================== } 4, 19 -- From wpchan-at-u.washington.edu Wed Mar 1 } 19:57:44 2006 } 4, 19 -- Received: from mxout5.cac.washington.edu } (mxout5.cac.washington.edu [140.142.32.135]) } 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k221vgwE007728 } 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 } Mar 2006 19:57:43 -0600 } 4, 19 -- Received: from homer23.u.washington.edu } (homer23.u.washington.edu [140.142.12.141]) } 4, 19 -- by mxout5.cac.washington.edu } (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id } k221vfws006724 } 4, 19 -- (version=TLSv1/SSLv3 } cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 } Mar 2006 17:57:42 -0800 } 4, 19 -- Received: from localhost (wpchan-at-localhost) } 4, 19 -- by homer23.u.washington.edu } (8.13.5+UW05.10/8.13.5+Submit) with ESMTP id } k221vf6I025767 } 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 } Mar 2006 17:57:41 -0800 } 4, 19 -- Date: Wed, 1 Mar 2006 17:57:41 -0800 (PST) } 4, 19 -- From: "W. Chan" {wpchan-at-u.washington.edu} } 4, 19 -- To: microscopy-at-microscopy.com } 4, 19 -- Subject: viral particles } 4, 19 -- Message-ID: } {Pine.LNX.4.64.0603011722450.24739-at-homer23.u.washington.edu} } 4, 19 -- MIME-Version: 1.0 } 4, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; } format=flowed } 4, 19 -- X-Uwash-Spam: Gauge=IIIIIII, } Probability=7%, Report='__C230066_P5 0, __CT 0, } __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY } 0, __MIME_VERSION 0, __SANE_MSGID 0' } ==============================End of - } Headers============================== }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 11, 19 -- From drteddunne-at-yahoo.com Thu Mar 2 09:23:29 2006 11, 19 -- Received: from web33411.mail.mud.yahoo.com (web33411.mail.mud.yahoo.com [68.142.206.143]) 11, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k22FNSpZ012360 11, 19 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 09:23:29 -0600 11, 19 -- Received: (qmail 94792 invoked by uid 60001); 2 Mar 2006 15:23:28 -0000 11, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 19 -- s=s1024; d=yahoo.com; 11, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 11, 19 -- b=U2u1+/qPDDqTnvnvab7y+jwzrwPz1JMTPbDwXwNgBXqT2RpTGiGaNP3FizX7k/xCqlRdoplngnQEZcqPJYZ1hX+VtYUjeWNMrn1ADVQHfZv3MjLjocLVotuGR3ZWFnJqDYS2cIzSqtL/XkuEyRStnB5xeIp1OCl9LsNdWscuvlk= ; 11, 19 -- Message-ID: {20060302152328.94790.qmail-at-web33411.mail.mud.yahoo.com} 11, 19 -- Received: from [202.47.247.156] by web33411.mail.mud.yahoo.com via HTTP; Thu, 02 Mar 2006 07:23:28 PST 11, 19 -- Date: Thu, 2 Mar 2006 07:23:28 -0800 (PST) 11, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 11, 19 -- Subject: Re: [Microscopy] viral particles 11, 19 -- To: microscopy-at-microscopy.com 11, 19 -- In-Reply-To: {200603020311.k223Bp32031503-at-ns.microscopy.com} 11, 19 -- MIME-Version: 1.0 11, 19 -- Content-Type: text/plain; charset=iso-8859-1 11, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I have just gotten this message, but the message I sent had no attachments. Is there any way you can check this? Maybe I have a virus...
Thank you,
dj
On Thu, 2 Mar 2006 spamMfilter-at-microscopy.com wrote:
} Subject: REJECTED MAIL } X-Mailer: MicroscopyListSpam Mail Filter vNJZ-2005080908 } X-lewp: MicroscopyListSpam NAGS } } -------------------------------------------------------- } Your mail has been rejected for the following reason(s): } -------------------------------------------------------- } Other: } } Content-Type: multipart/alternative } } An Email Attachment was detected with your message!!! } The Microscopy Listserver will not accept any ATTACHMENTS as a safety measure. } Suspect or Possibly an Unregistered User Address: dljones-at-bestweb.net } } Site match: verizon.net } --------------------------------------------------------------------- } If you have a legitimate reason to contact this SITE, you may get your } mail through the filter by FORWARDing this Email together with } all the error messages and header linesto the Address: } } } } MicroscopyListSpamFilter-at-Microscopy.Com: } } --------------------------------------------------------------------- } } } MicroscopyList Email Filter vNJZ-2005080908 - Your Friendly Neighborhood SysOp } } The text of the rejected email follows: } ********************************************************************* } } This message is in MIME format. The first part should be readable text, } while the remaining parts are likely unreadable without MIME-aware tools. } } --6400082-538-1141314083=:1576 } Content-Type: TEXT/PLAIN; charset=X-UNKNOWN; format=flowed } Content-Transfer-Encoding: QUOTED-PRINTABLE } } Giovanni, } } I do have a video for an Amray 1100 SEM. I haven't looked at it in a long t= } ime=20 } but I believe it has a lot of fundamentals of electron microscopy and EDS= } =20 } operation that is independent of instrument, although the AMR 1100 is quite= } =20 } similar to the AMR 1000 you are asking for. In fact, if you could send me a= } =20 } photo of the main control panel, I could probably indicate the differences = } and=20 } similarities in the two. } } I don't recall if it has much about sample preperation, but if it did, it w= } ould=20 } likely be aimed at metallurgy and not biological samples, which are probabl= } y=20 } what you would prefer. But I'm sure you can more easily find sample prep in= } fo=20 } focused towards biology fairly easily. } } The video I have is neither in VHS or DVD, it is in Video 8. I will have to= } =20 } figure out how to convert these tapes. } } I would be willing to copy them and send them to you for the base cost of= } =20 } materials and shipping. } } Let me know how you would like to proceed. } } dj } } On Wed, 1 Mar 2006 exploratorium-at-tiscali.it wrote: } } --| Email: exploratorium-at-tiscali.it } --| Name: giovanni de caro } --| } --| Organization: associazione luigi montalb=DA } --| } --| Education: 9-12th Grade High School } --| } --| Location: Campobasso, Italy } --| } --| Title: SEM operation video } --| } --| Question: I am looking for a video (VHS or DVD) dealing with the practica= } l=20 } --| operation of an older SEM. We are going to restart and operate an AMR 100= } 0 SEM=20 } --| in our natural science museum for youngsters in Italy=20 } --| (http://web.tiscali.it/exploratorium). Of course I do not expect a docume= } ntary=20 } --| ilustrating THIS specific instrument, I would be more than happy of somet= } hing=20 } --| showing the operation of this calss of instruments. I also would like to = } have=20 } --| a video on SEM specimen preparation. Thank you for your kind help. } --6400082-538-1141314083=:1576-- } } } ==============================Original Headers============================== } 12, 25 -- From dljones-at-bestweb.net Thu Mar 2 09:31:56 2006 } 12, 25 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) } 12, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22FVtKf014572 } 12, 25 -- for |--Microscopy-at-microscopy.com--|; Thu, 2 Mar 2006 09:31:56 -0600 } 12, 25 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) } 12, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id C1DB41CCF0; } 12, 25 -- Thu, 2 Mar 2006 10:31:54 -0500 (EST) } 12, 25 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) } 12, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id 400E61CCCB; } 12, 25 -- Thu, 2 Mar 2006 10:31:54 -0500 (EST) } 12, 25 -- Date: Thu, 2 Mar 2006 10:41:23 -0500 (Eastern Standard Time) } 12, 25 -- From: "David L. Jones" |--dljones-at-bestweb.net--| } 12, 25 -- To: exploratorium-at-tiscali.it } 12, 25 -- cc: Microscopy-at-microscopy.com } 12, 25 -- Subject: Re: [Microscopy] AskAMicroscopist: SEM operation video } 12, 25 -- In-Reply-To: |--200603020141.k221fcTU001827-at-ns.microscopy.com--| } 12, 25 -- Message-ID: |--Pine.WNT.4.62.0603021029330.1576-at-dlj--| } 12, 25 -- References: |--200603020141.k221fcTU001827-at-ns.microscopy.com--| } 12, 25 -- X-X-Sender: dljones-at-pop-croton.bestweb.net } 12, 25 -- MIME-Version: 1.0 } 12, 25 -- Content-Type: MULTIPART/MIXED; BOUNDARY="6400082-538-1141314083=:1576" } 12, 25 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net } 12, 25 -- X-Spam-Level: } 12, 25 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed } 12, 25 -- version=3.0.2 } ==============================End of - Headers============================== }
==============================Original Headers============================== 7, 24 -- From dljones-at-bestweb.net Thu Mar 2 09:39:53 2006 7, 24 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) 7, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22FdqZQ016721 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 09:39:52 -0600 7, 24 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) 7, 24 -- by smtp2.bestweb.net (Postfix) with ESMTP id DC68A1CCC3 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 10:39:51 -0500 (EST) 7, 24 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) 7, 24 -- by smtp2.bestweb.net (Postfix) with ESMTP id 570F21CC37 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 10:39:51 -0500 (EST) 7, 24 -- Date: Thu, 2 Mar 2006 10:49:20 -0500 (Eastern Standard Time) 7, 24 -- From: "David L. Jones" {dljones-at-bestweb.net} 7, 24 -- To: Microscopy-at-microscopy.com 7, 24 -- Subject: Re: AskAMicroscopist: SEM operation video 7, 24 -- In-Reply-To: {200603021531.k22FVwuC014586-at-ns.microscopy.com} 7, 24 -- Message-ID: {Pine.WNT.4.62.0603021048000.1576-at-dlj} 7, 24 -- References: {200603021531.k22FVwuC014586-at-ns.microscopy.com} 7, 24 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 7, 24 -- MIME-Version: 1.0 7, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 7, 24 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net 7, 24 -- X-Spam-Level: 7, 24 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed 7, 24 -- version=3.0.2 ==============================End of - Headers==============================
If the student is looking for macromolecules you actually need an angle of 8 to10 degrees for Pt and 90 degrees for Carbon. If he uses high angles like 45 he will not see anything. Those high angles were used for big stuff like Bacteria and some of the larger viruses like TMV. I have several protocols that I can supply off line.
Best, Al Coritz, Technical Engineer Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- X-from: {gwe-at-ufl.edu} To: {Sampleprep-at-earthlink.net} Sent: Wednesday, March 01, 2006 3:04 PM
Colleagues, As I have not seen a post on the subject of Rob Apkarian's accidental death yesterday I thought I would convey the sad news to the list. I did not know Rob well but we met and interacted at M&M meetings over many years. Rob had enthusiasm in abundance for electron microscopy and I will remember him as a "real character".
Regards
Chris
Christopher J Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.A04 UT Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-9039 Phone +1 214 648 2827 Fax +1 214 648 6408
==============================Original Headers============================== 5, 21 -- From christopher.gilpin-at-utsouthwestern.edu Thu Mar 2 15:16:25 2006 5, 21 -- Received: from swlx167.swmed.edu (swlx167.swmed.edu [199.165.152.167]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22LGO6o023123 5, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 15:16:24 -0600 5, 21 -- Message-Id: {200603022116.k22LGO6o023123-at-ns.microscopy.com} 5, 21 -- Received: from [129.112.148.58] (helo=cgdesktop) 5, 21 -- by swlx167.swmed.edu with esmtp (Exim 4.44) 5, 21 -- id 1FEv9s-0001YB-3p 5, 21 -- for Microscopy-at-microscopy.com; Thu, 02 Mar 2006 15:16:24 -0600 5, 21 -- From: "Christopher Gilpin" {christopher.gilpin-at-utsouthwestern.edu} 5, 21 -- To: {Microscopy-at-microscopy.com} 5, 21 -- Date: Thu, 2 Mar 2006 15:16:36 -0600 5, 21 -- MIME-Version: 1.0 5, 21 -- Content-Type: text/plain; 5, 21 -- charset="us-ascii" 5, 21 -- Content-Transfer-Encoding: 7bit 5, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 5, 21 -- Thread-Index: AcY+PpbOIka7Tyf2TCO7Qu1UmfNBBA== 5, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 5, 21 -- X-Scan-Signature: ab3f4c55490e4d672805895add812bc5 5, 21 -- Subject: Rob Apakrian passed away ==============================End of - Headers==============================
Thanks for the feedback on dealing with low eV peak pileup.
It looks like Ni and Pd are good options. Does anyone have experience with these and know if they oxidize?
Ni is 99.9% pure and is .062" thick. Pd is 99.5% pure and can be .004" or .008" thick. So I guess that Ni deposits faster than Pd. I recall the typical thickness for Au/Pd is about .01".
I don't think that just one target type will work for all specimens. That is OK. Changing targets is not all that big of a deal.
Thanks for the help.
gary g.
==============================Original Headers============================== 7, 17 -- From gary-at-gaugler.com Thu Mar 2 16:06:15 2006 7, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k22M6E0b000319 7, 17 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 16:06:14 -0600 7, 17 -- Received: (qmail 23885 invoked from network); 2 Mar 2006 14:06:05 -0800 7, 17 -- Received: by simscan 1.1.0 ppid: 23882, pid: 23883, t: 0.1081s 7, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 7, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 7, 17 -- by qsmtp2 with SMTP; 2 Mar 2006 14:06:04 -0800 7, 17 -- Message-Id: {6.2.3.4.2.20060302135842.01ff6768-at-mail.calweb.com} 7, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 7, 17 -- Date: Thu, 02 Mar 2006 14:06:14 -0800 7, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 7, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 7, 17 -- Subject: Ni and Pd target material 7, 17 -- Mime-Version: 1.0 7, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
With deep sorrow, I am passing onto the list the tragic news of Dr. Rob Apkarian's Death, which occurred in Atlanta in the evening of February 28, 2006. Many of you may know Rob for his remarkable work in cryo-EM and high resolution SEM. Some of you may also have active collaborations with Rob. Rob was the director of Integrated Microscopy and Microanalytical Facility in Emory and a close colleague of mine in Emory microscopy research community. Rob was also an active member of the MSA leadership for many years.
Currently, I do not know about any plan regarding any religious service or funeral yet. But if anyone wishes to send a condolence card or flowers, I can help you get connected if you contact me off-line. Rob is survived by his wife who is also a member of Emory community.
Take care and live well.
Hong Emory EM
==============================Original Headers============================== 10, 18 -- From hyi-at-emory.edu Thu Mar 2 17:24:13 2006 10, 18 -- Received: from rwcrmhc11.comcast.net (rwcrmhc11.comcast.net [216.148.227.151]) 10, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22NOCjx011488 10, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 17:24:12 -0600 10, 18 -- Received: from [24.30.86.130] (c-24-30-86-130.hsd1.ga.comcast.net[24.30.86.130]) 10, 18 -- by comcast.net (rwcrmhc11) with SMTP 10, 18 -- id {20060302232404m11009rnpie} ; Thu, 2 Mar 2006 23:24:08 +0000 10, 18 -- Mime-Version: 1.0 (Apple Message framework v622) 10, 18 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 18 -- Message-Id: {8cdae9f5fb505007537fe576bc1aeeb9-at-emory.edu} 10, 18 -- Cc: BusinessOffice-at-microscopy.org 10, 18 -- From: Hong Yi {hyi-at-emory.edu} 10, 18 -- Subject: (Microscopy) Dr. Apkarian 10, 18 -- Date: Thu, 2 Mar 2006 18:24:10 -0500 10, 18 -- To: Microscopy-at-microscopy.com 10, 18 -- X-Mailer: Apple Mail (2.622) 10, 18 -- Content-Transfer-Encoding: 8bit 10, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k22NOCjx011488 ==============================End of - Headers==============================
Thank you Chris and Hong for the posts about Rob's passing. A service for Rob is being planned for next week, the details have not been finalized. For those of you wishing to extend your sympathies to Rob's wife, you may contact me off-line as well.
He was a great research scientist, mentor, and a dear friend. I will miss him deeply.
Sincerely,
Elizabeth R. Wright
==============================Original Headers============================== 6, 20 -- From erwright-at-caltech.edu Thu Mar 2 18:16:08 2006 6, 20 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k230G6rA021415 6, 20 -- for {microscopy-at-Microscopy.Com} ; Thu, 2 Mar 2006 18:16:07 -0600 6, 20 -- Received: from localhost (water-dog [192.168.1.26]) 6, 20 -- by fire-ox-postvirus (Postfix) with ESMTP 6, 20 -- id 1598435E88; Thu, 2 Mar 2006 16:16:06 -0800 (PST) 6, 20 -- Received: from [192.168.157.114] (pix-1.caltech.edu [131.215.2.21]) 6, 20 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP 6, 20 -- id 112C535C32; Thu, 2 Mar 2006 16:16:05 -0800 (PST) 6, 20 -- Mime-Version: 1.0 (Apple Message framework v623) 6, 20 -- Content-Transfer-Encoding: 7bit 6, 20 -- Message-Id: {ee1d05f1abe4f266fe60b5bc3cb54fea-at-caltech.edu} 6, 20 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 6, 20 -- To: microscopy-at-Microscopy.Com, 3dem-at-ucsd.edu 6, 20 -- From: "Elizabeth R.Wright" {erwright-at-caltech.edu} 6, 20 -- Subject: Dr. Robert P. Apkarian 6, 20 -- Date: Thu, 2 Mar 2006 16:16:04 -0800 6, 20 -- X-Mailer: Apple Mail (2.623) 6, 20 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mariac-h-at-uniandes.edu.co) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 2, 2006 at 18:29:31 ---------------------------------------------------------------------------
Email: mariac-h-at-uniandes.edu.co Name: Maria Cristina Herrera
Organization: Universidad de Los Andes
Education: Graduate College
Location: Bogota, Colombia
Question: I got some SEM images of soils. I can identify some minerals but I am not sure about somethings that look like dehidrated roots. The scale of my images is 10 microns. The roots are longer than 10 microns and about 1 micron thick. I would like to know if roots can be this large and if there is somebody who can help me to define what are those things in my images..I can email my images. Thanks.
If the specimen is very small, and/or has low contrast (unstained), and can be deposited onto a formvar or carbon-coated grid, then the student could shadow the sample on-grid with Pt/C and omit the carbon altogether. The carbon layer is only important so that the replica doesn't distort or crack when it is floated off of the substrate. Because there is no extra carbon layer, there is a slight increase in contrast with an on-grid shadow vs a true replica. As mentioned in previous replies, small samples usually look best when shadowed at a low angle (10-15 deg. from horizontal). Furthermore, rotary shadowing usually looks best on fibers while fixed-angle shadowing looks best on globular/particulate specimens. And finally, there are less steps involved in making an on-grid shadow vs a replica.
Sorry if we've beat this topic to death,
Andrew Bowling The University of Texas at Austin
==============================Original Headers============================== 6, 17 -- From abowling-at-mail.utexas.edu Thu Mar 2 18:48:04 2006 6, 17 -- Received: from wb6-a.mail.utexas.edu (wb6-a.mail.utexas.edu [128.83.126.144]) 6, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k230m2nb028499 6, 17 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 18:48:02 -0600 6, 17 -- Received: (qmail 22244 invoked from network); 3 Mar 2006 00:48:01 -0000 6, 17 -- Received: from dhcp-146-6-151-204.biosci.utexas.edu (HELO ?146.6.151.204?) (abowling-at-146.6.151.204) 6, 17 -- by wb6.mail.utexas.edu with (RC4-MD5 encrypted) ESMTPSA; 3 Mar 2006 00:48:01 -0000 6, 17 -- Message-ID: {440791F4.2080606-at-mail.utexas.edu} 6, 17 -- Date: Thu, 02 Mar 2006 18:46:44 -0600 6, 17 -- From: Andrew Bowling {abowling-at-mail.utexas.edu} 6, 17 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 6, 17 -- X-Accept-Language: en-us, en 6, 17 -- MIME-Version: 1.0 6, 17 -- To: Microscopy-at-microscopy.com 6, 17 -- Subject: [Microscopy] Re: Replicas 6, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (andromeda_tm-at-libero.it) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 3, 2006 at 04:32:02 ---------------------------------------------------------------------------
Hi all, does someone know the protocol for staining with the ìEhrlich Biondi Heidenhainî method? Particularly how long has the specimen to stay in the staining solution? I have noticed, in previous, tests that is quit difficult to dye the nucleus with the methyl green contained in the solution. I am processing thin sections of mouse liver included in wax and fixed with Bouin liquid. I also wonder why the sections are crunched after cutting. There is a solution at this problem? Thanks in advance. Best Regards, Massimo
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (andromeda_tm-at-libero.it) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 3, 2006 at 04:36:47 ---------------------------------------------------------------------------
Hi everyone, would any of you give me some information about the procedure on using Aceto Carmine for nucleos staining and what can I use like contrast colour? Thank you in advance for your assistance. Best Regards, Massimo
I would like to contact anyone who has experience working with tobacco suspension cells. I am interested in general morphology information as well as preparation using standard chemical fix and high pressure freezing methods.
As this is a rather specific request, you might want to contact me off-line.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 5, 21 -- From dsherman-at-purdue.edu Fri Mar 3 08:46:44 2006 5, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k23EkhFF017338 5, 21 -- for {microscopy-at-microscopy.com} ; Fri, 3 Mar 2006 08:46:43 -0600 5, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.235]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 5, 21 -- Fri, 3 Mar 2006 09:46:41 -0500 5, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 5, 21 -- Fri, 3 Mar 2006 14:46:42 +0000 5, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 5, 21 -- Date: Fri, 03 Mar 2006 09:46:41 -0500 5, 21 -- Subject: Tobacco suspension cells 5, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 5, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 5, 21 -- Message-ID: {C02DC101.DCF0%dsherman-at-purdue.edu} 5, 21 -- Thread-Topic: Tobacco suspension cells 5, 21 -- Thread-Index: AcY+0UhghzF7x6rEEdq0gQARJN08Mg== 5, 21 -- Mime-version: 1.0 5, 21 -- Content-type: text/plain; 5, 21 -- charset="US-ASCII" 5, 21 -- Content-transfer-encoding: 7bit 5, 21 -- X-OriginalArrivalTime: 03 Mar 2006 14:46:41.0564 (UTC) FILETIME=[48B66DC0:01C63ED1] ==============================End of - Headers==============================
The following is the information regarding the funeral service for Dr. Apkarian. You can also view the announcement at http://www.chemistry.emory.edu/. Thank you.
Hong Emory EM
} We regret to announce that Dr. Robert Apkarian died in an traffic } accident on Tuesday, February 28, 2006. The Department of Chemistry } extends our deepest condolences to the Apkarian family. } } The funeral will be Monday, March 6th. } at the } Greek Orthodox Church } 2480 Clairmont Road, NE } Atlanta, GA 30329 } } 10:00 to 11:00 a.m. Viewing } 11:00 - 12:00 Service } Reception immediately following } } } All are welcome to attend. } } Please assist us by forwarding this information to any of Dr. } Apkarian's } friends who we may not have reached. } } Thank you.
==============================Original Headers============================== 7, 21 -- From hyi-at-emory.edu Fri Mar 3 09:01:29 2006 7, 21 -- Received: from pales.cc.emory.edu (pales.cc.emory.edu [170.140.8.221]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k23F1SWm021950 7, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Mar 2006 09:01:28 -0600 7, 21 -- Received: from [170.140.233.152] (localhost [127.0.0.1]) 7, 21 -- by pales.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k23F1RlK014558 7, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Mar 2006 10:01:27 -0500 (EST) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v622) 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- Message-Id: {96f4f9509ca6223516484dc8bdf4b16d-at-emory.edu} 7, 21 -- Content-Type: text/plain; 7, 21 -- charset=US-ASCII; 7, 21 -- format=flowed 7, 21 -- To: Microscopy-at-microscopy.com 7, 21 -- From: Hong Yi {hyi-at-emory.edu} 7, 21 -- Subject: Fwd: [Fwd: Dr. Rob Apkarian] 7, 21 -- Date: Fri, 3 Mar 2006 10:01:26 -0500 7, 21 -- X-Mailer: Apple Mail (2.622) 7, 21 -- X-imss-version: 2.038 7, 21 -- X-imss-result: Passed 7, 21 -- X-imss-approveListMatch: *-at-emory.edu ==============================End of - Headers==============================
We are having trouble with our old LifeCell freezing machine (a slam freezer) so we would be interested in obtaining one that someone has laying around in the corner. It does not even have to be functioning as we have some clever folks here who might be able to fix ours with old parts. Please contact me with any ideas, machines or parts that might be out there somewhere.
There is a job open at Children's Hospital San Diego for a TEM/Histologist. The person will be responsible for doing the majority of EM (the EM workload varies, though it is almost exclusively renal tissue) and assisting in the Histology section when not doing EM (which happens frequently). So histology skills are an obvious plus.
If you are interested please contact me at psicurello-at-chsd.org or Dr. Eric Breisch at ebreisch-at-chsg.org. Please include a copy of your CV or resume.
Sunny San Diego awaits you!
Paula Sicurello :-)
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 3, 18 -- From patpxs-at-yahoo.com Fri Mar 3 12:09:00 2006 3, 18 -- Received: from web52202.mail.yahoo.com (web52202.mail.yahoo.com [206.190.48.125]) 3, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k23I8xTd000725 3, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Mar 2006 12:08:59 -0600 3, 18 -- Received: (qmail 2020 invoked by uid 60001); 3 Mar 2006 18:08:59 -0000 3, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 3, 18 -- s=s1024; d=yahoo.com; 3, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 3, 18 -- b=K/KClLe8EWmfVPKcUH9yRpeaPR7Tb2H5gmmoQQ66aLtQrbP98K5C2hRzZ9mW0ERYeDHLoh3/pxARyc4IUUb5kvJmhCmdKTW/0cT5GyjqGJKJPuDEMLi6aEmtMhiiDLnLokMFx7DAqMEzrTOifo1OJ+BaIO6zhfBivbFMN8vx7UE= ; 3, 18 -- Message-ID: {20060303180859.2018.qmail-at-web52202.mail.yahoo.com} 3, 18 -- Received: from [209.203.68.199] by web52202.mail.yahoo.com via HTTP; Fri, 03 Mar 2006 10:08:59 PST 3, 18 -- Date: Fri, 3 Mar 2006 10:08:59 -0800 (PST) 3, 18 -- From: Paula Sicurello {patpxs-at-yahoo.com} 3, 18 -- Subject: TEM/Histologist position open in San Diego 3, 18 -- To: MSA BB {microscopy-at-msa.microscopy.com} 3, 18 -- MIME-Version: 1.0 3, 18 -- Content-Type: text/plain; charset=iso-8859-1 3, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both fremingt-at-fhcrc.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: We are asking if anyone has a V. Avarlaid Autoradiographic slide coating machine they would like to unload? Parts of ours are missing and they are no longer made. Replys can be made to me at fremingt-at-fhcrc.org. Thanks.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both gibi55-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: gibi55-at-yahoo.com Name: Gino Bianchi
Organization: Universidad Central de Venezuela
Title-Subject: [Filtered] Autotechnicon wanted
Question: Looking for an Autotechnicon mono or duo in working order
I'm with Balzers, Inc., a supplier of hard coating equipment and services. We have just opened an Applications Support Center in Elgin, IL. We are in need (hopefully temporarily) of SEM service nearby.
If anyone knows of or supplies service in this area, I would appreciate this in formation.
Best regards,
Erik
C. Erik Bauer Tool Coating Specialist Balzers, Inc. Applications Support Center 1181 Jansen Farms Ct. Elgin, IL 60123 tel: 847-695-5200 ext.2001 cell: 224-730-084 fax: 847-695-4051 erik.bauer-at-balzers.com www.balzers.com
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 8, 20 -- From cerikbauer-at-yahoo.com Mon Mar 6 08:31:50 2006 8, 20 -- Received: from web60421.mail.yahoo.com (web60421.mail.yahoo.com [209.73.178.149]) 8, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k26EVmCG015478 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 6 Mar 2006 08:31:48 -0600 8, 20 -- Received: (qmail 11744 invoked by uid 60001); 6 Mar 2006 14:31:48 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=Nim4mrtWgLFwHgT2ibLpYVAfN0b75xTX+I0fBLsBpxSVAD6+8Q0SOW/4vwLJh/5jJLElN/O8yYV4O2cfDQaVUa4fuDMFY6IpH9oP+Zpjv8q5jbsMhXnx5XWQFFhcqPOsI6S+0sB+nL7OxkvVxcVSOjrJEyVCY5rJbLEE9aonQlI= ; 8, 20 -- Message-ID: {20060306143148.11742.qmail-at-web60421.mail.yahoo.com} 8, 20 -- Received: from [63.144.89.98] by web60421.mail.yahoo.com via HTTP; Mon, 06 Mar 2006 06:31:48 PST 8, 20 -- Date: Mon, 6 Mar 2006 06:31:48 -0800 (PST) 8, 20 -- From: Erik Bauer {cerikbauer-at-yahoo.com} 8, 20 -- Subject: SEM Services near Chicago 8, 20 -- To: List Microscopy {Microscopy-at-MSA.Microscopy.Com} 8, 20 -- Cc: "Dennis T. Quinto" {dennis.quinto-at-balzers.com} , 8, 20 -- volker.derflinger-at-balzers.com 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
We will have a full time job opening for an entry level EM Tech. This position will support our current operation for clinical (hospital) as well as our research needs (medical school).
Thank you.
Rajesh Patel Imaging Suite Rm 024 School of Public Health 683 Hoes Lane Piscataway, NJ 08854
Just a word of caution, there are health/safety issues in dealing with inorganic fluorine compounds. Make sure you find the proper MSDS's for what you're dealing with.
Jane L. LaGoy Lab Manager/R&D Engineer Bodycote North America 155 River Street Andover, MA 01810 978-470-1620 x450 FAX: 978-475-2951 jane.lagoy-at-bodycote.com The only people to get even with are those who have helped you.
-----Original Message----- X-from: randy-nessler-at-uiowa.edu [mailto:randy-nessler-at-uiowa.edu] Sent: Tuesday, February 28, 2006 6:06 PM To: jane.lagoy-at-bodycote.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying
please copy both randy-nessler-at-uiowa.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: randy-nessler-at-uiowa.edu Name: Randy Nessler
Organization: University of Iowa
Title-Subject: [Filtered] Fluorine removal?
Question: I have been asked to post this question for a collegue. It appears that he might be getting fluorine contamination of his XPS samples when they are in a processing chamber. The chamber is lined with glass. Is there a suitable reagent to clean this chamber out with to remove any residual fluorine? "Baking" the chamber hasn't minimied the problem. Thanks, Randy
The seventh annual European ESEM Userclub meeting will take place in London on June 26th 2006, preceeding the Royal Microscopical Society's MICROSCIENCE 2006 conference.
For more information & registration, please visit:
http://www.rms.org.co.uk/events_esem/shtml
(By the way, to register for MICROSCIENCE 2006, June 27th - 29th, please visit http://www.microscience2006.org.uk/conference_registration.shtml. Note that the abstract deadline is 3rd April 2006).
Best Wishes,
Debbie.
-- Dr Debbie Stokes
Biological & Soft Systems University of Cambridge Dept of Physics Cavendish Laboratory JJ Thomson Avenue Cambridge CB3 0HE
I need to glue a little plastic ring to the end of a TEM cryoholder. Armstrong A12 was recommended, but I don't have any around. Anyone know where I can get some or if any epoxy is OK? Especially concerned about something that is good with vacuum and cryo temps.
I have some old M-bond 610 around that might do, but the directions say its shelf life is only about 9 months even unmixed. Mine is older than that, is it really no good?
Thanks
Jon -- Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 5, 21 -- From jmkrupp-at-ucsc.edu Mon Mar 6 16:44:18 2006 5, 21 -- Received: from smtp-prod-mx2.ucsc.edu (smtp-prod-mx2.ucsc.edu [128.114.125.44]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k26MiHeF006991 5, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Mar 2006 16:44:17 -0600 5, 21 -- Received: from ucsc.edu (cruzmail-fe1.ucsc.edu [128.114.125.5]) 5, 21 -- by smtp-prod-mx2.ucsc.edu (8.13.1/8.13.1) with ESMTP id k26MaAfw024370 5, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Mar 2006 14:36:11 -0800 (PST) 5, 21 -- Received: from [128.114.25.190] (account jmkrupp-at-ucsc.edu [128.114.25.190] verified) 5, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 5, 21 -- with ESMTPA id 48085986 for Microscopy-at-Microscopy.Com; Mon, 06 Mar 2006 14:36:05 -0800 5, 21 -- Mime-Version: 1.0 5, 21 -- Message-Id: {p06230903c032684ff6f1-at-[128.114.25.190]} 5, 21 -- Date: Mon, 6 Mar 2006 14:36:04 -0800 5, 21 -- To: Microscopy-at-microscopy.com 5, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 5, 21 -- Subject: Armstrong A12 5, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 5, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 5, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 5, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
As a brand new Hitachi user (S-3400N II), is was somewhat amazed to find that there is no dedicated Hitachi user listserver.
I also run a Cameca SX51 electron probe and with several other users in 1994 initiated the sx50-users listserver that has been infinitely useful for the past 12 years. And a JEOL epma list started a couple of years ago.
So why no listserver for Hitachi SEM users? It would seem that it would be quite useful for both new and experienced users, and helpful for sorting out a variety of issues and problems specific to users of these flavor instruments.
If you are interested, please contact me off line (off the list), direct to johnf-at-geology.wisc.edu
John Fournelle -- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
==============================Original Headers============================== 7, 24 -- From johnf-at-geology.wisc.edu Mon Mar 6 18:02:33 2006 7, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 7, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2702XOU026881 7, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 6 Mar 2006 18:02:33 -0600 7, 24 -- Received: from localhost (localhost [127.0.0.1]) 7, 24 -- by localhost (Postfix) with ESMTP id 0663620D1A 7, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 6 Mar 2006 18:02:33 -0600 (CST) 7, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 7, 24 -- by localhost (ice [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 7, 24 -- id 05536-09 for {Microscopy-at-Microscopy.Com} ; 7, 24 -- Mon, 6 Mar 2006 18:02:30 -0600 (CST) 7, 24 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 7, 24 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 7, 24 -- (No client certificate requested) 7, 24 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 7C5C320D0C 7, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 6 Mar 2006 18:02:30 -0600 (CST) 7, 24 -- Mime-Version: 1.0 7, 24 -- Message-Id: {p06230906c0327be78b16-at-[144.92.206.57]} 7, 24 -- Date: Mon, 6 Mar 2006 17:59:44 -0600 7, 24 -- To: Microscopy List_nestors {Microscopy-at-Microscopy.Com} 7, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 7, 24 -- Subject: Hitachi SEM Users? 7, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 24 -- X-Virus-Scanned: by amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
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Email: Dorothy.Howard-at-noaa.gov Name: Dorothy Howard
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Email: fulton.2-at-osu.edu Name: Dave Fulton
Organization: OARDC/MCIC/Ohio State University
Title-Subject: [Filtered] Microwave tissue processing for TEM
Question: Hello fellow Listers; Does anyone out there have a Microwave Research and Applications, Inc. , model BP-111-RS laboratory microwave oven, which they are using to process tissue samples for TEM? We have recently purchased one and have made several attempts to use it to process plant tissue (maize and tobacco) for TEM without success. If you have successful protocols that you have used with this oven and would be willing to share them, it would be greatly appreciated. You may reply directly to me if you wish. Thanks in advance for your time and trouble.
Dorothy Howard wrote: ========================================== Question: Could you tell me where in Maryland or Delaware you can contract for EM services? ==========================================
Would "just over the border in Pennsylvania" be considered? Structure Probe, Inc. has been offering both SEM and TEM services since 1970 as a laboratory service in West Chester, PA. Our contact information if given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 14, 25 -- From cgarber-at-2spi.com Mon Mar 6 22:32:44 2006 14, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 14, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k274WgUX017564 14, 25 -- for {microscopy-at-msa.microscopy.com} ; Mon, 6 Mar 2006 22:32:43 -0600 14, 25 -- Received: from ibm1x23g2abfyg (217-118-122-167.client.stsn.net [217.118.122.167]) 14, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k274WdIY015703; 14, 25 -- Mon, 6 Mar 2006 23:32:41 -0500 14, 25 -- X-IDV-FirstRcvd: 217-118-122-167.client.stsn.net [217.118.122.167] 14, 25 -- X-IDV-HELO: ibm1x23g2abfyg 14, 25 -- Message-ID: {02b101c641a0$298ac190$4b3f680a-at-ibm1x23g2abfyg} 14, 25 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 14, 25 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 14, 25 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 14, 25 -- Cc: {Dorothy.Howard-at-noaa.gov} 14, 25 -- Subject: TEM and SEM contract services in DE and MD 14, 25 -- Date: Mon, 6 Mar 2006 23:32:35 -0500 14, 25 -- MIME-Version: 1.0 14, 25 -- Content-Type: text/plain; 14, 25 -- charset="Windows-1252" 14, 25 -- X-Priority: 3 14, 25 -- X-MSMail-Priority: Normal 14, 25 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 14, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 14, 25 -- Content-Transfer-Encoding: 8bit 14, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k274WgUX017564 ==============================End of - Headers==============================
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Email: g.posthuma-at-lab.azu.nl Name: George Posthuma
Organization: Dept of Cell Biology, Utrecht, The netherlands
Title-Subject: [Filtered] workshop on Cryomethods, Ultracryotomy and Immunolabeling
Question: dear all,
In collaboration with Leica we will organize a workshop on Cryomethods, Ultracryotomy and Immunolabeling in Utrecht, The Netherlands. If you are interested, please have a look at: http://www.cmc-utrecht.nl/education/indexeducation.htm. The maximum of participants is 16, of which already 13 are booked.
This is for all you cryo experts out there. I was contacted by a researcher who wants to look at ice crystals in thin sections of ice cream in the TEM. Anyone out there who has the capability to do this sort of cryo work? The researcher is located at the University of Nebraska-Lincoln, Lincoln, NE. We do not have the cryo capability here in Nebraska. So I would like to find out who out there is closest and then we'll see if we can find a way to get the ice cream to you without it melting. This individual is not an EM person so I offerred to try to gather some information for him to see what might be available in the way of help. If anyone out there can help, please contact me and I'll pass the information along. Thanks
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center Omaha, NE 68198-6395 e-mail: tbargar-at-unmc.edu phone: 402-559-7347 fax: 402-559-3400
==============================Original Headers============================== 5, 20 -- From tbargar-at-unmc.edu Tue Mar 7 09:19:18 2006 5, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27FJIJs015410 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:19:18 -0600 5, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 5, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id A2F00A00AF 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:15 -0600 (CST) 5, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 5, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 30DAE398085 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:14 -0600 (CST) 5, 20 -- Subject: Need cryoultramicrotomy of ice cream 5, 20 -- To: Microscopy-at-MSA.Microscopy.com 5, 20 -- X-Mailer: Lotus Notes Release 6.5.1 January 21, 2004 5, 20 -- Message-ID: {OFA994B2B5.45E530B5-ON8625712A.0052F932-8625712A.005428D0-at-unmc.edu} 5, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 5, 20 -- Date: Tue, 7 Mar 2006 09:19:14 -0600 5, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR(Release 6.5.5 5, 20 -- HF62|January 19, 2006) at 03/07/2006 09:19:16 AM 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Um ... any particular reason for the thin sections? CryoSEM of ice cream has been done to examine ice crystals, lipid droplets, and air spaces. I'd think this would be a more useful (and much less stressful) technique than is cryoultrasectioning and cryo TEM and opening the crystals don't change in the process, then watching them change in the beam ... Lincoln Lim did this with the cryo stage on the Hitachi S-900 at UW-Madison, using low kV (~1.5, if I remember right), the researcher might want to look for his publications. Are there any cryoSEM labs around Lincoln?
Phil
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==============================Original Headers============================== 3, 23 -- From oshel1pe-at-cmich.edu Tue Mar 7 10:22:22 2006 3, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27GMMGG025898 3, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 10:22:22 -0600 3, 23 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 3, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k27H0r4l012285; 3, 23 -- Tue, 7 Mar 2006 12:00:53 -0500 3, 23 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 3, 23 -- Tue, 7 Mar 2006 11:20:51 -0500 3, 23 -- Mime-Version: 1.0 3, 23 -- Message-Id: {f06230902c03362a1cb3f-at-[141.209.160.132]} 3, 23 -- In-Reply-To: {200603071604.k27G4guS024098-at-ns.microscopy.com} 3, 23 -- References: {200603071604.k27G4guS024098-at-ns.microscopy.com} 3, 23 -- Date: Tue, 7 Mar 2006 11:22:17 -0500 3, 23 -- To: tbargar-at-unmc.edu 3, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 23 -- Subject: Re: [Microscopy] Need cryoultramicrotomy of ice cream 3, 23 -- Cc: Microscopy-at-microscopy.com 3, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 23 -- X-OriginalArrivalTime: 07 Mar 2006 16:20:51.0689 (UTC) FILETIME=[1A1A1590:01C64203] 3, 23 -- X-CanItPRO-Stream: default 3, 23 -- X-Spam-Score: -4 () L_EXCH_MF 3, 23 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Check out the "foods under the microscope" web page: http://www.magma.ca/~scimat/ This is a good resource on a variety of microscopic techniques on dairy products. They discuss the microscopy of yogurt and cheeses, as well as dried milk products. The bibliography is extensive.
Karl -----Original Message----- X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu] Sent: Tuesday, March 07, 2006 10:39 AM To: Karl Hagglund
This is for all you cryo experts out there. I was contacted by a researcher who wants to look at ice crystals in thin sections of ice cream in the TEM. Anyone out there who has the capability to do this sort of cryo work? The researcher is located at the University of Nebraska-Lincoln, Lincoln, NE. We do not have the cryo capability here in Nebraska. So I would like to find out who out there is closest and then we'll see if we can find a way to get the ice cream to you without it melting. This individual is not an EM person so I offerred to try to gather some information for him to see what might be available in the way of help. If anyone out there can help, please contact me and I'll pass the information along. Thanks
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center Omaha, NE 68198-6395 e-mail: tbargar-at-unmc.edu phone: 402-559-7347 fax: 402-559-3400
==============================Original Headers============================== 5, 20 -- From tbargar-at-unmc.edu Tue Mar 7 09:19:18 2006 5, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27FJIJs015410 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:19:18 -0600 5, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 5, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id A2F00A00AF 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:15 -0600 (CST) 5, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 5, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 30DAE398085 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:14 -0600 (CST) 5, 20 -- Subject: Need cryoultramicrotomy of ice cream 5, 20 -- To: Microscopy-at-MSA.Microscopy.com 5, 20 -- X-Mailer: Lotus Notes Release 6.5.1 January 21, 2004 5, 20 -- Message-ID: {OFA994B2B5.45E530B5-ON8625712A.0052F932-8625712A.005428D0-at-unmc.edu} 5, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 5, 20 -- Date: Tue, 7 Mar 2006 09:19:14 -0600 5, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR(Release 6.5.5 5, 20 -- HF62|January 19, 2006) at 03/07/2006 09:19:16 AM 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 13, 24 -- From hagglundk1-at-nku.edu Tue Mar 7 13:46:27 2006 13, 24 -- Received: from mailfe2.nku.edu (exchange.nku.edu [192.122.237.68]) 13, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27JkRpE006150 13, 24 -- for {microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 13:46:27 -0600 13, 24 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 13, 24 -- Tue, 7 Mar 2006 14:46:26 -0500 13, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 13, 24 -- Content-class: urn:content-classes:message 13, 24 -- MIME-Version: 1.0 13, 24 -- Content-Type: text/plain; 13, 24 -- charset="us-ascii" 13, 24 -- Subject: RE: [Microscopy] Need cryoultramicrotomy of ice cream 13, 24 -- Date: Tue, 7 Mar 2006 14:46:26 -0500 13, 24 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F358691-at-mailfac1.hh.nku.edu} 13, 24 -- X-MS-Has-Attach: 13, 24 -- X-MS-TNEF-Correlator: 13, 24 -- Thread-Topic: [Microscopy] Need cryoultramicrotomy of ice cream 13, 24 -- Thread-Index: AcZCALuBxcuDWRkORCi4uvSXIKSRKwAHiweQ 13, 24 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 13, 24 -- To: {tbargar-at-unmc.edu} 13, 24 -- Cc: {microscopy-at-microscopy.com} 13, 24 -- X-OriginalArrivalTime: 07 Mar 2006 19:46:26.0709 (UTC) FILETIME=[D258C450:01C6421F] 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k27JkRpE006150 ==============================End of - Headers==============================
George also comes to the International Cryo-EM course at the University of British Columbia, Vancouver, British Columbia, Canada hosted by Kent McDonald, U Berkeley and Elaine Humphrey, UBC
This is a 10-day course, June 6-15, 2006, covering Cryo-TEM, Cryo-SEM, high pressure freezing, Cryo Ultramicrotomy, Tokuyasu method and immunolabelling.
This year we have new instruments from Leica and Baltec. follow the links from http://www.emlab.ubc.ca
Elaine
} } Email: g.posthuma-at-lab.azu.nl } Name: George Posthuma } } Organization: Dept of Cell Biology, Utrecht, The netherlands } } Title-Subject: [Filtered] workshop on Cryomethods, Ultracryotomy and } Immunolabeling } } Question: dear all, } } In collaboration with Leica we will organize a workshop on } Cryomethods, Ultracryotomy and Immunolabeling in Utrecht, The } Netherlands. If you are interested, please have a look at: } http://www.cmc-utrecht.nl/education/indexeducation.htm. The maximum } of participants is 16, of which already 13 are booked. } } Yours sincerely } } George Posthuma }
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada (2003-2005) University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
==============================Original Headers============================== 7, 30 -- From ech-at-interchange.ubc.ca Tue Mar 7 14:37:18 2006 7, 30 -- Received: from mta3.mail-relay.ubc.ca (mta3.mail-relay.ubc.ca [137.82.45.6]) 7, 30 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27KbGC1015821 7, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Mar 2006 14:37:16 -0600 7, 30 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 7, 30 -- by mta3.mail-relay.ubc.ca (8.12.11/8.12.11) with ESMTP id k27KbDs3025904 7, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Mar 2006 12:37:14 -0800 (PST) 7, 30 -- (envelope-from ech-at-interchange.ubc.ca) 7, 30 -- Received: from [137.82.85.193] (echpowerbook.emlab.ubc.ca [137.82.85.193]) 7, 30 -- by smtp.interchange.ubc.ca 7, 30 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 7, 30 -- with ESMTPA id {0IVR00FP9YLJ63-at-smtp.interchange.ubc.ca} for 7, 30 -- microscopy-at-msa.microscopy.com; Tue, 07 Mar 2006 12:36:55 -0800 (PST) 7, 30 -- Date: Tue, 07 Mar 2006 12:36:54 -0800 7, 30 -- From: Elaine Humphrey {ech-at-interchange.ubc.ca} 7, 30 -- Subject: Re: [Microscopy] viaWWW: workshop on Cryomethods, 7, 30 -- Ultracryotomy and Immunolabeling 7, 30 -- In-reply-to: {200603071425.k27EPhpY007742-at-ns.microscopy.com} 7, 30 -- X-Sender: ech-at-mail.interchange.ubc.ca 7, 30 -- To: microscopy-at-msa.microscopy.com 7, 30 -- Cc: g.posthuma-at-lab.azu.nl 7, 30 -- Message-id: {a06100501c03376ebe414-at-[137.82.85.193]} 7, 30 -- MIME-version: 1.0 7, 30 -- Content-type: text/plain; format=flowed; charset=us-ascii 7, 30 -- References: {200603071425.k27EPhpY007742-at-ns.microscopy.com} 7, 30 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.03.07.115105 7, 30 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 7, 30 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0 7, 30 -- X-Spam-Level: 7, 30 -- X-Spam-Flag: No ==============================End of - Headers==============================
THE TEXAS SOCIETY FOR MICROSCOPY invites you to our Spring 2006 meeting on April 20-22, 2006 at Alcon Research Labs, 6201 South Freeway, Ft. Worth, TX 76134 2ND CALL FOR PAPERS
All registration forms and lodging details are available on our web site: http://www.texasmicroscopy.org/
ABSTRACTS MUST BE RECEIVED BY: March 20, 2006 Advanced Registration Deadline: April 7, 2006. Advanced registration is strongly suggested to afford TSM an accurate participant count for event organization AND to comply with Alcon security requirements. Thursday workshops and Friday sessions are being held at Alcon Research.
**Workshops— Thursday, April 20, 2006
“Microwave Processing: Factors the Influence Results” Sponsored by Ted Pella, Inc. Speaker: Rick Giberson, Sr. Applications Engineer
“ESEM: not just for Biology Anymore” Sponsored by FEI Company Speaker: Daniel Phifer, Sr. Applications Engineer
**Guest Speaker — Friday, April 21, 2006
“Materials Science in Museums” Dr. Pamela Vandiver, Professor of Materials Science and Engineering and Archeology, Co-Director of Program in Heritage Conservation Science at the University of Arizona, and former Senior Research Scientist at the Smithsonian Institution’s Center for Materials Research and Education.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 12, 21 -- From r-holdford-at-ti.com Tue Mar 7 15:43:20 2006 12, 21 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 12, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27LhJZS026508 12, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Mar 2006 15:43:19 -0600 12, 21 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 12, 21 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k27LhAjT009990 12, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Mar 2006 15:43:16 -0600 (CST) 12, 21 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 12, 21 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k27LhANd018028 12, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Mar 2006 15:43:10 -0600 (CST) 12, 21 -- Message-ID: {440DFE6E.2010101-at-ti.com} 12, 21 -- Date: Tue, 07 Mar 2006 15:43:10 -0600 12, 21 -- From: Becky Holdford {r-holdford-at-ti.com} 12, 21 -- Organization: SC Packaging Development -- FA Development 12, 21 -- User-Agent: Mozilla Thunderbird 0.8 (Windows/20040913) 12, 21 -- X-Accept-Language: en-us, en 12, 21 -- MIME-Version: 1.0 12, 21 -- To: MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 12, 21 -- Subject: 2nd call for papers for Spring meeting of the Texas Society for Microscopy 12, 21 -- Content-Type: text/plain; charset=windows-1252; format=flowed 12, 21 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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Email: camiller-at-anatomy.iupui.edu Name: Caroline A. Miller
Organization: Indiana University, Indiana Microscopy Society
Title-Subject: [Filtered] Our upcoming Spring Meeting, March 20th, 2006
Question: The Spring Meeting of the Indiana Microscopy Society will be Monday, March 20 at the University of Notre Dame, South Bend, IN 8 AM until 3:15 PM in the McKenna Hall Conference Center
Guest Speakers: Daphna Yaniv, PhD, Electron Microscopy Sciences/Quantomix ìEM of Fully Hydrated Samplesî
Eva Chi, PhD, University of Chicago ìRole of Cell Membrane in the Pathogenisis of Alzheimerís Diseaseî
Alex Kandel, PhD, University of Notre Dame ìStructure and Dynamics of Organic Molecules on Surfaces, One Molecule at a Timeî
There will be a best Micrograph and Student Poster Competition A Tour of the Notre Dame Campus will be offered at 3:15 Registration is $10 for members, $20 for non-members Students are free with membership
Breakfast and Lunch Provided For registration and more information go to: indianamicroscopy.org
A very good day to all of you! I apologise that this might be out of place. I am doing some UV-Vis absorption test on my powders in different solvent. I am intending to use carbon tetrachloride or chloroform with a quartz cuvette. However, I am worry that the solvent might damage the lining/joining part of the quartz cuvette. I was informed that strong acid will damage it but I am not too sure CCl4 or chloroform.
I would kindly seek advise from you regarding this. Thank you so much in advance first!!
Have a nice day ahead!
Cheers, YY School of Materials Science and Engineering Nanyang Technological University Singapore
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==============================Original Headers============================== 7, 18 -- From one_twinklestar-at-yahoo.com.sg Tue Mar 7 18:47:47 2006 7, 18 -- Received: from web50006.mail.yahoo.com (web50006.mail.yahoo.com [206.190.38.21]) 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k280lkHm011734 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 18:47:46 -0600 7, 18 -- Received: (qmail 4555 invoked by uid 60001); 8 Mar 2006 00:47:45 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com.sg; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=PhDz60Edu3BvwxWyY+MwzR9o7q7TrMoXIWIFKaqnRTmP64hZzF0dBNFXJS4ocu0rU9Rb6skVASz5goyEUUIxemAXOxr+lHrMk03GGuAdvKACiCU5CuIf0ApYQhlCpFf6eBTqvLxKUCLqp+BDQWX15qow0Vb3OzoTLwoRUH4hnp4= ; 7, 18 -- Message-ID: {20060308004745.4553.qmail-at-web50006.mail.yahoo.com} 7, 18 -- Received: from [202.21.158.12] by web50006.mail.yahoo.com via HTTP; Wed, 08 Mar 2006 08:47:45 CST 7, 18 -- Date: Wed, 8 Mar 2006 08:47:45 +0800 (CST) 7, 18 -- From: Tay Yee Yan {one_twinklestar-at-yahoo.com.sg} 7, 18 -- Subject: Regarding Carbon TetraChloride or Chloroform with Quart Cuvette 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; charset=iso-8859-1 7, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
It's a few decades since I've used such things, but I'm fairly sure they are one-piece, ie just quartz, with no liners or joints. I doubt that strong acid, apart from HF, of course, will damage them either. I used to routinely clean them with the so-called chromic acid (sodium dichromate dissolved in concentrated sulfuric acid) with no problems at all.
cheers
rtch
On 7 Mar 2006 at 19:02, one_twinklestar-at-yahoo.com.sg wrote:
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-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 10, 27 -- From r.sims-at-auckland.ac.nz Tue Mar 7 19:41:23 2006 10, 27 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) 10, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k281fLnO024295 10, 27 -- for {microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 19:41:21 -0600 10, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 10, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 04F693435F; 10, 27 -- Wed, 8 Mar 2006 14:41:20 +1300 (NZDT) 10, 27 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) 10, 27 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 27 -- with ESMTP id 10821-05; Wed, 8 Mar 2006 14:41:19 +1300 (NZDT) 10, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id DAA7234118; 10, 27 -- Wed, 8 Mar 2006 14:41:19 +1300 (NZDT) 10, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 27 -- To: one_twinklestar-at-yahoo.com.sg 10, 27 -- Date: Wed, 08 Mar 2006 14:44:43 +1300 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] Regarding Carbon TetraChloride or Chloroform with Quart Cuvette 10, 27 -- Cc: microscopy-at-microscopy.com 10, 27 -- Message-ID: {440EEDDB.5137.1427CB3-at-localhost} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200603080102.k2812K8Q015619-at-ns.microscopy.com} 10, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 10, 27 -- Content-type: text/plain; charset=US-ASCII 10, 27 -- Content-transfer-encoding: 7BIT 10, 27 -- Content-description: Mail message body 10, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (nomy_nay-at-hotmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, March 7, 2006 at 22:09:53 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both nomy_nay-at-hotmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: In electron microscopy, the higher the voltage the greater the penetrating abilityof the electron beam, but the trade is a reduction of what?
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Email: sue.tyler-at-noaa.gov Name: Sue Tyler
Organization: NOAA
Title-Subject: [Filtered] Pafaffin block storage
Question: Our facility is in the process of building a new paraffin block storage building. The question is, are there specific temperature ranges to maintain optimal block preservation?
Our outside temps. in Maryland are as low as 0 degrees C in the winter.
I have searched the histonet archives but did not come up with any specific temp. Perhaps someone could offer some suggestions. Thanks
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Email: twigg-at-estd.nrl.navy.mil Name: Mark Twigg
Organization: Naval Research Laboratory
Title-Subject: [Filtered] Disposing of Dark Room Chemicals
Question: Lately we have been asked to review our procedures for disposing of chemicals. I was wondering if Kodak has any official recommendations for disposing of D-19 developing and Kodak fixer for TEM negatives.
Higher voltage may 1) Damage your samples easily. For Al foil, you can easily see the beam damage at 300kv 2) Cause multibeam effect when you use the diffraction contrast techniques. Short wavelength means flat Ewards sphere, or more beams are excited.
OF course, cost and maintenance are also problems.
---- Original message ---- } Date: Wed, 8 Mar 2006 08:59:51 -0600 } From: nomy_nay-at-hotmail.com } Subject: [Microscopy] AskAMicroscopist: higher voltage decreases what? } To: clei-at-uiuc.edu } } } } } ------------------------------------------------------------- --------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 5, 27 -- From clei-at-uiuc.edu Wed Mar 8 09:15:19 2006 5, 27 -- Received: from expredir6.cites.uiuc.edu (expredir6.cites.uiuc.edu [128.174.5.97]) 5, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28FFH7K026178 5, 27 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:15:18 -0600 5, 27 -- Received: from expms6.cites.uiuc.edu (expms6.cites.uiuc.edu [128.174.5.43]) 5, 27 -- by expredir6.cites.uiuc.edu (8.12.11/8.12.11) with ESMTP id k28FEpjX000027; 5, 27 -- Wed, 8 Mar 2006 09:14:51 -0600 (CST) 5, 27 -- Received: from expms6.cites.uiuc.edu (localhost.cites.uiuc.edu [127.0.0.1]) 5, 27 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 5, 27 -- with ESMTP id BFR34654; 5, 27 -- Wed, 8 Mar 2006 09:15:12 -0600 (CST) 5, 27 -- Received: from 128.174.5.212 5, 27 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 5, 27 -- with HTTP/1.1; 5, 27 -- Wed, 8 Mar 2006 08:15:12 -0700 5, 27 -- Date: Wed, 8 Mar 2006 08:15:12 -0700 5, 27 -- From: Changhui LEI {clei-at-uiuc.edu} 5, 27 -- Subject: Re: [Microscopy] AskAMicroscopist: higher 5, 27 -- voltage decreases what? 5, 27 -- To: nomy_nay-at-hotmail.com 5, 27 -- Cc: microscopy-at-microscopy.com 5, 27 -- Reply-To: clei-at-uiuc.edu 5, 27 -- X-Mailer: Webmail Mirapoint Direct 3.4.8-GR 5, 27 -- MIME-Version: 1.0 5, 27 -- Message-Id: {1c819e53.95d6f358.81b8000-at-expms6.cites.uiuc.edu} 5, 27 -- Content-Type: text/plain; charset=us-ascii 5, 27 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Mark, You probably need to check your regional/institutional requirements. Here in NYC we are required to use a silver capture system to filter our photographic fix solutions. If we do that, then the D-19 can do down the drain with a lot of water. If we don't use a silver capture system, then we must collect all solutions (developer, stop and fix) and have our Environmental & Life Safety officers take it all away for disposal. If you have a low volume, you can get a simple gravity filtration system that you just pour the fix through when its exhausted. If you have an automated processor, there are traps that can be connected in series with the rest of the system to capture the silver out of the fix. We have both types of systems here, and they work well. The outside contractor comes through periodically to monitor them and change the active capture filter when needed. Its actually pretty painless, and not too expensive. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 21 -- From lcgould-at-med.cornell.edu Wed Mar 8 09:30:22 2006 1, 21 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28FUKbZ031038 1, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:30:20 -0600 1, 21 -- Received: from mpx2.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 21 -- by smtp-gw2.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k28FUHRL028004 1, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:30:18 -0500 (EST) 1, 21 -- Received: from [140.251.48.23] by mpx2.med.cornell.edu 1, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 21 -- with ESMTP id {0IVT00BZDF2HX5A0-at-mpx2.med.cornell.edu} for 1, 21 -- microscopy-at-microscopy.com; Wed, 08 Mar 2006 10:30:17 -0500 (EST) 1, 21 -- Date: Wed, 08 Mar 2006 10:26:02 -0500 1, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 21 -- Subject: Re: [Microscopy] viaWWW: Disposing of Dark Room Chemicals 1, 21 -- In-reply-to: {200603081520.k28FKv8w028026-at-ns.microscopy.com} 1, 21 -- To: twigg-at-estd.nrl.navy.mil, Microscopy Listserver {microscopy-at-microscopy.com} 1, 21 -- Message-id: {p06200709c034a61c8ce8-at-[140.251.48.23]} 1, 21 -- MIME-version: 1.0 1, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 21 -- References: {200603081520.k28FKv8w028026-at-ns.microscopy.com} 1, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.03.08.065107 ==============================End of - Headers==============================
I have some basic very technical questions. I want to prepare MCF-7, HepG2 and Caco-2 cells for TEM observation. My protocol involves the following: - grow cells on 6W multiwell plates up to 70-80% confluency - Wash cells 1x with cold PBS - Add 1 ml cold PBS and detach cells with a cell scraper - Collect the cells in a 1.5 ml eppendorf tube, rince the well 1x with 500µl PBS and merge the volumes ----| total 1.5 ml - Centrifuge at 4°C for 5 min at 1500 RPM - Pipet out carefully the supernatant and carefully add cold Karnovsky fixative - ….
When I follow this protocol with these cells, only the HepG2 give a nice pellet, the other give a too small pellet, Please could you share with me your opinion about this protocol? - Should I forget 6W wells and grow cells on normal petri dishes? I would like to avoid that because I plan to prepare 24 conditions in parallel and working with 24 petri dishes will be a pain. - Is the centrifugation sufficient? Should I increase the centrifugation speed? - Do you have a working protocol for the preparation of these cells for morphological observation? Actually growing these cells in a way that they arrive at the same confluency at the same time is a real challenge since their growth rate are very different. This means that at least one cell line won’t be at optimal confluency at the time of the experiment. I know it’s a question of experience, but I don’t want to want 1 month before starting this experiment :-)
Another question: in parallel to this protocol, I am trying to develop a protocol for the embedding of cell monolayers. The first attempt was not too bad, the embedding basically worked, but when I try to detach the Epon resin from the bottom of the petri dish (I cutted a square with a saw), the surface of the resin is not flat. In addition I don’t know where to cut my pyramid since I don’t see where the cells are on the resin surface. Any clue?
Thank you in advance,
Stephane
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==============================Original Headers============================== 4, 18 -- From nizets2-at-yahoo.com Wed Mar 8 09:39:20 2006 4, 18 -- Received: from web37408.mail.mud.yahoo.com (web37408.mail.mud.yahoo.com [209.191.87.61]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28FdJEq001920 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:39:20 -0600 4, 18 -- Received: (qmail 58964 invoked by uid 60001); 8 Mar 2006 15:39:19 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.com; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 4, 18 -- b=lXojiS18reQ7VfhwrGChK+qQJQofBqFpgp/cSONNcQy2bC84v0/AoWGPKo1xcDx/gMrYHPQWxBDJEWLm3VNKtKpDE+bdVglPv2ioU2n9JFt+kEOxwZOc8SZsqyApi8Kww4Xs9H10q48DO/xeyLlUc9Qk8HMsJ3x6uD+DRefo2+o= ; 4, 18 -- Message-ID: {20060308153919.58962.qmail-at-web37408.mail.mud.yahoo.com} 4, 18 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 07:39:19 PST 4, 18 -- Date: Wed, 8 Mar 2006 07:39:19 -0800 (PST) 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 18 -- Subject: protocol to prepare cells in culture for TEM 4, 18 -- To: microscopy-at-microscopy.com 4, 18 -- MIME-Version: 1.0 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 4, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Stephane, I think my protocol will help you with both of your problems. You certainly can continue to use the 6-well plates. I get samples like that quite frequently. Your fixation and dehydrations can stay the same. For years, I have used a hybrid Epon-analog resin to embed in culture dishes. I use a standard Epon formula but use the followoing compnents:
LX-112 and DMP-30 from Ladd Research Industries DDSA and NMA from Electron Microscopy Sciences
I know it seems weird, but years ago I tried all sorts things, from the "straight" formulations from each vendor to a bunch of mixtures. This one has never reacted with the plastic of the dishes.
Here is how I do the actual embedding of the cell monolayers in the dishes:
After the last 100% ethanol, I remove the alcohol and cover the bottom of the well with a layer of the resin mixture that is about 2 mm deep. I then insert embedding tubes that I've made by cutting the pyramidal bottoms off of BEEM capsules (just slice them with a fresh razor blade and be sure to insert them so that the manufactured end rather than the cut one is sitting against the dish). After I insert labels into the tubes, I put these into the oven at 60 deg. overnight. In the morning, I fill JUST the embedding tubes and return everything to the oven again to finish polymerizing. When the resin is cured, you can grab the tubes with pair of needle-nosed pliers and snap them out. Sometimes a bit of the bottom of the dish comes away with the block, but often you get a very smooth block face. If some of the dish comes up, it is easy to see under a dissecting 'scope and it comes away easily when you trim you block face. I often cut away part of the block face either to keep in reserve or to re-embed to get cross sections, then trim the rest into a narrow rectangle. When you section the resulting block en face, start at 0.25 micrometers (no thicks!), and pick up and Tol. Blue the sections as you get them. You should be able to get smooth thins within a micron or so. I usually trim a very long rectangle and then start to section in such a way that I am a few degrees off of being perfectly en face from top to bottom so that I first get sections from one edge of the rectangle and then have a lot of "acreage" to work through if I need more sections later on.
Disclaimer: I have no financial interest in either Ladd or EMS...I'm just a happy customer who believes in using what works.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 7, 21 -- From lcgould-at-med.cornell.edu Wed Mar 8 10:39:26 2006 7, 21 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28GdNd1023966 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:39:25 -0600 7, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 7, 21 -- by smtp-gw1.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k28GdKbq006737 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 11:39:20 -0500 (EST) 7, 21 -- Received: from [140.251.48.23] by mpx1.med.cornell.edu 7, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 7, 21 -- with ESMTP id {0IVT00D2DI9KVU50-at-mpx1.med.cornell.edu} for 7, 21 -- microscopy-at-microscopy.com; Wed, 08 Mar 2006 11:39:20 -0500 (EST) 7, 21 -- Date: Wed, 08 Mar 2006 11:35:05 -0500 7, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 7, 21 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 7, 21 -- In-reply-to: {200603081559.k28Fxc9f009527-at-ns.microscopy.com} 7, 21 -- To: nizets2-at-yahoo.com, Microscopy Listserver {microscopy-at-microscopy.com} 7, 21 -- Message-id: {p06200701c034b2185c05-at-[140.251.48.23]} 7, 21 -- MIME-version: 1.0 7, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 7, 21 -- References: {200603081559.k28Fxc9f009527-at-ns.microscopy.com} 7, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.03.08.081106 ==============================End of - Headers==============================
I think that if you scrape the cells before fixation you will have a bunch of ripped up cells that have spilled their guts all over the place. We do not scrape until after osmium. If the pellet is very small I do not resuspend it, but I do centrifuge between steps. I have worked with many an invisible pellet.
nizets2-at-yahoo.com wrote:
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Tempting to say simply that the trade is a reduction of contrast. That answer would apply if you were asking about the trade of switching from say 40KV to 100KV on the same microscope.
You can compensate for this by using a smaller objective aperture.
If you are comparing a standard 40 to 100 KV microscope with a high voltage mocroscope then the answer is less straightforward and it is not necessarily so that you have a serious contrast reduction. Perhaps an electron microscope manufacturer will see this question and give you more specific answers.
Best wishes,
Ted Dunn The EMscope Company Ltd. --- nomy_nay-at-hotmail.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question was submitted to Ask-A-Microscopist by } (nomy_nay-at-hotmail.com) } from } http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } on Tuesday, March 7, 2006 at 22:09:53 } Remember to consider the Grade/Age of the student } when considering the Question } --------------------------------------------------------------------------- } Please reply to both nomy_nay-at-hotmail.com as well } as to the Microscopy Listserver } --------------------------------------------------------------------------- } } Email: nomy_nay-at-hotmail.com } Name: Naomi Piyaratna } } Organization: Wollongong University, Australia } } Education: Undergraduate College } } Location: Wollongong, NSW, AUSTRALIA } } Title: Electron Miscroscopy. } } Question: In electron microscopy, the higher the } voltage the greater the penetrating abilityof the } electron beam, but the trade is a reduction of what? } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 8, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Mar } 8 08:40:26 2006 } 8, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 8, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k28EeP1i016821 } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 } Mar 2006 08:40:26 -0600 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } (Unverified) } 8, 12 -- Message-Id: } {p0611040bc0349d3016f2-at-[206.69.208.22]} } 8, 12 -- Date: Wed, 8 Mar 2006 08:40:25 -0600 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: nomy_nay-at-hotmail.com (by way of } Ask-A-Microscopist) } 8, 12 -- Subject: AskAMicroscopist: higher voltage } decreases what? } 8, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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A higher voltage will also reduce amplitude contrast (see below) because the nuclei of the specimen atoms will scatter higher energy electrons less than lower energy electrons. It's quite common for biologists to use a 60kv electron beam routinely to enhance contrast for instance whereas other users may favour 80kv or more for the brighter higher resolution image.
So increasing the voltage seems to produce the same effect as using a larger objective aperture (which will also reduce contrast).
NB as this is an off-list question, I think I should clarify that there are three main types of contrast seen in the transmission electron microscope (unless someone wants to add some more) - amplitude, phase and diffraction contrasts. Amplitude contrast is particularly important up to about 50,000x magnification and heavier elements in the sample with greater nuclear mass will appear darker than lighter elements because of their ability to scatter electrons out of the electron beam.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: clei-at-uiuc.edu
On Mar 8, 2006, at 6:41 AM, nomy_nay-at-hotmail.com wrote:
} Question: In electron microscopy, the higher the voltage the greater } the penetrating abilityof the electron beam, but the trade is a } reduction of what? } Dear Naomi, Contrast. The scattering cross section decreases as electron energy increases up to about 800-1000 kV (depending on the atomic number of the material). This means that for a specific scattered fraction of incident electrons, the allowed thickness will be greater at higher energy; however, since for a given thickness the scattered fraction is smaller, the difference between what happens when the beam strikes your specimen and when it passes through a hole will be less, so there is less contrast. There is no free lunch. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Mar 8 11:57:27 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28HvPKq019527 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 11:57:25 -0600 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 943F43444E 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 09:57:24 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id CCD0633FA9 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 09:57:23 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603081441.k28EfkHo017106-at-ns.microscopy.com} 4, 22 -- References: {200603081441.k28EfkHo017106-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {38e56bb01f1c52c5e20a0103e80d7ec6-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] AskAMicroscopist: higher voltage decreases what? 4, 22 -- Date: Wed, 8 Mar 2006 10:05:58 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Stephanie- There is a protocol for embedding cultured cell layers on our Center for Microscopy & Microanalysis web site. You should skip the step of washing with PBS; it is hard to make this reproducible. Just throw off the medium and pour on the fixative. You recover the cells in what is basically a cast of the cell culture. Ou should get a smooth, glass-like surface where the epoxy resin made contact with the cells. It is hard to cut out the little pieces, but MUCH easier than centrifuging cells down after every step. Antoher advantage is that you keep the relationship of the cells with one another and can see the cell layers (if any), intercellular junctions, etc.
The center web site should appear at the signature line.
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The higher the voltage, the lower the contrast. For ultra-thin sections of biological material a voltage of 60-80 keV is best.
Stephane
--- nomy_nay-at-hotmail.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question was submitted to Ask-A-Microscopist by } (nomy_nay-at-hotmail.com) } from } http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } on Tuesday, March 7, 2006 at 22:09:53 } Remember to consider the Grade/Age of the student } when considering the Question } --------------------------------------------------------------------------- } Please reply to both nomy_nay-at-hotmail.com as well } as to the Microscopy Listserver } --------------------------------------------------------------------------- } } Email: nomy_nay-at-hotmail.com } Name: Naomi Piyaratna } } Organization: Wollongong University, Australia } } Education: Undergraduate College } } Location: Wollongong, NSW, AUSTRALIA } } Title: Electron Miscroscopy. } } Question: In electron microscopy, the higher the } voltage the greater the penetrating abilityof the } electron beam, but the trade is a reduction of what? } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 8, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Mar } 8 08:40:26 2006 } 8, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 8, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k28EeP1i016821 } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 } Mar 2006 08:40:26 -0600 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } (Unverified) } 8, 12 -- Message-Id: } {p0611040bc0349d3016f2-at-[206.69.208.22]} } 8, 12 -- Date: Wed, 8 Mar 2006 08:40:25 -0600 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: nomy_nay-at-hotmail.com (by way of } Ask-A-Microscopist) } 8, 12 -- Subject: AskAMicroscopist: higher voltage } decreases what? } 8, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 7, 20 -- From nizets2-at-yahoo.com Wed Mar 8 11:30:43 2006 7, 20 -- Received: from web37404.mail.mud.yahoo.com (web37404.mail.mud.yahoo.com [209.191.87.57]) 7, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28HUYrV009464 7, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 11:30:35 -0600 7, 20 -- Received: (qmail 68044 invoked by uid 60001); 8 Mar 2006 17:02:30 -0000 7, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 20 -- s=s1024; d=yahoo.com; 7, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 20 -- b=PShnOhKIJY78JybHM2/w+l8sinFMsUxsKDTKohPBgoXeZU+rwzqvRU6vXUYf+jGrPARL/y3UDMKQ4T/ZNCoJLw6rIBR/vdumfn3A1wR30+ha5vOLnaL3CB/my9qlOjD8ZNTpXDoVusBtGh570vupwwCltrkfv8qtKgJfg22xQqo= ; 7, 20 -- Message-ID: {20060308170230.68042.qmail-at-web37404.mail.mud.yahoo.com} 7, 20 -- Received: from [80.122.101.102] by web37404.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 09:02:30 PST 7, 20 -- Date: Wed, 8 Mar 2006 09:02:30 -0800 (PST) 7, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: higher voltage decreases what? 7, 20 -- To: nomy_nay-at-hotmail.com 7, 20 -- Cc: microscopy-at-microscopy.com 7, 20 -- In-Reply-To: {200603081633.k28GXqOZ021992-at-ns.microscopy.com} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; charset=iso-8859-1 7, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Dear Naomi, I don't know how many others have replied to your inquiry. The larger beam-sample interaction volume that results from a higher beam voltage results in the signal coming from deeper in the sample, rather than just from the surface. This gives information from deeper in the sample, but sacrifices information from the very surface. If you need to see small features on the surface of your sample, a lower accelerating voltage is better. I hope this helps. Any basic SEM text should cover this point. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- X-from: {nomy_nay-at-hotmail.com} To: {mager-at-interchange.ubc.ca} Sent: Wednesday, March 08, 2006 7:57 AM
The ongoing discussion about contrast brings to mind another question. If one wants to add enough heavy metal to label a singular structure on a biological tissue thin section, how much metal is required to obtain a useful signal on a standard TEM? Would a STEM system allow one to "see" the structure with a lower amount of heavy metal label? Or does an energy filtered electron microscope (like the Zeiss 902) permit one to resolve smaller clusters?
I remember that some gold-linked antibody probes used fairly small gold clusters (11 atoms perhaps?) to improve penetration into the section, but that these ABs were only made visible after silver enhancement for routine TEM. When does a cluster of metal atoms become resolvable in a minimally stained thin section? -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
==============================Original Headers============================== 4, 21 -- From hall-at-aecom.yu.edu Wed Mar 8 14:06:39 2006 4, 21 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 4, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28K6c8f026310 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 14:06:38 -0600 4, 21 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 4, 21 -- by mailgw.aecom.yu.edu (8.12.11/8.12.11) with SMTP id k28K6WnP016413 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 15:06:38 -0500 4, 21 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 4, 21 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006030815063806598 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 08 Mar 2006 15:06:38 -0500 4, 21 -- Received: from [129.98.90.160] (worm.aecom.yu.edu [129.98.90.160]) 4, 21 -- by post.aecom.yu.edu (Postfix) with ESMTP id EB5852FD5 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 15:06:37 -0500 (EST) 4, 21 -- Mime-Version: 1.0 4, 21 -- X-Sender: hall-at-mailserver.aecom.yu.edu 4, 21 -- Message-Id: {a05100306c034e75199ac-at-[129.98.90.160]} 4, 21 -- Date: Wed, 8 Mar 2006 15:06:35 -0500 4, 21 -- To: microscopy-at-microscopy.com 4, 21 -- From: David Hall {hall-at-aecom.yu.edu} 4, 21 -- Subject: contrast and imaging small clusters of heavy metals 4, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
} Hi Sephanie Can you use glass coverslips? If so } I have a protocol I can send you where you can } embed the whole coverslip (cell side down) in a } chang embedding mold. You then remove the glass } using hydrofluoric acid, punch out small resin } circles of cells using a leather punch . Then } re-attach on to a blank resin stub and } section. That way you can get many blocks from } 1 coverslip. As mentioned earlier, avoid the PBS } step and scraping as both can destroy the morphology.
JoAnn Buchanan Stanford University School of Medicine
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Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
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Greater high voltage in a TEM is one of the few things in nature that does not have a lot of serious "Cons" that outweigh or balance the "Pros." Granted that increased radiation concerns and somewhat less contrast attend increasing the voltage, but on the plus side, the increased penetration, easier specimen preparation, improved resolution, plus others pros are BIG advantages.
Please forgive me if I point out that should you have a radiation sensitive specimen that you can always lower the voltage on a 300keV TEM for that specimen, but you can't raise the voltage on a 100keV machine to allow you to see through a thick specimen.
Ron Anderson
drteddunne-at-yahoo.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello Naomi, } } Tempting to say simply that the trade is a reduction } of contrast. That answer would apply if you were } asking about the trade of switching from say 40KV to } 100KV on the same microscope. } } You can compensate for this by using a smaller } objective aperture. } } If you are comparing a standard 40 to 100 KV } microscope with a high voltage mocroscope then the } answer is less straightforward and it is not } necessarily so that you have a serious contrast } reduction. Perhaps an electron microscope manufacturer } will see this question and give you more specific } answers. } } Best wishes, } } } Ted Dunn } The EMscope Company Ltd. } --- nomy_nay-at-hotmail.com wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The } } Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } This Question was submitted to Ask-A-Microscopist by } } (nomy_nay-at-hotmail.com) } } from } } } } } http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } } } on Tuesday, March 7, 2006 at 22:09:53 } } Remember to consider the Grade/Age of the student } } when considering the Question } } } } } --------------------------------------------------------------------------- } } } Please reply to both nomy_nay-at-hotmail.com as well } } as to the Microscopy Listserver } } } } } --------------------------------------------------------------------------- } } } Email: nomy_nay-at-hotmail.com } } Name: Naomi Piyaratna } } } } Organization: Wollongong University, Australia } } } } Education: Undergraduate College } } } } Location: Wollongong, NSW, AUSTRALIA } } } } Title: Electron Miscroscopy. } } } } Question: In electron microscopy, the higher the } } voltage the greater the penetrating abilityof the } } electron beam, but the trade is a reduction of what? } } } } } } } --------------------------------------------------------------------------- } } } } } ==============================Original } } Headers============================== } } 8, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Mar } } 8 08:40:26 2006 } } 8, 12 -- Received: from [206.69.208.22] } } (mac22.zaluzec.com [206.69.208.22]) } } 8, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } } ESMTP id k28EeP1i016821 } } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 } } Mar 2006 08:40:26 -0600 } } 8, 12 -- Mime-Version: 1.0 } } 8, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } } (Unverified) } } 8, 12 -- Message-Id: } } {p0611040bc0349d3016f2-at-[206.69.208.22]} } } 8, 12 -- Date: Wed, 8 Mar 2006 08:40:25 -0600 } } 8, 12 -- To: microscopy-at-microscopy.com } } 8, 12 -- From: nomy_nay-at-hotmail.com (by way of } } Ask-A-Microscopist) } } 8, 12 -- Subject: AskAMicroscopist: higher voltage } } decreases what? } } 8, 12 -- Content-Type: text/plain; } } charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 10, 20 -- From drteddunne-at-yahoo.com Wed Mar 8 10:47:40 2006 } 10, 20 -- Received: from web33401.mail.mud.yahoo.com (web33401.mail.mud.yahoo.com [68.142.206.133]) } 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28Glche027028 } 10, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:47:39 -0600 } 10, 20 -- Received: (qmail 95631 invoked by uid 60001); 8 Mar 2006 16:47:38 -0000 } 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 10, 20 -- s=s1024; d=yahoo.com; } 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 10, 20 -- b=xpdlSSAAs0maYw04oUmPXHuqrXZmk07hPoZ8sv26sWh9auopxV0y/Sk8pC2kl0jDWpfW2TuOMk+g98SFzYOPojX2MrElhArfxv/3WaIY53wt88EQsbbUfuabYnRZpoWvdipk4Hgzqjz6poJVOSsr+FJE9qaEYUtJotg0YNB7sP8= ; } 10, 20 -- Message-ID: {20060308164738.95629.qmail-at-web33401.mail.mud.yahoo.com} } 10, 20 -- Received: from [202.47.247.156] by web33401.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 08:47:38 PST } 10, 20 -- Date: Wed, 8 Mar 2006 08:47:38 -0800 (PST) } 10, 20 -- From: ted dunn {drteddunne-at-yahoo.com} } 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: higher voltage decreases what? } 10, 20 -- To: nomy_nay-at-hotmail.com } 10, 20 -- Cc: microscopy-at-microscopy.com } 10, 20 -- In-Reply-To: {200603081542.k28FgiZ6003132-at-ns.microscopy.com} } 10, 20 -- MIME-Version: 1.0 } 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 } 10, 20 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 19 -- From randerson20-at-tampabay.rr.com Wed Mar 8 15:12:46 2006 5, 19 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01-smtplb.tampabay.rr.com [65.32.5.131]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28LCjXY012167 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 8 Mar 2006 15:12:46 -0600 5, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 19 -- by ms-smtp-01.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k28HBtjp013043 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 8 Mar 2006 12:11:57 -0500 (EST) 5, 19 -- Message-ID: {440F1059.5060502-at-tampabay.rr.com} 5, 19 -- Date: Wed, 08 Mar 2006 12:11:53 -0500 5, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: higher voltage decreases what? 5, 19 -- References: {200603081650.k28Gofmh028153-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200603081650.k28Gofmh028153-at-ns.microscopy.com} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
First, washing cells and centrifuging them before fixing will cause some changes in the morphology. Usually the changes are hidden after the cells have been embedded in epoxy resin because the resin is so good at holding the damaged cells together. As a general rule, for TEM examination it is best to handle the cells as little as possible, even after fixation. I think there is an old Nature paper by Pernilla et al that clearly demonstrates a loss of protein from unfixed cells during centrifugation.
After all, we don't need to wash cells if we don't have to. We are electron microscopists and can easily face the challenge of differentiating what is in the cells from what is outside. Why wash all the outside stuff away unless it is absolutely necessary?
We process cell monolayers in dishes similar to those you use and when we want to examine a pellet we fix first by adding double strength fixative to the cells as they are growing, and at the temperature they are growing at. After all, it is well known that changing the temperature of the cells can cause significant changes within the cells.
Once the fixative is added we wait 30 sec and then carefully scrape the cells from the substrate. Instead of regular cell scrapers we use small pieces of carefully trimmed teflon, or even shaped orange sticks that have been soaked in buffer before use.
Each dish is fixed and scraped individually and the cells immediately transferred to a tube for pelleting. We use an Eppendorf centrifuge that we bring up to top speed and then immediately let run down. It is important that each dish is treated individually and not batch processed as is routine in most labs. The only problem with a 6-well dish is that such individual attention is not easy when all the culture wells are on the same plate.
Once the fixed cells have been pelleted, we then leave the pellets to cool on ice. At some point we will add fresh, single strength fixative so that the pellet continues fixing. What is interesting is that if you pellet the cells while they are fixing and leave them as a pellet, a very strong clump of cells is formed. The cells do not fall apart so there is no need to continue centrifuging after that one pelleting step, and the pellet can be cut into smaller pieces for easier dehydration, infiltration and embedding.
Your dehydration and embedding protocol is not really important. Different dehydration agents produce different results, and the resin you use will also affect staining and sectioning qualities.
One of the most important parts of the process is the amount of time you grow your cells before fixing them. We always try to let the cells grow for at least 3 days before using them. I know that is not always possible when looking at transient transfections and other short-lived experiments, but letting the cells grow really does make a difference to the morphology. Again, I think there is lots of old (pre-pdf era) literature to dig into on the effect of growth on morphology (more proteins are made by the cells).
I think the processing of monolayers have been nicely covered already by other contributors, but I can add one extra point. If you want to remove glass or Thermaox slides from an epoxy resin block using liquid nitrogen, then the method works best if the resin has not been fully polymerized. Put the blocks in the oven in the morning and try to remove the blocks later in the day when the resin has hardened a little.
Best regards,
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
} From: {nizets2-at-yahoo.com} } Reply-To: {nizets2-at-yahoo.com} } Date: Wed, 8 Mar 2006 11:29:58 -0600 } To: {pwebster-at-hei.org} } Subject: [Microscopy] protocol to prepare cells in culture for TEM } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear colleagues and listers, } } I have some basic very technical questions. I want } to prepare MCF-7, } HepG2 and Caco-2 cells for TEM observation. } My protocol involves the following: } - grow cells on 6W multiwell plates up to } 70-80% confluency } - Wash cells 1x with cold PBS } - Add 1 ml cold PBS and detach cells with a } cell scraper } - Collect the cells in a 1.5 ml eppendorf } tube, rince the } well 1x with 500µl PBS and merge the volumes ----| } total 1.5 ml } - Centrifuge at 4°C for 5 min at 1500 RPM } - Pipet out carefully the supernatant and } carefully add cold } Karnovsky fixative } - ∑. } } When I follow this protocol with these cells, only the } HepG2 give a } nice pellet, the other give a too small pellet, } Please could you share with me your opinion about } this protocol? } - Should I forget 6W wells and grow cells } on normal petri } dishes? I would like to avoid that because I plan to } prepare 24 } conditions in parallel and working with 24 petri } dishes will be a pain. } - Is the centrifugation sufficient? Should } I increase the } centrifugation speed? } - Do you have a working protocol for the } preparation of } these cells for morphological observation? } Actually growing these cells in a way that they } arrive at the same } confluency at the same time is a real challenge since } their growth rate } are very different. This means that at least one cell } line won‚t be at } optimal confluency at the time of the experiment. I } know it‚s a question } of experience, but I don‚t want to want 1 month before } starting this } experiment :-) } } Another question: in parallel to this protocol, I am } trying to } develop a protocol for the embedding of cell } monolayers. The first attempt } was not too bad, the embedding basically worked, but } when I try to detach } the Epon resin from the bottom of the petri dish (I } cutted a square } with a saw), the surface of the resin is not flat. In } addition I don‚t } know where to cut my pyramid since I don‚t see where } the cells are on the } resin surface. Any clue? } } Thank you in advance, } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 4, 18 -- From nizets2-at-yahoo.com Wed Mar 8 09:39:20 2006 } 4, 18 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.87.61]) } 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28FdJEq001920 } 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:39:20 -0600 } 4, 18 -- Received: (qmail 58964 invoked by uid 60001); 8 Mar 2006 15:39:19 } -0000 } 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 4, 18 -- s=s1024; d=yahoo.com; } 4, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-T } ransfer-Encoding; } 4, 18 -- } b=lXojiS18reQ7VfhwrGChK+qQJQofBqFpgp/cSONNcQy2bC84v0/AoWGPKo1xcDx/gMrYHPQWxBDJ } EWLm3VNKtKpDE+bdVglPv2ioU2n9JFt+kEOxwZOc8SZsqyApi8Kww4Xs9H10q48DO/xeyLlUc9Qk8H } MsJ3x6uD+DRefo2+o= ; } 4, 18 -- Message-ID: {20060308153919.58962.qmail-at-web37408.mail.mud.yahoo.com} } 4, 18 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via } HTTP; Wed, 08 Mar 2006 07:39:19 PST } 4, 18 -- Date: Wed, 8 Mar 2006 07:39:19 -0800 (PST) } 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 4, 18 -- Subject: protocol to prepare cells in culture for TEM } 4, 18 -- To: microscopy-at-microscopy.com } 4, 18 -- MIME-Version: 1.0 } 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 4, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers==============================
==============================Original Headers============================== 20, 20 -- From PWebster-at-hei.org Wed Mar 8 16:55:57 2006 20, 20 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 20, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28Mts57009838 20, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 16:55:55 -0600 20, 20 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 20, 20 -- Wed, 8 Mar 2006 22:52:26 +0000 20, 20 -- User-Agent: Microsoft-Entourage/11.2.1.051004 20, 20 -- Date: Wed, 08 Mar 2006 14:55:52 -0800 20, 20 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 20, 20 -- From: "Webster, Paul" {PWebster-at-hei.org} 20, 20 -- To: {nizets2-at-yahoo.com} , {microscopy-at-microscopy.com} 20, 20 -- Message-ID: {C034A0F8.8A9C%PWebster-at-hei.org} 20, 20 -- Thread-Topic: [Microscopy] protocol to prepare cells in culture for TEM 20, 20 -- Thread-Index: AcZDA3MAsXpDM672EdqlqAANk7Zh7g== 20, 20 -- In-Reply-To: {200603081729.k28HTvGp009203-at-ns.microscopy.com} 20, 20 -- Mime-version: 1.0 20, 20 -- Content-type: text/plain; 20, 20 -- charset="UTF-8" 20, 20 -- Content-Transfer-Encoding: 8bit 20, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k28Mts57009838 ==============================End of - Headers==============================
On Mar 8, 2006, at 7:17 AM, clei-at-uiuc.edu wrote:
} Higher voltage may } 1) Damage your samples easily. For Al foil, you can easily } see the beam damage at 300kv } 2) Cause multibeam effect when you use the diffraction } contrast techniques. Short wavelength means flat Ewards } sphere, or more beams are excited. } } OF course, cost and maintenance are also problems. } Dear Changhui & Naomi, All true, and one can take advantage of both. 1) Although the elastic and total cross sections decrease with increasing energy, the inelastic cross section rises with increasing energy, so by using an energy filter or collecting position-tagged spectra--a complete energy spectrum at each pixel obtained with the dose used for imaging--one can increase the contrast by using only the unscattered and elastically scattered electrons to produce the image, and one can collect (almost) all the electrons incident on the specimen and make use of their energy losses to identify the constituents; i.e., do element mapping. 2) Acquiring the higher-order diffraction information will allow one to get higher resolution, and diffraction contrast techniques are not the only case where this is true. I can't speak to the cost problem, but having maintained a 1.2 MeV HVEM for many years, I can say that, while more difficult than just buying a service contract, a dedicated staff can maintain good performance from such an instrument for decades. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Mar 8 17:00:43 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28N0fbw011238 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 17:00:42 -0600 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 72F153456A 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 15:00:41 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id A465235D52 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 15:00:40 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603081517.k28FHOHf026899-at-ns.microscopy.com} 4, 22 -- References: {200603081517.k28FHOHf026899-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {ffad539d04d4f31bc0412cf77c665982-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: higher 4, 22 -- Date: Wed, 8 Mar 2006 15:09:15 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Hi Stephane I would not scrape cells until you have fixed them.
As to flat embedding cells; grow cells on Aclar film (EMS or Pella) fix and osmificate on the film process the aclar film just as you would any other prep embed and bake the plastic the film will still soft and will peal off, leaving a smooth plastic block with the cells in the plastic because the cells are just on the surface, and the block is smooth, and the cells are osmificated, they can easily be seen under a microscope I then take a fine saw and cut rectangular blocks out of the plastic and mount them in the microtome. This works well for X-sections of your cells. If you wish to section in the plane of the film, cut out small pieces and glue them (cell side up) to a blank
If you have any questions, contact me off-list. I probably have a protocol sitting around I could send you. David
On Mar 8, 2006, at 10:05 AM, nizets2-at-yahoo.com wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Dear colleagues and listers, } } I have some basic very technical questions. I want } to prepare MCF-7, } HepG2 and Caco-2 cells for TEM observation. } My protocol involves the following: } - grow cells on 6W multiwell plates up to } 70-80% confluency } - Wash cells 1x with cold PBS } - Add 1 ml cold PBS and detach cells with a } cell scraper } - Collect the cells in a 1.5 ml eppendorf } tube, rince the } well 1x with 500µl PBS and merge the volumes ----| } total 1.5 ml } - Centrifuge at 4°C for 5 min at 1500 RPM } - Pipet out carefully the supernatant and } carefully add cold } Karnovsky fixative } - } . } } When I follow this protocol with these cells, only the } HepG2 give a } nice pellet, the other give a too small pellet, } Please could you share with me your opinion about } this protocol? } - Should I forget 6W wells and grow cells } on normal petri } dishes? I would like to avoid that because I plan to } prepare 24 } conditions in parallel and working with 24 petri } dishes will be a pain. } - Is the centrifugation sufficient? Should } I increase the } centrifugation speed? } - Do you have a working protocol for the } preparation of } these cells for morphological observation? } Actually growing these cells in a way that they } arrive at the same } confluency at the same time is a real challenge since } their growth rate } are very different. This means that at least one cell } line won’t be at } optimal confluency at the time of the experiment. I } know it’s a question } of experience, but I don’t want to want 1 month before } starting this } experiment :-) } } Another question: in parallel to this protocol, I am } trying to } develop a protocol for the embedding of cell } monolayers. The first attempt } was not too bad, the embedding basically worked, but } when I try to detach } the Epon resin from the bottom of the petri dish (I } cutted a square } with a saw), the surface of the resin is not flat. In } addition I don’t } know where to cut my pyramid since I don’t see where } the cells are on the } resin surface. Any clue? } } Thank you in advance, } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 4, 18 -- From nizets2-at-yahoo.com Wed Mar 8 09:39:20 2006 } 4, 18 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.87.61]) } 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id } k28FdJEq001920 } 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:39:20 } -0600 } 4, 18 -- Received: (qmail 58964 invoked by uid 60001); 8 Mar 2006 } 15:39:19 -0000 } 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 4, 18 -- s=s1024; d=yahoo.com; } 4, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type: } Content-Transfer-Encoding; } 4, 18 -- } b=lXojiS18reQ7VfhwrGChK+qQJQofBqFpgp/cSONNcQy2bC84v0/AoWGPKo1xcDx/ } gMrYHPQWxBDJEWLm3VNKtKpDE+bdVglPv2ioU2n9JFt+kEOxwZOc8SZsqyApi8Kww4Xs9H1 } 0q48DO/xeyLlUc9Qk8HMsJ3x6uD+DRefo2+o= ; } 4, 18 -- Message-ID: } {20060308153919.58962.qmail-at-web37408.mail.mud.yahoo.com} } 4, 18 -- Received: from [80.122.101.102] by } web37408.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 07:39:19 PST } 4, 18 -- Date: Wed, 8 Mar 2006 07:39:19 -0800 (PST) } 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 4, 18 -- Subject: protocol to prepare cells in culture for TEM } 4, 18 -- To: microscopy-at-microscopy.com } 4, 18 -- MIME-Version: 1.0 } 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 4, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers==============================
==============================Original Headers============================== 8, 23 -- From Elliott-at-Arizona.edu Wed Mar 8 14:19:52 2006 8, 23 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 8, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28KJpwr030045 8, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2006 14:19:51 -0600 8, 23 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 8, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 8F665D3E079 8, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2006 13:19:50 -0700 (MST) 8, 23 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 8, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 7AC30D3B759 8, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2006 13:19:46 -0700 (MST) 8, 23 -- Mime-Version: 1.0 (Apple Message framework v623) 8, 23 -- In-Reply-To: {200603081705.k28H54vS000881-at-ns.microscopy.com} 8, 23 -- References: {200603081705.k28H54vS000881-at-ns.microscopy.com} 8, 23 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 8, 23 -- Message-Id: {c583c629128f46efca713255cec9301b-at-Arizona.edu} 8, 23 -- From: David Elliott {Elliott-at-Arizona.edu} 8, 23 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 8, 23 -- Date: Wed, 8 Mar 2006 13:19:46 -0700 8, 23 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 8, 23 -- X-Mailer: Apple Mail (2.623) 8, 23 -- X-Virus-Scanned: amavisd-new at email.arizona.edu 8, 23 -- Content-Transfer-Encoding: 8bit 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k28KJpwr030045 ==============================End of - Headers==============================
There is another interesting approach, especially since you are working with c. Elegans. NT-MDT has just announced a new device which integrates an atomic force microscope with a Leica ultramicrotome. All of the early work has been done on either polymers or on c. elegans, especially by Dr. M. Mueller and Dr. N. Matsko at ETH in Zurich. The AFM uses local differences in elasticity and other physical properties to provide contrast, so in many cases, there is no need to stain with heavy metals. Also, because the AFM images from the block face, in sequential planes, it provides perfectly aligned sections for 3D reconstruction.
I understand that a demo unit will be available here in the States sometime around mid-year.
If you are interested in images and/or further information, please contact me off-line
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). For details, call us at 972-943-8011 and ask for Ken Piel.
CAVEAT: MME is involved in the support of this techology and therefore has a financial interest.
At 03:14 PM 3/8/2006, hall-at-aecom.yu.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 14, 18 -- From bfoster-at-mme1.com Wed Mar 8 18:13:30 2006 14, 18 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 14, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k290DRk8029991 14, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 18:13:28 -0600 14, 18 -- Received: (qmail 27665 invoked from network); 8 Mar 2006 18:15:11 -0500 14, 18 -- Received: from host57-99.rancor.birch.net (HELO Barb-XP.mme1.com) (65.17.57.99) 14, 18 -- by enterprise.5starpro.com with SMTP; 8 Mar 2006 18:15:11 -0500 14, 18 -- Message-Id: {7.0.1.0.0.20060308170743.01d8d888-at-mme1.com} 14, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 18 -- Date: Wed, 08 Mar 2006 17:13:22 -0600 14, 18 -- To: hall-at-aecom.yu.edu, microscopy-at-microscopy.com 14, 18 -- From: Barbara Foster {bfoster-at-mme1.com} 14, 18 -- Subject: Re: [Microscopy] contrast and imaging small clusters of heavy 14, 18 -- metals 14, 18 -- In-Reply-To: {200603082114.k28LEj9f012730-at-ns.microscopy.com} 14, 18 -- References: {200603082114.k28LEj9f012730-at-ns.microscopy.com} 14, 18 -- Mime-Version: 1.0 14, 18 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
In the same vein as the complaints about having to go through SPAM filter certifications when sending listserver messages:::::::
Why is it that people do not "unsubscribe" from the listserver when they attend a meeting, go on vacation or do whatever it is they put in their auto-reply messages?
I get enough junk mail already so getting a barrage of these messages after I post on the listserver has one effect only - I don't post messages.
Now I wait for the auto-replies to arrive.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
==============================Original Headers============================== 9, 18 -- From PWebster-at-hei.org Wed Mar 8 18:57:36 2006 9, 18 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k290vY7T010073 9, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 18:57:35 -0600 9, 18 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 9, 18 -- Thu, 9 Mar 2006 00:54:06 +0000 9, 18 -- User-Agent: Microsoft-Entourage/11.2.1.051004 9, 18 -- Date: Wed, 08 Mar 2006 16:57:33 -0800 9, 18 -- Subject: Out-of office replies 9, 18 -- From: "Webster, Paul" {PWebster-at-hei.org} 9, 18 -- To: {microscopy-at-microscopy.com} 9, 18 -- Message-ID: {C034BD7D.8AC0%PWebster-at-hei.org} 9, 18 -- Thread-Topic: Out-of office replies 9, 18 -- Thread-Index: AcZDFHK8sUqIzK8HEdqlqAANk7Zh7g== 9, 18 -- Mime-version: 1.0 9, 18 -- Content-type: text/plain; 9, 18 -- charset="US-ASCII" 9, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
This is most likely an Outlook issue. I use Eudora and simply set up an automatic kill filter for that subject (hence I changed this message's subject or I would not see my own posting--actually a good test). Your message's Subject does not exactly match Outlook's format so it did not get killed at my end.
Nestor has made numerous postings/Administrivia about this. I guess the problem still continues for those who don't have the ability to do filtering.
gary g.
At 05:42 PM 3/8/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Wed Mar 8 20:22:04 2006 9, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k292M3Y5001772 9, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 20:22:03 -0600 9, 20 -- Received: (qmail 11897 invoked from network); 8 Mar 2006 18:22:02 -0800 9, 20 -- Received: by simscan 1.1.0 ppid: 11893, pid: 11894, t: 0.1657s 9, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 20 -- by qsmtp1 with SMTP; 8 Mar 2006 18:22:02 -0800 9, 20 -- Message-Id: {6.2.3.4.2.20060308181546.02038dd0-at-mail.calweb.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 9, 20 -- Date: Wed, 08 Mar 2006 18:22:02 -0800 9, 20 -- To: PWebster-at-hei.org 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] Outofoffice replies 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200603090142.k291gfbj024539-at-ns.microscopy.com} 9, 20 -- References: {200603090142.k291gfbj024539-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I am been dismayed by the number of responses that answer the question as if it is for TEM.
Maybe I am missing something, but TEM is not mentioned in the question. Neither is the voltage range, such as 80KeV or 300KeV.
Hence, it is not obvious if the question relates to TEM, SEM, etc.....
My knee-jerk is response is: high penetration --- high transmission less reflection --- less surface sensitive
This would work equally well for SEM, STEM, TEM, AEM, etc......
JQuinn
PS: Neither is 'bio' vs 'materials'.
--------------------------------------------------------------------------- } } Email: nomy_nay-at-hotmail.com } Name: Naomi Piyaratna } } Organization: Wollongong University, Australia } } Education: Undergraduate College } } Location: Wollongong, NSW, AUSTRALIA } } Title: Electron Miscroscopy. } } Question: In electron microscopy, the higher the } voltage the greater the penetrating abilityof the } electron beam, but the trade is a reduction of what? } }
==============================Original Headers============================== 11, 12 -- From jquinn-at-www.matscieng.sunysb.edu Wed Mar 8 13:07:04 2006 11, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 11, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28J6wCH009419 11, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 13:07:01 -0600 11, 12 -- Received: (from jquinn-at-localhost) 11, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id k28J5Q113161 11, 12 -- for microscopy-at-microscopy.com; Wed, 8 Mar 2006 14:05:26 -0500 11, 12 -- Date: Wed, 8 Mar 2006 14:05:26 -0500 11, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 11, 12 -- Message-Id: {200603081905.k28J5Q113161-at-www.matscieng.sunysb.edu} 11, 12 -- To: microscopy-at-microscopy.com 11, 12 -- Subject: re: higher voltage ==============================End of - Headers==============================
Warm thanks to everyone for taking the time to instruct me about their protocol. I received a lot of answers, and lots of great ideas. Now I have to make a choice ;-)
Apparently general congruence can be observed for important steps: - Never wash with PBS, prefer direct fixation - Avoid scraping live cells. For this point one can ask why cell scrapers exist if they are so damaging to live cells. - There is still a possibility to use propyleneoxide in petri dishes, though it requires practice. I will keep this possibility as a "last resource" if nothing else works ;-) - Detaching the resin from the support after 12h (before complete curing) helps. - Growing cells in 6W format is definitely possible ;-) (which is a great new)
Some great ideas I will follow: - Using cut BEEM capsules during embedding of monolayers - Carbon-coating glass slides (I mean this one is really great isn't it?) - Fixing cells in the medium for a short time, collecting and centrifuging, then continuing fixation on the pellet (it helps a lot since I abandoned in situ fixation because I had a loose pellet and then too few cells per section) - When processing monolayers, minimize the time of contact with "extracting" substance (dehydration)
I don't always need to know the orientation of the cells, and so it was great to receive ideas for both pellet and monolayer embedding.
P.S1: I got only one clue how to localize the cells after monolayer embedding. Other help would still be welcome.
P.S2: Stephane is a french name, and it is different from Stephanie :D
Finally, I wish good luck to Pat in the land of the Sauerkraut and the biggest beer drinkers of the world.
Warm regards,
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 12, 18 -- From nizets2-at-yahoo.com Thu Mar 9 02:37:47 2006 12, 18 -- Received: from web37404.mail.mud.yahoo.com (web37404.mail.mud.yahoo.com [209.191.87.57]) 12, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k298bks7027101 12, 18 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 02:37:47 -0600 12, 18 -- Received: (qmail 10130 invoked by uid 60001); 9 Mar 2006 08:37:39 -0000 12, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 12, 18 -- s=s1024; d=yahoo.com; 12, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 12, 18 -- b=LeDk2C4mCnIKyAhulaQt4TsoXvwyskvmHvmvfxQlCue6Kr4RWQnnRl9ewssQWsbst3NrCv1WSRELoVmvUOlUwR1TqzLi0vU/XDYiqTrtH/2o5JLqcBV2RJjn4eT9GHa7EwEIJ4/evAkT5gSsbgCH4qtIfK6ydw3YbrOg9WjDUD0= ; 12, 18 -- Message-ID: {20060309083739.10128.qmail-at-web37404.mail.mud.yahoo.com} 12, 18 -- Received: from [80.122.101.102] by web37404.mail.mud.yahoo.com via HTTP; Thu, 09 Mar 2006 00:37:39 PST 12, 18 -- Date: Thu, 9 Mar 2006 00:37:39 -0800 (PST) 12, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 12, 18 -- Subject: processing cell in culture for TEM: summary 12, 18 -- To: microscopy-at-microscopy.com 12, 18 -- MIME-Version: 1.0 12, 18 -- Content-Type: text/plain; charset=iso-8859-1 12, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Sorry if I digress a bit, but I am new to the field of EDX. I thought that higher voltages gave a higher signal in EDX, and so a higher sensitivity. Is it not true?
Stéphane
http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi, } } In SEM, you will lose surface sensitivity. } } In TEM, scattering cross sections decrease which is } not good for EDX and EELS. } } Hongqi } } Dept. of Materials Science and Engineering } Pennsylvania State University } University Park, PA 16802 } email: hud105-at-psu.edu } } } ==============================Original
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I wish to thank the many members of this list who replied to my rather poorly posed question about EM costs. All of the replies were informative and helpful. They came in two flavors. Some wrote to say that I did not provide enough information to make even a rough guess, but they were kind enough to list the kinds of things I needed to specify or know in order to make cost estimates for these labs. Others described their labs and provided some cost estimates for their setups. I was also referred to some useful articles. I also had some replies from companies that sell new or used equipment, with information and prices. All told, I had about 2 dozen replies and followups in the two weeks following the initial request.
--aryeh -- Aryeh Weiss School of Engineering Bar Ilan University Ramat Gan 52900 Israel
Ph: 972-3-5317638 FAX: 972-3-5340697
==============================Original Headers============================== 4, 29 -- From aryeh-at-cc.huji.ac.il Thu Mar 9 04:09:16 2006 4, 29 -- Received: from tamar.os.biu.ac.il (tamar.os.biu.ac.il [132.70.60.24]) 4, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29A9FBQ015738 4, 29 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 04:09:15 -0600 4, 29 -- Received: from ismss-1.biu.ac.il (ismss.biu.ac.il [132.70.84.150]) 4, 29 -- by tamar.os.biu.ac.il (8.12.11/8.12.11/BIU) with ESMTP id k29A99KL696542 4, 29 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 12:09:13 +0200 4, 29 -- Received: from cc.huji.ac.il ([132.70.133.31]RDNS failed) by 4, 29 -- ismss-1.biu.ac.il with InterScan Message Security Suite; Thu, 09 Mar 2006 4, 29 -- 12:10:40 +0200 4, 29 -- Message-ID: {440FFEBF.40509-at-cc.huji.ac.il} 4, 29 -- Date: Thu, 09 Mar 2006 12:09:03 +0200 4, 29 -- From: Aryeh Weiss {aryeh-at-cc.huji.ac.il} 4, 29 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) 4, 29 -- Gecko/20030624 Netscape/7.1 (ax) 4, 29 -- X-Accept-Language: en-us, en, he 4, 29 -- MIME-Version: 1.0 4, 29 -- To: microscopy-at-microscopy.com 4, 29 -- Subject: cost of TEM, SEM and AFM 4, 29 -- Content-Type: text/plain; 4, 29 -- charset=us-ascii; 4, 29 -- format=flowed 4, 29 -- Content-Transfer-Encoding: 7bit 4, 29 -- X-imss-version: 2.038 4, 29 -- X-imss-result: Passed 4, 29 -- X-imss-scanInfo: M:P L:E SM:0 4, 29 -- X-imss-tmaseResult: TT:0 TS:0.0000 TC:00 TRN:0 TV:3.52.1006(14312.002) 4, 29 -- X-imss-scores: Clean:23.91834 C:2 M:3 S:5 R:5 4, 29 -- X-imss-settings: Baseline:2 C:1 M:1 S:1 R:1 (0.1500 0.1500) ==============================End of - Headers==============================
We are selling our practically unused cryo system for SEM, Oxford CT 1500B, bought in 1996, because of lack of space and suitable projects. The price is set to 5000 USD, which is far below the price of a new comparable system. The buyer will have to pay for the transportation. Please, ask for more information, if you are interested!
****************************** Kerstin Brismar B.Sc., Research engineer, Photographer Dept. of Crop Science SLU (Swedish University of Agricultural Sciences) P.O. Box 44 (Delivery: Växtskyddsvägen 1) SE-230 53 Alnarp, Sweden Phone: +46 40 41 55 05 Fax: +46 40 41 55 19 E-mail: Kerstin.Brismar-at-vv.slu.se ******************************
==============================Original Headers============================== 6, 17 -- From Kerstin.Brismar-at-vv.slu.se Thu Mar 9 04:33:27 2006 6, 17 -- Received: from alnus.slu.se (alnus.slu.se [194.47.49.5]) 6, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29AXQoK018422 6, 17 -- for {Microscopy-at-Microscopy.Com} ; Thu, 9 Mar 2006 04:33:26 -0600 6, 17 -- Received: from vv-238.vv.slu.se (vv-238.vv.slu.se [194.47.228.116]) 6, 17 -- by alnus.slu.se (8.12.11/8.12.11) with ESMTP id k29AXPYd021333 6, 17 -- for {Microscopy-at-Microscopy.Com} ; Thu, 9 Mar 2006 11:33:25 +0100 (CET) 6, 17 -- Message-Id: {6.2.5.6.0.20060309112709.01cbef08-at-vv.slu.se} 6, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 6, 17 -- Date: Thu, 09 Mar 2006 11:33:24 +0100 6, 17 -- To: Microscopy-at-Microscopy.Com 6, 17 -- From: Kerstin Brismar {Kerstin.Brismar-at-vv.slu.se} 6, 17 -- Subject: SEM - Cryo system for sale 6, 17 -- Mime-Version: 1.0 6, 17 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 6, 17 -- Content-Transfer-Encoding: 8bit 6, 17 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k29AXQoK018422 ==============================End of - Headers==============================
5 nm gold particles conjugated to antibodies are visible in "routine" TEM. Section staining doesn't necessarily have to be reduced to see the beads, although that can help. This isn't to say this is easy, but it can be done. A STEM or EELS would make the beads more identifiable, and zero-loss imaging in a TEM with EELs would mean very lightly stained, or unstained, OsO4 postfixed, sections could be examined. 3 nm might maybe just be doable, but I haven't tried. This is without any enhancement, Ag or otherwise. Highest spatial resolution is obtained if the gold particles are conjugated to primary antibodies.
Phil
} The ongoing discussion about contrast brings to mind another } question. If one wants to add enough heavy metal to label a singular } structure on a biological tissue thin section, how much metal is } required to obtain a useful signal on a standard TEM? Would a STEM } system allow one to "see" the structure with a lower amount of heavy } metal label? Or does an energy filtered electron microscope (like } the Zeiss 902) permit one to resolve smaller clusters? } } I remember that some gold-linked antibody probes used fairly small } gold clusters (11 atoms perhaps?) to improve penetration into the } section, but that these ABs were only made visible after silver } enhancement for routine TEM. When does a cluster of metal atoms } become resolvable in a minimally stained thin section? } -- } David H. Hall, Ph.D. } Center for C. elegans Anatomy } Department of Neuroscience } Albert Einstein College of Medicine } 1410 Pelham Parkway } Bronx, NY 10461 } } www.wormatlas.org } www.aecom.yu.edu/wormem } } phone 718 430-2195 } fax 718 430-2514 -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 24 -- From oshel1pe-at-cmich.edu Thu Mar 9 07:10:42 2006 4, 24 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29DAflY004701 4, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 07:10:41 -0600 4, 24 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 24 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k29Dn74l006366; 4, 24 -- Thu, 9 Mar 2006 08:49:07 -0500 4, 24 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 24 -- Thu, 9 Mar 2006 08:11:01 -0500 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {f06230900c035d8b5efa5-at-[141.209.160.132]} 4, 24 -- In-Reply-To: {200603082208.k28M82xx028864-at-ns.microscopy.com} 4, 24 -- References: {200603082208.k28M82xx028864-at-ns.microscopy.com} 4, 24 -- Date: Thu, 9 Mar 2006 08:10:37 -0500 4, 24 -- To: hall-at-aecom.yu.edu 4, 24 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 24 -- Subject: Re: [Microscopy] contrast and imaging small clusters of heavy 4, 24 -- metals 4, 24 -- Cc: Microscopy-at-microscopy.com 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-OriginalArrivalTime: 09 Mar 2006 13:11:01.0839 (UTC) FILETIME=[EA0C65F0:01C6437A] 4, 24 -- X-CanItPRO-Stream: default 4, 24 -- X-Spam-Score: -3.5 () L_EXCH_MF,PORN_RP_PENETRATIONS_ 4, 24 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Not strictly true... It depends on your examination goals. Here are two extreme, but not unusual, examples:
If I am looking for Pb somewhat deep below the surface (especially if the matrix contains S / Mo that can interfere with the low energy Pb lines), higher kV is indicated (20-30 kV). In this example I need the high kV to penetrate deeply and to excite the higher energy Pb lines. The higher energy Pb lines will better escape from the sample as well.
OTOH, if I am trying to identify micron size B4C crystals residing on a surface, low kV is indicated (2-5 kV). In this case, I desire low penetration to minimize excitation of the substrate and minimize dilution the response.
Regards, Woody White BWXT Services
==============================Original Headers============================== 6, 26 -- From nwwhite-at-bwxt.com Thu Mar 9 07:18:11 2006 6, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 6, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29DIA4O006130 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 07:18:11 -0600 6, 26 -- Received: from ([131.184.13.224]) 6, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.191130; 6, 26 -- Thu, 09 Mar 2006 08:17:51 -0500 6, 26 -- Received: from bwxslynpo01.BWXS.BWXTECH.NET ([131.184.13.4]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 6, 26 -- Thu, 9 Mar 2006 08:17:51 -0500 6, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 26 -- Content-class: urn:content-classes:message 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- charset="iso-8859-1" 6, 26 -- Subject: Higher vooltages for EDX 6, 26 -- Date: Thu, 9 Mar 2006 08:17:51 -0500 6, 26 -- Message-ID: {70E4EA352EC987489773D55B0FF83E1C1DF9B6-at-bwxslynpo01.BWXS.BWXTECH.NET} 6, 26 -- X-MS-Has-Attach: 6, 26 -- X-MS-TNEF-Correlator: 6, 26 -- Thread-Topic: Higher vooltages for EDX 6, 26 -- Thread-Index: AcZDe94JLa3zFWS0TMaYtFslIpVqIg== 6, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 6, 26 -- To: {Microscopy-at-microscopy.com} 6, 26 -- X-OriginalArrivalTime: 09 Mar 2006 13:17:51.0488 (UTC) FILETIME=[DE37E000:01C6437B] 6, 26 -- Content-Transfer-Encoding: 8bit 6, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k29DIA4O006130 ==============================End of - Headers==============================
I too had noticed that many assumption this was a TEM question. Few addressed SEM although voltage is certainly an issue there.
I would also remind the other posters that this was submitted via Ask-a-Microscopist. There is an advisory on the initial post to remind us to copy the original inquirer on the discussion since they probably do not subscribe to the list. Some of you have been copying Naomi, but a lot haven't. She is missing out on your advice. It is easy to forget that as the discussion winds on.
Of course, we hope she will be happy enough with the responses that she becomes a regular list member.
Warren Straszheim Materials Analysis and Research Laboratory Iowa State University ------------------------------------------------ X-from: jquinn-at-www.matscieng.sunysb.edu [mailto:jquinn-at-www.matscieng.sunysb.edu] Sent: Wed 3/8/2006 1:35 PM To: wesaia-at-iastate.edu
Hi,
I understand. It sounds like against the common sense.
But the intensity of the EDX signal only depends on the element itself and the probability of scattering events. We use a factor " cross section" to quantified such probability. Look at its expression in any TEM book you will see the higher the voltage, the smaller the cross section.
Or I like to consider this question physically in the following way: Electrons can be consider as many single waves. The higher their voltage, the shorter their wavelength and the smaller the "size" of every of them. Apparently the small ball can travel longer in certain specimen. Just like a car is much easier to get blocked by the traffic than a motorcycle. Of course when there are no policemen. :-)
Hopefully this may help.
Hongqi
At 03:52 AM 3/9/2006, you wrote: } Sorry if I digress a bit, but I am new to the field of } EDX. I thought that higher voltages gave a higher } signal in EDX, and so a higher sensitivity. } Is it not true? } } Stéphane } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } Hi, } } } } In SEM, you will lose surface sensitivity. } } } } In TEM, scattering cross sections decrease which is } } not good for EDX and EELS. } } } } Hongqi } } } } Dept. of Materials Science and Engineering } } Pennsylvania State University } } University Park, PA 16802 } } email: hud105-at-psu.edu } } } } } } ==============================Original } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com
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Please don't make the mistake of simply correlating X-ray emission with a single parameter,like accelerating voltage, in either SEM or TEM applications.
Your measured x-ray intensity, as a function of accelerating voltage, is a product of a number of factors which include the ionization cross-section, electron beam current, electron energy loss, the scattering pathlength, and absorption path length.
As the accelerating voltage changes all of these parameters will vary and you need to include all of them in any assessment x-ray intensity for a given set of experimental conditions. The quantity that decreases with accelerating voltage is #Ionizations/nA/unitpathlength. Even though this quantity decreases with accelerating voltage for a constant probe size, it is likely that you will measure a higher x-ray signal as the accelerating voltage increases.
For example, below the critical excitation energy (Ec) for a given shell the x-ray emission for an element will be zero. It then increases rapidly to a broad maximum somewhere at ~ 2-4x Ec. Once you exceed ~ 4*Ec there is indeed a decrease in the cross-section. However this decrease is NOT linear, nor is it inversely proportional to accelerating voltage. Instead it is inversely proportional to the relativistically corrected energy of the electrons (1/2 mv^2), this means the decrease is not as great as you would expect. In addition, a number of electron sources actually yield higher beam currents at higher accelerating voltages, so even though the cross-section will be decreasing somewhat with accelerating voltage the net effect can be an increased x-ray signal, until such time as the depth of production is so great that the x-ray are absorbed within the sample before being detected.
If your bored and interested in seeing more detail on this for the TEM area, as well as some of the corresponding background and equations, go to the following URL
http://tpm.amc.anl.gov/Lectures/
then download the PDF file
XEDSAEMShortCourse.pdf
and look at pages 30-33 & 44-65.
Of course there are other deliterious effects of higher accelerating voltage, but that is a different discussion entirely.
Nestor Your Friendly Neighborhood SysOp
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
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I understand your explanation, but the intensity of the signal (Y axis) in EDX does not depend on the nature of the material (this is the X axis), but on the number of times the same signal is read. This means that the intensity of the signal read by EDX depends on the number of electrons which hit a certain point on the sample, per unit of time. And this depends on the current. And least that's what I thought! :-D
Stéphane
--- Hongqi Deng {hud105-at-psu.edu} wrote:
} Hi, } } I understand. It sounds like against the common } sense. } } But the intensity of the EDX signal only depends on } the element itself and } the probability of scattering events. We use a } factor " cross section" to } quantified such probability. Look at its expression } in any TEM book you } will see the higher the voltage, the smaller the } cross section. } } Or I like to consider this question physically in } the following way: } Electrons can be consider as many single waves. The } higher their voltage, } the shorter their wavelength and the smaller the } "size" of every of them. } Apparently the small ball can travel longer in } certain specimen. Just like } a car is much easier to get blocked by the traffic } than a motorcycle. Of } course when there are no policemen. :-) } } Hopefully this may help. } } Hongqi } } } At 03:52 AM 3/9/2006, you wrote: } } Sorry if I digress a bit, but I am new to the field } of } } EDX. I thought that higher voltages gave a higher } } signal in EDX, and so a higher sensitivity. } } Is it not true? } } } } Stéphane } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } ---------------------------------------------------------------------------- } } } } } } Hi, } } } } } } In SEM, you will lose surface sensitivity. } } } } } } In TEM, scattering cross sections decrease which } is } } } not good for EDX and EELS. } } } } } } Hongqi } } } } } } Dept. of Materials Science and Engineering } } } Pennsylvania State University } } } University Park, PA 16802 } } } email: hud105-at-psu.edu } } } } } } } } } ==============================Original } } } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam } protection around } } http://mail.yahoo.com } }
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==============================Original Headers============================== 8, 20 -- From nizets2-at-yahoo.com Thu Mar 9 11:24:18 2006 8, 20 -- Received: from web37407.mail.mud.yahoo.com (web37407.mail.mud.yahoo.com [209.191.87.60]) 8, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k29HOHJk015444 8, 20 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 11:24:17 -0600 8, 20 -- Received: (qmail 54719 invoked by uid 60001); 9 Mar 2006 16:52:12 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=3kV/Nj+ncCCzrH6eYWFGstUn2UqHFAmOL9udYdlBDwVnGDQeQu0C+uCgHk+DzZS9fIs/tD4mn2DLSy/94rPK7qwUq4XX4ed7Wn9d3pwQj6OaHFg9fyGw53GF194WbW+FjG46qDCbVLZhe5sv4nedawZwRsQfrDNqWRNfn5EEAa0= ; 8, 20 -- Message-ID: {20060309165212.54717.qmail-at-web37407.mail.mud.yahoo.com} 8, 20 -- Received: from [80.122.101.102] by web37407.mail.mud.yahoo.com via HTTP; Thu, 09 Mar 2006 08:52:12 PST 8, 20 -- Date: Thu, 9 Mar 2006 08:52:12 -0800 (PST) 8, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 20 -- Subject: Re: higher voltages for EDX 8, 20 -- To: Hongqi Deng {hud105-at-psu.edu} 8, 20 -- Cc: microscopy-at-microscopy.com 8, 20 -- In-Reply-To: {6.0.0.22.2.20060309102326.019b98f8-at-email.psu.edu} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
You are partially correct. You are confusing total number of counts (which is what you are talking about) with the physics of x-ray generation, which is dependant upon both material and experimental conditions.
Counting longer only improves the statistics it will not increase the number of x-rays per electron per unit pathlength.
Nestor Your Friendly Neighborhood SysOp
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
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==============================Original Headers============================== 11, 16 -- From zaluzec-at-aaem.amc.anl.gov Thu Mar 9 11:33:20 2006 11, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 11, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29HXJC9018026 11, 16 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 11:33:19 -0600 11, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 11, 16 -- by aaem.amc.anl.gov (8.12.11/8.12.10) with ESMTP id k29HXJ61001218 11, 16 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 11:33:19 -0600 11, 16 -- Mime-Version: 1.0 11, 16 -- Message-Id: {p06110405c03616f433bc-at-[146.139.72.105]} 11, 16 -- Date: Thu, 9 Mar 2006 11:33:18 -0600 11, 16 -- To: microscopy-at-microscopy.com 11, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 11, 16 -- Subject: Re: higher voltages for EDX 11, 16 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 11, 16 -- Content-Transfer-Encoding: 8bit 11, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k29HXJC9018026 ==============================End of - Headers==============================
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Email: pedromfjcosta-at-gmail.com Name: Pedro Costa
Organization: University of Cambridge
Title-Subject: [Filtered] EDX
Question: I have been using the ES-Vision software, from EMISPEC, to do some quantitative analysis of STEM-EDX spectra taken with a 200 kV microscope. Besides allowing the input of experimental parameters (energy resolution, sample thickness / orientation, etc), this software is capable to do background subtraction, peak fitting, modelling of spectra and extract quantitative details for each element. As regards the quantification procedure, the final table of results shows several parameters such as weight percentage composition, atomic percentage composition and an uncertainty percentage. In case someone has used this software for the same purposes, would it be possible to clarify how does ES-Vision work out the uncertainties in the calculations?Also, what are the formulas used for the elemental percentages calculations (weight and atomic)?
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Email: mbisher-at-princeton.edu Name: Margaret Bisher
Organization: Princeton University
Title-Subject: [Filtered] Light Microscopy of Wood
Question: Hello Group,
I have a graduate student here who wants to image wood. It seems simple, but to her (and me, being a TEM person) it's not. She only wants to do light microscopy, so thick sections only. We can do paraffin or plastic embedding, or even cryo (we have a cryostat as well as a microtome).
My question to the group, is what do we do? We tried a standard dehydration with ethanol and zylene into paraffin, but the paraffin did not seem to penetrate completely into the wood and when cutting, the wood seems to crumble and just not be what we are hoping for.
Any suggestions would be really great. I've never done plant material, so this is really foreign to me.
The sticks are really tiny, most of them are going to be anywhere from 2 to 5 mm in diameter, not huge stalks..... she ultimately would like cross sections of these.
Margaret E. Bisher Electron Microscopy Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113B Princeton, NJ 08544-1014 Office: (609) 258-7026 Fax: (609) 258-8468 email: mbisher-at-molbio.princeton.edu
Get a book on botanical microtechnique before you do anything, it will save you a lot of wasted time. The 'woodies' I knew all used celloidion (parlodion) embedding.
Geoff
mbisher-at-princeton.edu wrote:
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-- -- ********************************************** 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 **********************************************
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What you do get at higher kV is a better peak-to-background ratio (at least in a TEM).
Characteristic X-rays are emitted isotropically. However, part of the background arises from bremstralung which is forward scattered (i.e. down the column) - the degree of forward scattering is dependent on the velocity of the electrons. Hence higher kVs result in the bremstralung forward scattering increasing. But, since the EDX background is not wholly dependent on bremstralung, the actual instrumental gain is not as much as you would expect from a simple physics argument.
In the case of SEM, you are probably best going to low kV, since this reduces the excitation volume, so improving the spatial resolution. However, this only really works with a FEG gun (to get enough probe current at low kV) and with WDX, since you have to work with L and M lines and need the resolution of WDX to separate the lines.
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Email: martimor-at-nmsu.edu Name: M. M.
Title-Subject: [Filtered] RE: propylene oxide
Question: I was wondering if anyone could answer a question that I have been wondering about? I was told by a technician by a certain company that acetone can act as a "scavenger" (okay?) when used as a transitional solvent for Araldite 502/Embed-812 medium and propylene oxide would always be better. Why would this be? I am seeking any advice on this matter please.
Does anyone in Australia have a video display unit suitable to fit a Hitachi H-600 TEM that they're willing to part with? Our second unit is having a near-death experience.
Thanks,
John Brealey Queen Elizabeth Hospital EM Unit IMVS - TQEH Pathology Adelaide, South Australia (08) 8222 6612
john.brealey-at-imvs.sa.gov.au
==============================Original Headers============================== 6, 35 -- From john.brealey-at-imvs.sa.gov.au Thu Mar 9 17:01:39 2006 6, 35 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 6, 35 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29N1bnY032162 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 9 Mar 2006 17:01:38 -0600 6, 35 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 6, 35 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k29N1ZZr004923 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:35 +1030 (CST)' 6, 35 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 6, 35 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k29N1ZgP004906 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:35 +1030 (CST)' 6, 35 -- Received: from localhost (mesh.imvs.sa.gov.au [10.20.98.37]) 6, 35 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id EABF72EC03E 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:34 +1030 (CST) 6, 35 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 6, 35 -- by localhost (mesh.imvs.sa.gov.au [10.20.98.37]) (amavisd-new, port 10024) 6, 35 -- with LMTP id 31177-01-20 for {Microscopy-at-MSA.Microscopy.Com} ; 6, 35 -- Fri, 10 Mar 2006 09:31:34 +1030 (CST) 6, 35 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 6, 35 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id B81C92EC024 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:34 +1030 (CST) 6, 35 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 6, 35 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 6, 35 -- Subject: TEM Hitachi H-600 6, 35 -- Date: Fri, 10 Mar 2006 09:31:34 +1030 6, 35 -- Message-ID: {000201c643cd$6979e270$c88a140a-at-iqe36042} 6, 35 -- MIME-Version: 1.0 6, 35 -- Content-Type: text/plain; 6, 35 -- charset="iso-8859-1" 6, 35 -- Content-Transfer-Encoding: 7bit 6, 35 -- X-Priority: 3 (Normal) 6, 35 -- X-MSMail-Priority: Normal 6, 35 -- X-Mailer: Microsoft Outlook 8.5, Build 4.71.2173.0 6, 35 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 6, 35 -- Importance: Normal 6, 35 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au ==============================End of - Headers==============================
I learned EM from John Luft around the time he pioneered use of Epon and propylene oxide. Four years later I diverted to acetone after being surprised and impressed by the results of ROBERTSON, J. D., BODENHEIMER, T. S., and STAGE, D. E., J. Cell Biol., 1963, 19, 159. Still later i tested 60 degree C heat-cure of 10 ml test-tubes filled with my Araldite 506 embedding mixture deliberately adulterated by inclusion of 15-20% solvent, trying propylene oxide, acetone, EtOH and MeOH. I found no reason then or since to give up acetone in favor of the others. Specifically, the propylene oxide mix cured to give a softer and somewhat cheesy polymer; the acetone mix reduced somewhat in volumed during cure and the cured product was similar to unadulterated resin. I can't recall the alcohols results; I think they were similar to acetone? . I was surprised that the propylene oxide result was inferior to the acetone result; I had expected both to evaporate during cure to leave a final polymer unaltered y the solvent inclusion.
I never heard the term "scavenger" applied. Luft called propylene oxide a "reactive diluent", and suggested that any small amount that failed to evaporate during heat-cure would incorporate harmlessly in the polymer. He was probably correct. My point is that large amounts of propylene oxide are not so "harmless", while similarly large amounts of included acetone are surprisingly innocuous. -mike reedy-
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-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
Hello, I was wondering if anyone had used the Navitar Video Fluorescence Scope: http://www.navitar.com/zoom/zfl_gen.htm
I was considering using it with some bacteria fluorescence probes and was wondering if anyone had tried it out and what they thought of it. Any advice appreciated.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 3, 24 -- From gvrdolja-at-nature.berkeley.edu Fri Mar 10 19:15:31 2006 3, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 3, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2B1FVxH012780 3, 24 -- for {microscopy-at-microscopy.com} ; Fri, 10 Mar 2006 19:15:31 -0600 3, 24 -- Received: from localhost (localhost [127.0.0.1]) 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id AFBD3C1E4A 3, 24 -- for {microscopy-at-microscopy.com} ; Fri, 10 Mar 2006 17:15:30 -0800 (PST) 3, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 3, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 3, 24 -- with ESMTP id 17190-09 for {microscopy-at-microscopy.com} ; 3, 24 -- Fri, 10 Mar 2006 17:15:30 -0800 (PST) 3, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 3, 24 -- id 42264C1E4C; Fri, 10 Mar 2006 17:15:30 -0800 (PST) 3, 24 -- Received: from localhost (localhost [127.0.0.1]) 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 3A083C1E4A 3, 24 -- for {microscopy-at-microscopy.com} ; Fri, 10 Mar 2006 17:15:30 -0800 (PST) 3, 24 -- Date: Fri, 10 Mar 2006 17:15:29 -0800 (PST) 3, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 3, 24 -- To: microscopy-at-microscopy.com 3, 24 -- Subject: question about Navitar fluorescent microscope 3, 24 -- Message-ID: {Pine.SOC.4.64.0603101713260.13399-at-nature.Berkeley.EDU} 3, 24 -- MIME-Version: 1.0 3, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 3, 24 -- X-Virus-Scanned: amavisd-new at nature.berkeley.edu ==============================End of - Headers==============================
I notice that Epson has just started shipping the V700 [and V750 Pro] large format flatbed scanner that costs around £400 to £550 [for the Pro version that has enhanced optics and Silverfast Ai] in the UK. The V700 is a top end consumer model and the V750 is a 'professional' model (and so offers more).
Check it out at: http://www.photo-i.co.uk/News/Feb06/Epson_V700_scanner.htm and click the 'interactive review'.
This scanner is clearly a real advance on the slightly cheaper Epson 4990F and Canon 9950F scanners that have 4,800 dpi. The V700 [and V750] is rated at 6,400 dpi. The new Epson V750 Pro produces sharper focus in its scans than these previous models that gave quite 'soft' images - this is no doubt due to better optics.
X-from the review link above it is clear that the V700 series scanner is a significant improvement on the last generation of prosumer flat bed scanners and should be a serious contender for any shortlist on those wishing to scan large format negatives/positives up to A4 in size (i.e. TEM negatives).
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
Is this site truely independent or is it the voice of the manufacturer or industry group? Has anyone taken a TEM negative and scanned it on the new v. the old scanners? And does anyone need a TEM negative scanned at 6400 dpi? I would rather see an real increase in the dynamic range rather than a dpi "race".
Geoff
keith.morris-at-ucl.ac.uk wrote:
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==============================Original Headers============================== 6, 32 -- From mcauliff-at-umdnj.edu Mon Mar 13 09:08:18 2006 6, 32 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 6, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2DF8IMg010097 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Mar 2006 09:08:18 -0600 6, 32 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 6, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id B4ECD4BE3D 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Mar 2006 09:08:17 -0600 (CST) 6, 32 -- Received: from polaris.umdnj.edu (polarisb.UMDNJ.EDU [130.219.34.133]) 6, 32 -- by zix02.umdnj.edu (Proprietary) with ESMTP id 869DD4BE31 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Mar 2006 09:08:16 -0600 (CST) 6, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 32 -- id {0IW200I01N6CAI-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 32 -- for microscopy-at-msa.microscopy.com; Mon, 13 Mar 2006 10:08:15 -0500 (EST) 6, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 6, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 32 -- 2004)) with ESMTP id {0IW200GSINCH7J-at-Polaris.umdnj.edu} ; Mon, 6, 32 -- 13 Mar 2006 10:07:31 -0500 (EST) 6, 32 -- Date: Mon, 13 Mar 2006 10:06:37 -0500 6, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 32 -- Subject: Re: [Microscopy] Scanner for TEM Micrographs 6, 32 -- In-reply-to: {200603131321.k2DDLqdn024056-at-ns.microscopy.com} 6, 32 -- To: keith.morris-at-ucl.ac.uk, 6, 32 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 32 -- Message-id: {44158A7D.3000902-at-umdnj.edu} 6, 32 -- MIME-version: 1.0 6, 32 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 32 -- Content-transfer-encoding: 8BIT 6, 32 -- X-Accept-Language: en-us, en 6, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 32 -- Gecko/20040804 Netscape/7.2 (ax) 6, 32 -- References: {200603131321.k2DDLqdn024056-at-ns.microscopy.com} ==============================End of - Headers==============================
Dear All John Benjamin Dancer produced photomicrographs of fleas in the mid-19th century. Was he the very first person to record a microscope image using photography?
Who was the first person to make a photomicrograph of a protein crystal?
Best wishes Chris
Dr Christopher E. Jeffree University of Edinburgh Institute of Molecular Plant Sciences King's Buildings, Mayfield Road Edinburgh, EH9 3JH Scotland, UK Tel: +44 131 650 5554 FAX: +44 131 650 5392 email c.jeffree-at-ed.ac.uk
==============================Original Headers============================== 5, 26 -- From c.jeffree-at-ed.ac.uk Mon Mar 13 10:04:44 2006 5, 26 -- Received: from lawnmarket.ucs.ed.ac.uk (lawnmarket.ucs.ed.ac.uk [129.215.166.63]) 5, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2DG4h0w024583 5, 26 -- for {microscopy-at-microscopy.com} ; Mon, 13 Mar 2006 10:04:43 -0600 5, 26 -- Received: from EMFBIO (emf.icmb.ed.ac.uk [129.215.156.56]) 5, 26 -- by lawnmarket.ucs.ed.ac.uk (8.12.10/8.12.10) with SMTP id k2DG4gNH022706 5, 26 -- for {microscopy-at-microscopy.com} ; Mon, 13 Mar 2006 16:04:42 GMT 5, 26 -- Message-ID: {000e01c646b7$51009530$389cd781-at-EMFBIO} 5, 26 -- Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} 5, 26 -- From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} 5, 26 -- To: {microscopy-at-microscopy.com} 5, 26 -- Subject: First photomicrograph 5, 26 -- Date: Mon, 13 Mar 2006 16:00:57 -0000 5, 26 -- MIME-Version: 1.0 5, 26 -- Content-Type: text/plain; 5, 26 -- format=flowed; 5, 26 -- charset="iso-8859-1"; 5, 26 -- reply-type=original 5, 26 -- Content-Transfer-Encoding: 7bit 5, 26 -- X-Priority: 3 5, 26 -- X-MSMail-Priority: Normal 5, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 5, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 5, 26 -- X-Edinburgh-Scanned: at lawnmarket.ucs.ed.ac.uk 5, 26 -- with MIMEDefang 2.33, Sophie, Sophos Anti-Virus 5, 26 -- X-Scanned-By: MIMEDefang 2.33 (www . roaringpenguin . com / mimedefang) ==============================End of - Headers==============================
To be honest most of us here use only 800 to 1,200 dpi for TEM negative scanning anyway, whether for publication or image analysis (at this dpi the Epson 9950F will no doubt produce similar scans to the new Epson V750). We may zoom in on areas for scanning (enlargements), but never 'archive' the whole TEM negative as digitised images - we just keep the negatives. Our new Epson 9950F flatbed scanner we use (cost £280 in 2006) produces digitised images scanned at maximum resolution that are distinguishable to those from our Agfa DuoScan flatbed scanner (that cost £5,000 in 2001). Both scanners were at their maximum usable resolution of 2,400 dpi and 2,500 dpi respectively. I have noticed that at 4800 dpi the Epson 4990 Photo has juddered once while scanning - this might be due to inferior mechanics or poor vibration protection, the heavy DuoScan has a 'squash ball' type isolation feature. The DuoScan is slower and noisier though, and can only scan one negative at a time in it's 'sweet spot', compare to three to six negatives with the Epson 4990 Photo (it's also SCSI rather than universal USB2). Image morphometry distortion is also very low across the platter with the Epson, producing mean errors in length and area measurements of around 0.3% at 800 dpi and 0.15% at 2,400 dpi (using graph paper as a target). This is very close to the inherent errors from using the mouse in MetaMorph.
Images from both scanners need a quick bit of Photoshop work to get them looking their best. We used to spend hours doing that when printing EM photographs in the dark room - now with these cheap multi-purpose flatbed scanners you should never have to do that again.TEM negatives go up to a maximum of around six times enlargement, although at high TEM magnifications the resin grain may be more of a problem than the film grain. I'm also scanning a few B&W TEM negatives and colour 35mm slides on UCL's 8000 dpi Imacon professional scanner at UCL's Reprographics for comparison - that's £5 to £10 (} 50Mb) per scan though depending on image file size.
DMax in these modern cheap flatbed scanners is reported to be around 3.8 to 4.0 (its difficult to compare specs though as, like with dpi, manufacturers lie about the true value differently). Correctly exposed B&W silver halide negatives are reported to have a DMax of nearer 1.5 compared to a colour dye slides 3.5D - so a good scanners DMax is less of a problem with B&W negatives and even a dog of a modern scanner should cope with most TEM negatives in terms of dynamic range. Plus we can only distinguish 191 grey levels, so 8-bit (256 greys) rather than 14-bit (16,385 greys) is mostly fine for B&W photographs - we do a bit better with colour and so need things like CMYK printing and ICC profiling for this. As with microscopes, fantastic resolution isn't much use if there's no contrast, but again as with microscopes, increased contrast often has the cost of reducing resolution. B&W TEM negatives that initially look good with very high contrast (DMax nearer 2.4) are generally inferior in detail to negatives that have a far more neutral tonal balance. One problem with TEM negatives is that we can't immediately tell if the picture is poor after zooming in, particularly if a cheap scanner secretly applies USM, whereas if its a colour scan of our kids faces, or writing on the side of a ship, it's immediately obvious. For TEM negatives it's easier just to compare the results from different scanners and with the manual view looking at the negative with a light box and a magnifier. Although we don't need a [probably optimistic] '6,000 dpi' for large format negatives unless we really want to look at the film grain, it does suggest that the scanner has very good optics for great lower resolution scans. The use of Digital ICE [FARE] dust removal is pretty irrelevant for B&W negatives as the process is optimised for colour film only.
It naturally tends to be photographers who want the higher resolutions of 4,000 dpi and above, plus sharp focussing, largely for the archiving smaller 35mm colour slide or negatives. Here resolving detail in shadow with low noise [i.e. high DMax] is very important - further helped by Photoshop CS's great 'shadow/highlight' feature. My colour slides of the family from the 1950's to 1980's are all showing signs of aging, in particular the 50's slides that have now gone very dark brown (although a few Fuji slides from the 70's have also really aged badly - fortunately I mainly used Afga/Perutz back then). I now wish I hadn't got myself the Canon 9950F (£260) for home use at Christmas - the new Epson V750 would have made a better job of digitising my family colour slides and photos (although I probably wouldn't notice the difference much at A4 after USM, and the V750 is £200 more). At work we are happy to continue with the Epson 4990 Photo, although I obviously would have bought the Epson V750 with its sharper optics if I was buying now - the Epson 4990 Photo and Canon 9950F do definitely produce 'softer' slightly out of focus scans (and their output quality is identical). Also don't buy the Canon 9950F to scan TEM negatives, it can't do 'A4' negative scans and is restricted to the sizes of the film holders (that are 4 x 5" and 120 and not standard EM negative sizes). The Epson 4990 (and the near identical V750) can scan negatives to nearly the full platter size - that means three or six TEM negatives in one go (depending on their size).The V750 can also scan more 35mm slides and negatives in one go than the 4990. We have plenty of 35mm film to scan here from old optical photo-microscopes, 'talks' and lab cameras. My Canon 9950F flatbed scanner at home produces better colour scan images, particularly from colour negatives, than my comparably priced dedicated Acer [Benq] ScanWit 2740s 2,700 dpi 35mm slide scanner - no doubt the Epson V750 flatbed would do even better.
I suppose we should use 'acid free' bag storage to protect our colour negatives, photographs and slides from atmospheric pollution and decay - just the same as the valuable linen, comic and book crowd do - but I've never got around to it. Fortunately time has demonstrated that the silver halide process produces a far more stable image, albeit black and white, compared to those produced with colour dyes. The support material though, e.g particularly old cellulose nitrate stock, may degrade badly with time. Early photographers fortunately used glass plate as the support medium that was very durable [until you drop them]. I have a few 1920's large format B&W film negatives of the family and 1930's16mm B&W movie film (Pathe News) that still look good though.
Keith
PS. The http://www.photo-i.co.uk site is an independent one run in the UK. The site is quite similar to the excellent http://www.dpreview.com for digital cameras. It's quite clear from their reviews that they are independent - although they are naturally keen photographers rather than electron microscope users so their priorities may differ. If they say some aspect of the product will really pig you off - it invariably does. I expect they do consultancy reviews for magazines and that manufacturers are keen to get them to review their product if they think it's good. It's also rather obvious that in short review articles in the like those of PCPro magazine [in the UK] the reviewers have often spent hardly any time trying to get the best out of the scanner.
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
My last posting missed out a rather key 'in' from 'in'distguishable (I should have stuck with the less technically correct term 'identical', as in the my first draft). The sentence in paragraph one, line three, thus should have read:
"Our new Epson 9950F flatbed scanner we use (cost £280 in 2006) produces digitised images scanned at maximum resolution that are indistinguishable to those from our Agfa DuoScan flatbed scanner (that cost £5,000 in 2001)."
i.e. both scanned images look pretty much the same and you can't tell them apart once you have put them through Photoshop's autocontrast adjustment. Prior to Photoshop editing, the 'auto' setting on the older Agfa DuoScan twain interface produced quite light scans that actually looked worse than the cheap Epson 9950F's more contrasty ones.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
I have always made my Epon with...well Epon. Now I come in a lab (i am the only EM here) where we have stocks of Glycid ether. Are they similar products? Should I use it the same way as Epon and at the same proportions?
Stephane (without i ;-))
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Tue Mar 14 10:42:38 2006 7, 18 -- Received: from web37408.mail.mud.yahoo.com (web37408.mail.mud.yahoo.com [209.191.87.61]) 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2EGgc7H026412 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 14 Mar 2006 10:42:38 -0600 7, 18 -- Received: (qmail 36782 invoked by uid 60001); 14 Mar 2006 16:42:38 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=SGVnigD362HyNBBpWqulINdx4DZuhpYEH/KXo23eBg0s9tANhKguMtkdM5HYPG2SwQBMv0F2knf6QxcjiN3EvhoqAmnK0AnEpPFdsd97p+/a88DqJ0yy4Zd5qFq+V89n7jiQo3B9Mq/vZjLSa5mK51I+Hv06akJlSUS8nczo9Mw= ; 7, 18 -- Message-ID: {20060314164238.36780.qmail-at-web37408.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via HTTP; Tue, 14 Mar 2006 08:42:38 PST 7, 18 -- Date: Tue, 14 Mar 2006 08:42:38 -0800 (PST) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: glycid Ether 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; charset=iso-8859-1 7, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
} -----Original Message----- } From: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Tuesday, March 14, 2006 10:50 AM } To: Dusevich, Vladimir } Subject: [Microscopy] glycid Ether } } } } } -------------------------------------------------------------- } -------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } -------------- } } Dear all, } } I have always made my Epon with...well Epon. Now I come in a } lab (i am the only EM here) where we have stocks of Glycid ether. } Are they similar products? Should I use it the same way as } Epon and at the same proportions? } } Stephane (without i ;-)) } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection } around http://mail.yahoo.com } } ==============================Original } Headers============================== } 7, 18 -- From nizets2-at-yahoo.com Tue Mar 14 10:42:38 2006 7, } 18 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.87.61]) } 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP } id k2EGgc7H026412 } 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 14 Mar } 2006 10:42:38 -0600 } 7, 18 -- Received: (qmail 36782 invoked by uid 60001); 14 Mar } 2006 16:42:38 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; } q=dns; c=nofws; } 7, 18 -- s=s1024; d=yahoo.com; } 7, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Conten t-Type:Content-Transfer-Encoding; } 7, 18 -- } b=SGVnigD362HyNBBpWqulINdx4DZuhpYEH/KXo23eBg0s9tANhKguMtkdM5HY } PG2SwQBMv0F2knf6QxcjiN3EvhoqAmnK0AnEpPFdsd97p+/a88DqJ0yy4Zd5qF } q+V89n7jiQo3B9Mq/vZjLSa5mK51I+Hv06akJlSUS8nczo9Mw= ; } 7, 18 -- Message-ID: } {20060314164238.36780.qmail-at-web37408.mail.mud.yahoo.com} } 7, 18 -- Received: from [80.122.101.102] by } web37408.mail.mud.yahoo.com via HTTP; Tue, 14 Mar 2006 } 08:42:38 PST 7, 18 -- Date: Tue, 14 Mar 2006 08:42:38 -0800 } (PST) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 } -- Subject: glycid Ether 7, 18 -- To: } microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- } Content-Type: text/plain; charset=iso-8859-1 7, 18 -- } Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== }
==============================Original Headers============================== 6, 23 -- From DusevichV-at-umkc.edu Tue Mar 14 11:29:02 2006 6, 23 -- Received: from KC-MSXPROTO2.kc.umkc.edu (pop3.exchange.umkc.edu [134.193.143.155] (may be forged)) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2EHT0qX002997 6, 23 -- for {microscopy-at-msa.microscopy.com} ; Tue, 14 Mar 2006 11:29:01 -0600 6, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by KC-MSXPROTO2.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Tue, 14 Mar 2006 11:28:59 -0600 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: RE: [Microscopy] glycid Ether 6, 23 -- Date: Tue, 14 Mar 2006 11:28:58 -0600 6, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADBF9-at-KC-MSX1.kc.umkc.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [Microscopy] glycid Ether 6, 23 -- Thread-Index: AcZHh08url+foB08SoSaavJe2GWPwQABWlcA 6, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 6, 23 -- To: {microscopy-at-msa.microscopy.com} 6, 23 -- X-OriginalArrivalTime: 14 Mar 2006 17:28:59.0657 (UTC) FILETIME=[C79CBF90:01C6478C] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2EHT0qX002997 ==============================End of - Headers==============================
The University of California at Santa Barbara (Molecular, Cellular, and Developmental Biology Department and the Neuroscience Research Institute) is offering a workshop on Advanced Microscopy Digital Imaging. This course is co-sponsored with Purdue University and will be held in Santa Barbara from April 24-28th. In addition to our academic sponsors, the workshop has the generous support of Media Cybernetics who is providing their software and support for a teaching assistant. Also, Olympus of America is providing four inverted microscope stands, a DSU (Disk Scanning Unit) and Fluoview scanning confocal microscope. There will be digital cameras from Q-Imaging, fluorescent filters from Omega Optical, cell injectors from Eppendorf, and stage heaters and live cell chambers from Bioptechs. For more information about the course, please check the following web site:
I have an Olympus BH-2 Microscope that requires a Polariser/Analyser set and filters for both light paths. I have been advised by the local Olympus reps that the microscope has been superseded many years ago and the parts are hard to come by.
Does anyone have any stashed in a draw that they would be willing to part with or know of any third party suppliers.
Any help would be greatly appreciated.
Regards George
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==============================Original Headers============================== 10, 27 -- From George.Theodossiou-at-amcor.com.au Tue Mar 14 18:02:05 2006 10, 27 -- Received: from aiti251.amcor.com.au (smtp.amcor.com.au [202.14.180.248]) 10, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2F022RK026584 10, 27 -- for {microscopy-at-msa.microscopy.com} ; Tue, 14 Mar 2006 18:02:03 -0600 10, 27 -- Received: from aadcex0001.amcor.net (unverified) by aiti251.amcor.com.au 10, 27 -- (Content Technologies SMTPRS 4.3.19) with ESMTP id 10, 27 -- {T7709ff7fcaa0de98c8bd0-at-aiti251.amcor.com.au} for 10, 27 -- {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 11:07:27 +1100 10, 27 -- Received: from aadcex0004.amcor.net ([160.222.80.235]) by 10, 27 -- aadcex0001.amcor.net with Microsoft SMTPSVC (6.0.3790.211); Wed, 15 Mar 10, 27 -- 2006 11:02:01 +1100 10, 27 -- Received: from 160.222.214.38 ([160.222.214.38]) by aadcex0004.amcor.net 10, 27 -- ([160.222.80.235]) with Microsoft Exchange Server HTTP-DAV; Wed, 15 Mar 10, 27 -- 2006 00:02:00 +0000 10, 27 -- User-Agent: Microsoft-Entourage/11.2.1.051004 10, 27 -- Date: Wed, 15 Mar 2006 11:01:25 +1100 10, 27 -- Subject: Filters For Olympus BH-2 10, 27 -- From: "George.Theodossiou" {George.Theodossiou-at-amcor.com.au} 10, 27 -- To: {microscopy-at-msa.microscopy.com} 10, 27 -- Message-ID: {C03DA485.101C%George.Theodossiou-at-Amcor.com.au} 10, 27 -- Thread-Topic: Filters For Olympus BH-2 10, 27 -- Thread-Index: AcZHw5m62ByWbLO2EdqaGwANkzYUMg== 10, 27 -- Mime-version: 1.0 10, 27 -- Content-type: text/plain; charset="US-ASCII" 10, 27 -- Content-transfer-encoding: 7bit 10, 27 -- X-OriginalArrivalTime: 15 Mar 2006 00:02:01.0242 (UTC) 10, 27 -- FILETIME=[AF550FA0:01C647C3] ==============================End of - Headers==============================
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Email: maloneyb-at-fiu.edu Name: Barbara
Organization: FIU
Title-Subject: [Filtered] How to change filament in Phillips 300 TEM
Question: I have changed filaments in other instruments, but never in this older model. The manual doesn't seem to cover this - does anyone have the written procedure. Really would appreciate it greatly. Thanks Barbara
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Email: pekysar-at-ucdavis.edu Name: Pat Kysar
Organization: University of California, Davis
Title-Subject: [Filtered] Low Dose TEM
Question: Hello all, We have a client who is trying to use the low dose function of our FEI CM120. He is looking at magnetic nano particles on a substrate. He was able to make it work but is still getting damage to his sample. Unfortuately, neither of the techs here have used the low dose so we are quite unfamiliar with it and aren't much help. He would like to know what radius he should be using and we would like to hear from anyone who has experience with the low dose on this tool (or any tips, for that matter) which might help him achieve success. We would all appreciate any help! Cheers, Pat Kysar University of California, Davis Medical School, Pathology EM Lab
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Email: biology-at-ucla.edu Name: Eric
Organization: UCLA Medical Center
Title-Subject: [Filtered] Cleaning Grids
Question: This was never a problem till about a month ago..
Since about a month we have been having a problem keeping the sections stuck to the grids. The grids are cleaned in 100% Acetone and dried. Sections are picked up either from above or below the water.
When the grids are stained using the microwave staining method we have been using for several years the sections come off the grids... Every now and then the sections will stick to the grids and everyhting is fine.
Any suggestions about consistency? I have tried staining less time in the microwave, but this does not make a difference...
Any different ways to clean the grids so the sections will stick better?
Dear Stephane, ... the problem with HV and 'sensitivity' of EDX measurements is very complex, as Nestor already stated. First, the overvoltage U = Eo/Ec (primary electron energy / critical shell ionisation energy) should be at least more than 2 for all elements of interest. If not, every gain in HV is leading to extreme excitation enhancements of the element (and X-ray production), like Nestor already stated. If U } 3, the X-ray excitation curve decreases quite flat.
If the basic excitation of characteristic X-rays is sufficient:
#1 } From the point of view of detection limits you have to consider the peak to background ratio. The background in EPMA is bremsstrahlung. The ratio of characteristic line excitation to bremsstrahlung excitation is always gaining with primary electron energy (HV). Therefore you achieve always better detection limits with higher HV (but see #2, the count rate limitation can work against!)
#2 You must take into account: Higher electron energy do always excite much more higher energy X-rays (both characteristic X-rays and bremsstrahlung). The count rate possibilities of an EDX are always limited. So you have to consider, that you possibly will get less count rates for the elements of interest with higher energy excitation... because your pulse processor has to process more high energy X-ray signals (less counts in a given time for the low energy X-rays you have in focus). E.g. The peaks are lower in a Au/Ag alloy between 2..4 keV (Ag-L/Au-M) with 40 kV excitation, than they would be with 15 kV. But the Au-L lines (near 10 keV) are 4 times higher (because 15 keV is very low for Au-L). Therefore, an increase of HV is bad for Ag-L/Au-M measurement, if a limited EDX count rate is given.
#3 Also the X-ray absorption in specimen is increasing with higher HV. Particularly weaker element counts will be very much reduced behind absorption jump of a main element. But to drive against absorption: Tilt the specimen towards the EDX detector. The absorption influence is not so important with TEM X-ray measurements (thin specimen).
Finally: The better detection limits ('sensitivity') goes really with higher HV. But one has to take into account limitations in EDX pulse processing and absorption issues (not in general, depends of matrix elements in specimen). Therefore a simple specimen tilt is often much better than an increase of excitation energy to improve count rate yield of an element of interest. If the count rate from specimen is sufficient, the use of a shorter pulse processor shaping-time increases the detection sensitivity (despite the worse energy resolution), because you can detect more counts in same time.
Only spectra simulation software is able to answer these complex excitation and absorption influences in advance, without any data acquisition for tests.
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Sorry if I digress a bit, but I am new to the field of EDX. I thought that higher voltages gave a higher signal in EDX, and so a higher sensitivity. Is it not true?
Not a lot of people know the answer to this, apparently Any advance on one reply??
Best wishes Chris
----- Original Message ----- X-from: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: {microscopy-at-microscopy.com} Sent: Monday, March 13, 2006 4:00 PM
Obviously photo-microscopy was going strong in 1904. There's no direct link to the article, so to find a rather incredible picture of 10 feet of bellows and plate camera connected to a simple brass compound microscope goto :
http://www.microscopy-uk.org.uk/
Click main resources ' Micscape article library'. Then click 'find' and enter '1904'
You will then get the link : 'Taking a photomicrograph .... in 1904 by Dave Walker, UK'
I think I may even be able to see a protein crystal on the microscope stage.
Any takers for an earlier example ?
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {c.jeffree-at-ed.ac.uk} To: {keith.morris-at-ucl.ac.uk} Sent: Wednesday, March 15, 2006 9:24 AM
Well, it's kind of a trick question. Would a photomicrograph of a protein containing structure showing birafrigence count? Would a X-ray defraction photo taken with a "micro" camera count? Ahh.. these are the question to settle over a beer or a nice cup of coffee.....
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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c.jeffree-at-ed.ac.u k To: frank.karl-at-degussa.com cc: 03/15/2006 03:55 Subject: [Microscopy] Fw: First photomicrograph AM Please respond to c.jeffree
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Not a lot of people know the answer to this, apparently Any advance on one reply??
Best wishes Chris
----- Original Message ----- X-from: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: {microscopy-at-microscopy.com} Sent: Monday, March 13, 2006 4:00 PM
Barbara,
I have a set of manuals for the EM 300. Instructions for changing the filament are in the "Operating Instructions" volume -- there are several separate volumes -- Section D, pg 171 ff. Sounds like you don't have this volume. I can photocopy the pages and send them to you if you need. Or, if you or someone else using a Philips EM 300 needs the manuals, I can send them off -- we don't have one of these anymore. Note to the list: I also have a manual for an RCA EMU 4, if someone needs one. Might cost a pint or two at the next M&M meeting.
Phil
} This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both maloneyb-at-fiu.edu as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: maloneyb-at-fiu.edu } Name: Barbara } } Organization: FIU } } Title-Subject: [Filtered] How to change filament in Phillips 300 TEM } } Question: I have changed filaments in other instruments, but never } in this older model. The manual doesn't seem to cover this - does } anyone have the written procedure. Really would appreciate it } greatly. } Thanks } Barbara -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 24 -- From oshel1pe-at-cmich.edu Wed Mar 15 07:22:54 2006 4, 24 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FDMrQQ010632 4, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 07:22:53 -0600 4, 24 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 24 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k2FE154l025572; 4, 24 -- Wed, 15 Mar 2006 09:01:05 -0500 4, 24 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 24 -- Wed, 15 Mar 2006 08:22:39 -0500 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {f06230903c03dc4bd1d51-at-[141.209.160.132]} 4, 24 -- In-Reply-To: {200603150410.k2F4ATW6001828-at-ns.microscopy.com} 4, 24 -- References: {200603150410.k2F4ATW6001828-at-ns.microscopy.com} 4, 24 -- Date: Wed, 15 Mar 2006 08:22:48 -0500 4, 24 -- To: maloneyb-at-fiu.edu 4, 24 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 24 -- Subject: Re: [Microscopy] viaWWW: How to change filament in Phillips 300 4, 24 -- TEM 4, 24 -- Cc: Microscopy-at-microscopy.com 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-OriginalArrivalTime: 15 Mar 2006 13:22:39.0309 (UTC) FILETIME=[88405FD0:01C64833] 4, 24 -- X-CanItPRO-Stream: default 4, 24 -- X-Spam-Score: -4 () L_EXCH_MF 4, 24 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
It appears that John Dancer does have the honour of the first photomicrograph (of the flea), as suggested by Chris. However, ironically it is for inventing the procedure to create microphotographs on microscope slides (microfilm) that he is most remembered. I can't find any illustration of his 1840's 'gas powered microscope' on the web though, only pictures of him, his wife and many children.
A quick cut and paste biography of John Benjamin Dancer :
In 1840, when he was 28, John Dancer showed the world's first photomicrograph, of a Flea, in Liverpool, and he subsequently developed the microphotograph technique. In July 1840 he made a daguerreotype photograph of the flea, using a gas-illuminated microscope (this was a positive image on a metal support - the Daguerreotype was the first successful photographic process, the discovery being announced on 7 January 1839). In 1852 Dancer started making microscopic photographs - tiny photographs which could be viewed through a microscope. The microphotographs soon became popular and Dancer developed a large catalogue including photographs of the Royal family and Niagara Falls. Micro- photographs were then sold at one shilling (5p) each, or ten shillings and six pence (52.5p) for a dozen. He produced them commercially from about 1857. Although they sold poorly at first, within a few years they had become much sought after by science enthusiasts. He worked on various subjects, including landscapes, the Ten Commandments, and his most prestigious commission was for Queen Victoria, for whom he produced 5 miniature photographs of her family which were set in a signet ring - each picture being no more than 1/8th inch in diameter, and which were magnified in the ring by means of a jewel lens which he personally had cut. Dancer sold some 500 microphotograph slides, many of which were of well known paintings in art galleries. Particularly popular were slides of members of the Victorian Royal Family, of Emperor Napoleon, and of an 1858 20 banknote.
Although treated as a novelty until the 1920s, the microphotographic process eventually became 'microfilm'. Utilizing John Dancer's techniques, a French optician, Rene Dagron, was granted the first patent for microfilm in 1859 and began the first commercial microfilming enterprise. Dagron, during the Franco-Prussian War, demonstrated a practical use for microforms when carrier pigeons were used to transport microfilmed messages across German lines.
Otherwise Dancer's invention was not taken seriously, being variously described as "being of little or no practical use" and "childish and trivial". Yet, today, this invention is now used widely in banks, libraries and archives as a method of keeping important materials in an efficient, space-saving and economical way. He also invented the stereoscopic camera which he patented in 1853, contacts for electric alarms and a new form of illumination and photo-transparencies for use in lantern slide projection. Although not confirmed, it is widely believed Dancer created the first magic lantern photographic slide.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
"Fox Talbot was making images of botanical specimens with his solar microscope as early as February 1835, and is often credited as being the first photomicrographer. He especially loved photographing crystals but of what type is mostly unknown today."
Nancy
Nancy Smythe Department of Otolaryngology Head and Neck Surgery Medical University of South Carolina 843-792-8835 843-792-0368 Fax
==============================Original Headers============================== 6, 27 -- From smythen-at-musc.edu Wed Mar 15 08:16:37 2006 6, 27 -- Received: from caerbannog.musc.edu (caerbannog.musc.edu [128.23.203.43]) 6, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FEGaph024908 6, 27 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 08:16:36 -0600 6, 27 -- Received: from revere3.musc.edu (revere3.musc.edu [128.23.203.9]) 6, 27 -- by caerbannog.musc.edu (8.13.4/8.12.10) with ESMTP id k2FEEs7f008469 6, 27 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 09:14:54 -0500 6, 27 -- Received: from cl.musc.edu (cl.emr.musc.edu [128.23.151.3]) 6, 27 -- by revere3.musc.edu (8.12.9/8.12.9) with ESMTP id k2FEErAl017128 6, 27 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 09:14:53 -0500 (EST) 6, 27 -- Received: from CL-MTA by cl.musc.edu 6, 27 -- with Novell_GroupWise; Wed, 15 Mar 2006 09:14:31 -0500 6, 27 -- Message-Id: {s417daf7.004-at-cl.musc.edu} 6, 27 -- X-Mailer: Novell GroupWise Internet Agent 6.0.4 6, 27 -- Date: Wed, 15 Mar 2006 09:14:28 -0500 6, 27 -- From: "Nancy Smythe" {smythen-at-musc.edu} 6, 27 -- To: {microscopy-at-msa.microscopy.com} 6, 27 -- Subject: Re first photomicrograph 6, 27 -- Mime-Version: 1.0 6, 27 -- Content-Type: text/plain; charset=US-ASCII 6, 27 -- Content-Disposition: inline 6, 27 -- X-MUSC-MailScanner-Information: Please contact postmstr-at-musc.edu for more information 6, 27 -- X-MUSC-MailScanner: Found to be clean 6, 27 -- X-MUSC-MailScanner-SpamCheck: 6, 27 -- X-MailScanner-From: smythen-at-musc.edu 6, 27 -- Content-Transfer-Encoding: 8bit 6, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2FEGaph024908 ==============================End of - Headers==============================
} -----Original Message----- } From: biology-at-ucla.edu [mailto:biology-at-ucla.edu] } Sent: Tuesday, March 14, 2006 9:02 PM } To: Dusevich, Vladimir } Subject: [Microscopy] viaWWW: Cleaning Grids } } } } } -------------------------------------------------------------- } -------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } -------------- } } This Question/Comment was submitted to the Microscopy } Listserver using the WWW based Form at } http://www.microscopy.com/MLFormMail.html } -------------------------------------------------------------- } ------------- } Remember this posting is most likely not from a Subscriber, } so when replying } please copy both biology-at-ucla.edu as well as the } MIcroscopy Listserver } -------------------------------------------------------------- } ------------- } } Email: biology-at-ucla.edu } Name: Eric } } Organization: UCLA Medical Center } } Title-Subject: [Filtered] Cleaning Grids } } Question: This was never a problem till about a month ago.. } } Since about a month we have been having a problem keeping the } sections stuck to the grids. The grids are cleaned in 100% } Acetone and dried. Sections are picked up either from above } or below the water. } } When the grids are stained using the microwave staining } method we have been using for several years the sections come } off the grids... Every now and then the sections will stick } to the grids and everyhting is fine. } } Any suggestions about consistency? I have tried staining } less time in the microwave, but this does not make a difference... } } Any different ways to clean the grids so the sections will } stick better? } } } } -------------------------------------------------------------- } ------------- } } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Tue Mar 14 20:49:33 } 2006 12, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 12, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k2F2nV3U009322 } 12, 12 -- for {microscopy-at-microscopy.com} ; Tue, 14 Mar } 2006 20:49:31 -0600 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p0611040ac03d311d0ae9-at-[206.69.208.22]} } 12, 12 -- Date: Tue, 14 Mar 2006 20:49:31 -0600 12, 12 -- To: } microscopy-at-microscopy.com 12, 12 -- From: biology-at-ucla.edu } (by way of MicroscopyListserver) 12, 12 -- Subject: viaWWW: } Cleaning Grids 12, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== }
==============================Original Headers============================== 6, 23 -- From DusevichV-at-umkc.edu Wed Mar 15 11:16:20 2006 6, 23 -- Received: from KC-MSXPROTO2.kc.umkc.edu (pop3.exchange.umkc.edu [134.193.143.155] (may be forged)) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FHGJE7019813 6, 23 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 11:16:20 -0600 6, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by KC-MSXPROTO2.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Wed, 15 Mar 2006 11:16:19 -0600 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: RE: [Microscopy] viaWWW: Cleaning Grids 6, 23 -- Date: Wed, 15 Mar 2006 11:16:18 -0600 6, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADBFC-at-KC-MSX1.kc.umkc.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [Microscopy] viaWWW: Cleaning Grids 6, 23 -- Thread-Index: AcZH3OYO5tPkLjyjS3SyB39oOjdAHAAdviOA 6, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 6, 23 -- To: {biology-at-ucla.edu} , {microscopy-at-msa.microscopy.com} 6, 23 -- X-OriginalArrivalTime: 15 Mar 2006 17:16:19.0395 (UTC) FILETIME=[2CDFB930:01C64854] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2FHGJE7019813 ==============================End of - Headers==============================
Using pre-embedding histochemistry, a client has infiltrated lung tissue with gold nanoparticles of various sizes. He would like to see the distribution of the particles on semithin sections and then examine the tissue with TEM. Does anybody out there have a good protocol for silver-intensifying the gold on 1 micron Epon sections for light microscopy? Any suggestions would be greatly appreciated.
Ralph Common Division of Human Pathology Michigan State University
==============================Original Headers============================== 3, 24 -- From rcommon-at-msu.edu Wed Mar 15 11:57:31 2006 3, 24 -- Received: from sys10.mail.msu.edu (sys10.mail.msu.edu [35.9.75.110]) 3, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FHvTK5027212 3, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 11:57:29 -0600 3, 24 -- Received: from [35.9.122.125] (helo=emlab) 3, 24 -- by sys10.mail.msu.edu with esmtpsa (Exim 4.52 #1) 3, 24 -- (TLSv1:RC4-MD5:128) 3, 24 -- id 1FJaFV-000205-4K 3, 24 -- for Microscopy-at-microscopy.com; Wed, 15 Mar 2006 12:57:29 -0500 3, 24 -- From: "Ralph Common" {rcommon-at-msu.edu} 3, 24 -- To: {Microscopy-at-microscopy.com} 3, 24 -- Subject: Nanogold in semithin sections 3, 24 -- Date: Wed, 15 Mar 2006 12:58:12 -0500 3, 24 -- Message-ID: {002f01c6485a$07a86210$7d7a0923-at-msu.edu} 3, 24 -- MIME-Version: 1.0 3, 24 -- Content-Type: text/plain; 3, 24 -- charset="iso-8859-1" 3, 24 -- Content-Transfer-Encoding: 7bit 3, 24 -- X-Priority: 3 (Normal) 3, 24 -- X-MSMail-Priority: Normal 3, 24 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) 3, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 3, 24 -- Importance: Normal 3, 24 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
Some brands of grids are routinely treated to prevent surface oxidation. If that treatment was not carried out properly on a particular batch of grids then sections and support films tend not to stick to the grids.
So long as its presence is not a problem for any reason you can dip the grids in a solution of Poly-L-Lysine which, when dry, helps the adhesion of sections. The Poly-L-Lysine is available from most EM supplies vendors.
Good luck
Ted
--- biology-at-ucla.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the } Microscopy Listserver } using the WWW based Form at } http://www.microscopy.com/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a } Subscriber, so when replying } please copy both biology-at-ucla.edu as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: biology-at-ucla.edu } Name: Eric } } Organization: UCLA Medical Center } } Title-Subject: [Filtered] Cleaning Grids } } Question: This was never a problem till about a } month ago.. } } Since about a month we have been having a problem } keeping the sections stuck to the grids. The grids } are cleaned in 100% Acetone and dried. Sections are } picked up either from above or below the water. } } When the grids are stained using the microwave } staining method we have been using for several years } the sections come off the grids... Every now and } then the sections will stick to the grids and } everyhting is fine. } } Any suggestions about consistency? I have tried } staining less time in the microwave, but this does } not make a difference... } } Any different ways to clean the grids so the } sections will stick better? } } } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Tue Mar 14 } 20:49:33 2006 } 12, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 12, 12 -- by ns.microscopy.com (8.12.11/8.12.8) } with ESMTP id k2F2nV3U009322 } 12, 12 -- for {microscopy-at-microscopy.com} ; Tue, 14 } Mar 2006 20:49:31 -0600 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: } {p0611040ac03d311d0ae9-at-[206.69.208.22]} } 12, 12 -- Date: Tue, 14 Mar 2006 20:49:31 -0600 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: biology-at-ucla.edu (by way of } MicroscopyListserver) } 12, 12 -- Subject: viaWWW: Cleaning Grids } 12, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 10, 20 -- From drteddunne-at-yahoo.com Wed Mar 15 13:05:40 2006 10, 20 -- Received: from web33405.mail.mud.yahoo.com (web33405.mail.mud.yahoo.com [68.142.206.137]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2FJ5dDu007594 10, 20 -- for {microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 13:05:40 -0600 10, 20 -- Received: (qmail 21325 invoked by uid 60001); 15 Mar 2006 19:05:39 -0000 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 20 -- s=s1024; d=yahoo.com; 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 10, 20 -- b=QXIk89ojBUVCxHfMwJgmXomQT+Y47b0oP71D9CPlBsjufU0n8Yq6kF5JupBwUr7rh3YQepYAThkxF+WzP0fAeTi0nwJ0YJUCCt4wUs+Oe+Ca+WoL9EW1TDW3Qhzy4Ovvx473f2eeI3GKS3n48YYQlZwdmBh/XHad1gHeFAD0Xt8= ; 10, 20 -- Message-ID: {20060315190539.21323.qmail-at-web33405.mail.mud.yahoo.com} 10, 20 -- Received: from [202.47.247.116] by web33405.mail.mud.yahoo.com via HTTP; Wed, 15 Mar 2006 11:05:39 PST 10, 20 -- Date: Wed, 15 Mar 2006 11:05:39 -0800 (PST) 10, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 20 -- Subject: Re: [Microscopy] viaWWW: Cleaning Grids 10, 20 -- To: biology-at-ucla.edu 10, 20 -- Cc: microscopy-at-microscopy.com 10, 20 -- In-Reply-To: {200603150350.k2F3oXiX028013-at-ns.microscopy.com} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 10, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Hi Guys, I have a tantalum substrate covered with 30-40 nm tantalum oxide and I want to etch patterns through tantalum oxide to tantalum using HF. I am thinking to use EBL and PMMA as a photoresist, do you think PMMA stand against HF etching or the HF will etch thwe whole thing?
Thanks in advance
-- ********************************************************** Hany Ramadan Graduate student Chemistry department McMaster university, Hamilton, Ontario, Canada 905-525-9140 x: 26322 elsayeh-at-mcmaster.ca **********************************************************
==============================Original Headers============================== 4, 23 -- From ramadanhany-at-gmail.com Wed Mar 15 13:06:51 2006 4, 23 -- Received: from zproxy.gmail.com (zproxy.gmail.com [64.233.162.192]) 4, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FJ6oBo007819 4, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 13:06:50 -0600 4, 23 -- Received: by zproxy.gmail.com with SMTP id m22so195042nzf 4, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 11:06:50 -0800 (PST) 4, 23 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 23 -- s=beta; d=gmail.com; 4, 23 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 23 -- b=NHkumNmslSI2zJldfvRP8c2GKRkxSrY0kAWTX444AdD3MGrrkrsoQyVY2l+h+mVxsG0wAMLqJxrFRAOZkwnVIcT91TvoWR12EKJFdYDcOEOucEqh9GoAUDupNsQgrXawESptccR9mMjv/Y57xnvb0rFWOFTPipv8ziJOhU+Dpk0= 4, 23 -- Received: by 10.37.20.43 with SMTP id x43mr423587nzi; 4, 23 -- Wed, 15 Mar 2006 11:06:50 -0800 (PST) 4, 23 -- Received: by 10.37.12.3 with HTTP; Wed, 15 Mar 2006 11:06:50 -0800 (PST) 4, 23 -- Message-ID: {8d8ce5a30603151106v1b1f49a9x69073941151588a0-at-mail.gmail.com} 4, 23 -- Date: Wed, 15 Mar 2006 14:06:50 -0500 4, 23 -- From: "Hany Ramadan" {ramadanhany-at-gmail.com} 4, 23 -- To: Microscopy-at-microscopy.com 4, 23 -- Subject: Tantlum oxide etching aginst photoresist material 4, 23 -- MIME-Version: 1.0 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1 4, 23 -- Content-Disposition: inline 4, 23 -- Content-Transfer-Encoding: 8bit 4, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2FJ6oBo007819 ==============================End of - Headers==============================
By an interesting coincidence, the March issue of Microscopy Today has an article entitled "A Comparison of Photomicrographs Imaged Through a Late 18th C. Thomas Ribright, Cuff-Type, Brass Microscope and a Modern Olympus Optical Microscope", By D.Jones* and J.Reid.** *Retired Research Microbiologist Aberdeen and **School of Physics, The University, Aberdeen, Scot