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, Scotland. Apparently the microscope was manufactured in the late 1700's and was bought at auction recently. The microscope came in its original packing case and was accompanied by a box of accessories, which included some micro-slides by Dancer, ca 1850-60 as described in Morris' post. Jones and Reid took micrographs for the article with an Olympus OM10 single reflex 35mm camera, with lens removed and two extension tubes (20mm and 12mm) attached, fitted to the eyepiece of the microscope and held in place with a tripod stand; the source of light was a 60 watt tungsten electric light bulb in an angle-poise lamp stand. Fujichrome Professional 64T color film was used to record images. The MT article contains images of two Dancer slides, one of a very youthful Queen Victoria and the Prince Regent and another one of Trafalgar Square. No dates are given for the Dancer slides. The authors give the following reference, which may be of interest to anyone interested in Dancer photomicrographs, etc.: "Bracegirdle, B. and McCormick, J. B. The Microscopic Photographs of J.B.Dancer. Science Heritage Limited, Chicago, Illinois, 1993."
Ron Anderson, Editor Microscopy Today
keith.morris-at-ucl.ac.uk 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 } ---------------------------------------------------------------------------- } } 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 } } Tel: 020 7608 4050 } Fax: 020 7608 4034 } email: keith.morris-at-ucl.ac.uk } } } ==============================Original Headers============================== } 9, 27 -- From keith.morris-at-ucl.ac.uk Wed Mar 15 08:04:56 2006 } 9, 27 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) } 9, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FE4qu7021334 } 9, 27 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 08:04:54 -0600 } 9, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) } 9, 27 -- by vscani-d.ucl.ac.uk with smtp (Exim 4.51) } 9, 27 -- id 1FJWcM-0005uL-7N } 9, 27 -- for Microscopy-at-microscopy.com; Wed, 15 Mar 2006 14:04:50 +0000 } 9, 27 -- Message-ID: {00be01c64839$5109a480$7b865290-at-keithhigrade} } 9, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} } 9, 27 -- To: {Microscopy-at-microscopy.com} } 9, 27 -- Subject: [Microscopy] Fw: First photomicrograph } 9, 27 -- Date: Wed, 15 Mar 2006 14:04:03 -0000 } 9, 27 -- MIME-Version: 1.0 } 9, 27 -- Content-Type: text/plain; } 9, 27 -- format=flowed; } 9, 27 -- charset="iso-8859-1"; } 9, 27 -- reply-type=original } 9, 27 -- Content-Transfer-Encoding: 7bit } 9, 27 -- X-Priority: 3 } 9, 27 -- X-MSMail-Priority: Normal } 9, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 } 9, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 } 9, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information } 9, 27 -- X-UCL-MailScanner: Found to be clean } 9, 27 -- X-UCL-MailScanner-SpamCheck: } 9, 27 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 19 -- From microscopytoday-at-tampabay.rr.com Wed Mar 15 15:40:44 2006 5, 19 -- Received: from ms-smtp-02.tampabay.rr.com (ms-smtp-02-smtplb.tampabay.rr.com [65.32.5.132]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FLehGL029059 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 15 Mar 2006 15:40:43 -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-02.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k2FLebi2014810; 5, 19 -- Wed, 15 Mar 2006 16:40:42 -0500 (EST) 5, 19 -- Message-ID: {441889D3.5070106-at-tampabay.rr.com} 5, 19 -- Date: Wed, 15 Mar 2006 16:40:35 -0500 5, 19 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: keith.morris-at-ucl.ac.uk, Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] Fw: First photomicrograph 5, 19 -- References: {200603151407.k2FE7CX1022076-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200603151407.k2FE7CX1022076-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==============================
Hi all, I just had a request to use our TEM to look at the protozoan Toxoplasma. The samples are unfixed - just dried onto formvar-coated grids. Is this a safe procedure? I'm use to all samples being fixed.
Any advice would be greatly appreciated. best, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Enhancement for LM has been done for decades, both with published recipes (Danscher, Burry, Bienz and Springall and Lackie from the Hammersmith EM research group in London) as well as with commercial reagents. There are a number of companies who produce silver (and gold) enhancement reagents for light and for electron microscopy. If the particles are sufficiently enhanced you will be able to pick up individual particles in the light microscope bright field image, and certainly in epi-polarisation mode. In our experience enhancement will mostly be limited to particles on or close to the surface: they have to be exposed to become enhanced. Depending on the resin there seems to be some penetration, however, with larger particles on the surface, smaller ones below the surface. To visualise the particles in LM the particles need to be relatively big, and visualising the same specimen in EM might not be ideal. But I guess your client wants to check the specimens in LM and if a signal is found, look at unenhanced ones in EM? I initially (probably mistakenly) assumed the study was about discriminating between particle sizes after enhancement. Even though the size of the enhanced particles will somewhat depend on the initial size of the gold particles, I seriously doubt it would be possible to discriminate between sizes using LM techniques. In fact that may even be hard in electron microscopy, unless the initial particles were significantly different in size. On the other hand, double labelling using silver enhancement and ultra small gold particles has been successfully done in pre-embedding EM (Yi, H., J. L.M. Leunissen, G.-M. Shi, C.-A. Gutekunst, and S. M. Hersch. A Novel Procedure for Pre-embedding Double Immunogold-Silver Labeling at the Ultrastructural Level; J. Histochem. Cytochem., March 1, 2001; 49(3): 279 - 284)
Should you require more info, please feel welcome to contact me off- list.
Jan Leunissen
On 16/03/2006, at 7:01 AM, rcommon-at-msu.edu wrote: } } 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
Aurion - President Present Address: Costerweg 5 EM-Unit 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4797109 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://ocem.otago.ac.nz ------------------------------------------------------------------------ --------
==============================Original Headers============================== 12, 23 -- From leunissen-at-aurion.nl Wed Mar 15 19:18:43 2006 12, 23 -- Received: from mta201-rme.xtra.co.nz (mta201-rme.xtra.co.nz [210.86.15.144]) 12, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2G1IfkU018685 12, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 19:18:42 -0600 12, 23 -- Received: from mta2-rme.xtra.co.nz ([210.86.15.192]) 12, 23 -- by mta201-rme.xtra.co.nz with ESMTP 12, 23 -- id {20060316011839.QSSP29457.mta201-rme.xtra.co.nz-at-mta2-rme.xtra.co.nz} 12, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Mar 2006 14:18:39 +1300 12, 23 -- Received: from [192.168.1.20] ([222.153.174.203]) by mta2-rme.xtra.co.nz 12, 23 -- with ESMTP 12, 23 -- id {20060316011839.SKM18564.mta2-rme.xtra.co.nz-at-[192.168.1.20]} 12, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Mar 2006 14:18:39 +1300 12, 23 -- Mime-Version: 1.0 (Apple Message framework v746.2) 12, 23 -- In-Reply-To: {200603151801.k2FI1QUS028241-at-ns.microscopy.com} 12, 23 -- References: {200603151801.k2FI1QUS028241-at-ns.microscopy.com} 12, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 23 -- Message-Id: {FE407FAA-B7FD-47F4-947A-E6FE60EF3B0C-at-aurion.nl} 12, 23 -- Content-Transfer-Encoding: 7bit 12, 23 -- From: Jan Leunissen {leunissen-at-aurion.nl} 12, 23 -- Subject: Re: [Microscopy] Nanogold in semithin sections 12, 23 -- Date: Thu, 16 Mar 2006 14:18:38 +1300 12, 23 -- To: Microscopy-at-microscopy.com 12, 23 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
On Mar 14, 2006, at 6:50 PM, pekysar-at-ucdavis.edu wrote:
} 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 } Dear Pat, First, you need to be sure that you are using a pre-specimen shutter, and that it is open only during the time the image is being recorded and not when the CCD read-out is happening. This will minimize dose to the specimen. Second, when setting up the LowDose parameters, set the focus offset so that there is no overlap of the beam in focus state with the area of the CCD image in exposure state--this will, of course, vary with the magnifications in the two states. Note that this also requires that the beam size in the focus state is only a little larger than the size of the image in that state. This is quite simple to achieve on the Tecnai T12 (which we have), but may be more difficult on the CM120 (with which I have no experience). 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 15 19:36:29 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 k2G1aSLc021949 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 19:36:29 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP 4, 22 -- id 2461A3621F; Wed, 15 Mar 2006 17:36:28 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP 4, 22 -- id 41F3810A892; Wed, 15 Mar 2006 17:36:27 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603150250.k2F2oMSS009582-at-ns.microscopy.com} 4, 22 -- References: {200603150250.k2F2oMSS009582-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {c07ee33431c96f1f27b4a91aaf49803e-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: Low Dose TEM 4, 22 -- Date: Wed, 15 Mar 2006 17:45:12 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com, pekysar-at-ucdavis.edu 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both scott.streiker-at-udri.udayton.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: scott.streiker-at-udri.udayton.edu Name: Scott Streiker
Organization: University of Dayton Research Institute
Title-Subject: [Filtered] Protocol for TEM of Polymer Materials
Question: References, resources, protocols sought for preparation of polymer samples for TEM. Thanks
Scott Streiker wrote: ===================================================== Title-Subject: [Filtered] Protocol for TEM of Polymer Materials
Question: References, resources, protocols sought for preparation of polymer samples for TEM. ====================================================== We have found the text Polymer Microscopy by Linda C. Sawyer and David T. Grubb, ISBN: 0412257106, Springer, to be about as close to being a "bible" as one can get, especially if one is studying polymer blends and rubber modified systems. First published in 1987, and updated in 1995, the content seems to be nearly as timely today as the day it was published. I think the book is out of print but as of today, Amazon seems to have availability both new and used.
If you are looking to visualize lamellar structures in crystalline polymers, and/or the deformation of crystalline polymers, the "bible" for that part of polymer microscopy is Polymer Single Crystals by Philip H. Geil, published in 1962. No that is not a typo for the date. For that end of polymer microscopy, the book is nearly as relavent today as it was then. It is out of print and not listed on Amazon but it probably would be found in most university libraries.
Chuck
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==============================Original Headers============================== 12, 29 -- From cgarber-at-2spi.com Wed Mar 15 23:41:32 2006 12, 29 -- Received: from s-utl02-atpop.stsn.net (s-utl02-atpop.stsn.net [72.254.128.202]) 12, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2G5fVja017817 12, 29 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 23:41:31 -0600 12, 29 -- Received: from s-utl02-atpop.stsn.net ([127.0.0.1]) 12, 29 -- by s-utl02-atpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006031600413131337 12, 29 -- ; Thu, 16 Mar 2006 00:41:31 -0500 12, 29 -- X-Spam-Status: No, hits=0.0 required=9.9 12, 29 -- tests=ALL_TRUSTED: -2.4,AWL: 0.119,SARE_RECV_ADDR: 0.027 12, 29 -- X-Spam-Level: 12, 29 -- Received: from ibm1x23g2abfyg ([10.0.91.139]) 12, 29 -- by s-utl02-atpop.stsn.net; 12, 29 -- Thu, 16 Mar 2006 00:41:29 -0500 12, 29 -- Message-ID: {001201c648bc$43d80580$8b5b000a-at-ibm1x23g2abfyg} 12, 29 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 12, 29 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 12, 29 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 29 -- Cc: {scott.streiker-at-udri.udayton.edu} 12, 29 -- Subject: References for TEM of polymers 12, 29 -- Date: Thu, 16 Mar 2006 00:41:15 -0500 12, 29 -- MIME-Version: 1.0 12, 29 -- Content-Type: text/plain; 12, 29 -- charset="Windows-1252" 12, 29 -- X-Priority: 3 12, 29 -- X-MSMail-Priority: Normal 12, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 12, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 12, 29 -- Content-Transfer-Encoding: 8bit 12, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2G5fVja017817 ==============================End of - Headers==============================
I face 3 problems and I wondered if you could not solve them by the same solution, namely carbon-coating OVER the sections.
My first problem is with semi-thick sections (for tomography): they don't stick to the grids during contrasting and I loose them! I thought that perhaps carbon-coating after the grids are deposited on the grid would help keeping them on the grid without disturbing contrasting ?
My second problem is with ultra-thin sections: when I do EDX analysis (at 200 keV) on 70 nm thick sections on formvar film, they suffer much from the beam and usually I don't see anything when I pass in STEM mode because the area has been vaporized ;-) I thought that perhaps carbon-coating the contrasted sections would help disperse the energy of the beam?
My third problem deals with 50 nm sections deposited on grid without formvar, which are very unstable under 80 keV. Well I have difficulties to make formvar films which stick to the grids, they tend to disappear in the contrasting solutions. I wondered if I could not deposit 50 nm thick sections on grids without formvar and then carbon-coating them (so over the sections).
P.S: I clean the grids by sonicating in acetone.
Thanks in advance for your humble opinions.
Stephane (without "i")
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==============================Original Headers============================== 9, 18 -- From nizets2-at-yahoo.com Thu Mar 16 02:35:21 2006 9, 18 -- Received: from web37406.mail.mud.yahoo.com (web37406.mail.mud.yahoo.com [209.191.87.59]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2G8ZKvR029350 9, 18 -- for {microscopy-at-microscopy.com} ; Thu, 16 Mar 2006 02:35:20 -0600 9, 18 -- Received: (qmail 57273 invoked by uid 60001); 16 Mar 2006 08:35:20 -0000 9, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 18 -- s=s1024; d=yahoo.com; 9, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 18 -- b=CzFyOchbrHvDMLaexVt0ODAulB0nHLAicLGtaB2N5z/7ZfDs5WSrFRkOb9/RGDpsjsqJuZ5hcuQ3miYi2ppHnF7wm1oOezskOh/G6hOCIAxlXOYfX3hBwAEHacVfrbvvd1pnakzYUSW3/GEZ6l5T5zdu9VmAf36skYqowyjpObI= ; 9, 18 -- Message-ID: {20060316083520.57271.qmail-at-web37406.mail.mud.yahoo.com} 9, 18 -- Received: from [80.122.101.102] by web37406.mail.mud.yahoo.com via HTTP; Thu, 16 Mar 2006 00:35:20 PST 9, 18 -- Date: Thu, 16 Mar 2006 00:35:20 -0800 (PST) 9, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 18 -- Subject: carbon post-coating ;-) in TEM 9, 18 -- To: microscopy-at-microscopy.com 9, 18 -- MIME-Version: 1.0 9, 18 -- Content-Type: text/plain; charset=iso-8859-1 9, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Here is an article that ought to be of use to you:
Bassett, D. C., Olley, R. H. and Vaughan, A. S. (2003) Specimen Preparation for TEM of Polymers, in Pethrick, R. A. and Viney, C., Eds. Techniques in Polymer Organisation and Morphology Characterisation, chapter 3, pp. 73-110. Wiley.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
} From: scott.streiker-at-udri.udayton.edu } Reply-To: scott.streiker-at-udri.udayton.edu } To: hinmeigeng-at-hotmail.com } Subject: [Microscopy] viaWWW: Protocol for TEM of Polymer Materials } Date: Wed, 15 Mar 2006 23:00:24 -0600 } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 12, 22 -- From hinmeigeng-at-hotmail.com Thu Mar 16 02:41:15 2006 12, 22 -- Received: from hotmail.com (bay101-f32.bay101.hotmail.com [64.4.56.42]) 12, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2G8fEEE030528 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 02:41:15 -0600 12, 22 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 12, 22 -- Thu, 16 Mar 2006 00:41:14 -0800 12, 22 -- Message-ID: {BAY101-F321721DCF1014E3F03241ACAE70-at-phx.gbl} 12, 22 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 12, 22 -- Thu, 16 Mar 2006 08:41:11 GMT 12, 22 -- X-Originating-IP: [86.128.212.193] 12, 22 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 12, 22 -- X-Sender: hinmeigeng-at-hotmail.com 12, 22 -- Reply-To: R.H.Olley-at-reading.ac.uk 12, 22 -- In-Reply-To: {200603160500.k2G50OsZ010327-at-ns.microscopy.com} 12, 22 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 12, 22 -- To: scott.streiker-at-udri.udayton.edu, Microscopy-at-MSA.Microscopy.Com 12, 22 -- Cc: R.H.Olley-at-reading.ac.uk 12, 22 -- Subject: RE: [Microscopy] viaWWW: Protocol for TEM of Polymer Materials 12, 22 -- Date: Thu, 16 Mar 2006 08:41:11 +0000 12, 22 -- Mime-Version: 1.0 12, 22 -- Content-Type: text/plain; format=flowed 12, 22 -- X-OriginalArrivalTime: 16 Mar 2006 08:41:14.0275 (UTC) FILETIME=[62666730:01C648D5] ==============================End of - Headers==============================
I have done many Toxo TEMs. Drying the parasite kills it. There is no possibility of spoors or other environmentally stable forms. Unless you have a high power TEM, there will not be too much to see. Depends on what your user wants. David
On Mar 15, 2006, at 4:30 PM, beth-at-plantbio.uga.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 all, } I just had a request to use our TEM to look at the protozoan } Toxoplasma. The samples are unfixed - just dried onto formvar-coated } grids. } Is this a safe procedure? I'm use to all samples being fixed. } } Any advice would be greatly appreciated. } best, } Beth } } } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } http://www.plantbio.uga.edu/emlab } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } *********************************************************************** } * } *** } } } } ==============================Original } Headers============================== } 10, 18 -- From beth-at-plantbio.uga.edu Wed Mar 15 17:00:44 2006 } 10, 18 -- Received: from dogwood.plantbio.uga.edu } (dogwood.plantbio.uga.edu [128.192.26.2]) } 10, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k2FN0iSY007206 } 10, 18 -- for {microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 17:00:44 } -0600 } 10, 18 -- Received: from plantbio.uga.edu ([128.192.26.46]) } 10, 18 -- by dogwood.plantbio.uga.edu } 10, 18 -- (using TLSv1/SSLv3 with cipher DES-CBC3-SHA (168 bits)) } 10, 18 -- for microscopy-at-microscopy.com; } 10, 18 -- Wed, 15 Mar 2006 18:00:38 -0500 } 10, 18 -- Date: Wed, 15 Mar 2006 18:00:41 -0500 } 10, 18 -- Mime-Version: 1.0 (Apple Message framework v553) } 10, 18 -- Content-Type: text/plain; delsp=yes; charset=US-ASCII; } format=flowed } 10, 18 -- Subject: tem of toxoplasma } 10, 18 -- From: Beth Richardson {beth-at-plantbio.uga.edu} } 10, 18 -- To: microscopy microscopy {microscopy-at-microscopy.com} } 10, 18 -- Content-Transfer-Encoding: 7bit } 10, 18 -- Message-Id: } {8698B2A3-B477-11DA-830F-000393137C00-at-plantbio.uga.edu} } 10, 18 -- X-Mailer: Apple Mail (2.553) } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-Arizona.edu Thu Mar 16 09:40:38 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2GFeblW029764 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 09:40:37 -0600 5, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id F1E67D6F13D 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 08:40:36 -0700 (MST) 5, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id B80CAD708DB 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 08:40:35 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 5, 22 -- In-Reply-To: {200603152330.k2FNUjqm013179-at-ns.microscopy.com} 5, 22 -- References: {200603152330.k2FNUjqm013179-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {48b870b177732234386709311ac3c686-at-Arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-Arizona.edu} 5, 22 -- Subject: Re: [Microscopy] tem of toxoplasma 5, 22 -- Date: Thu, 16 Mar 2006 08:40:35 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.623) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
On Mar 16, 2006, at 12:36 AM, nizets2-at-yahoo.com wrote:
} I face 3 problems and I wondered if you could not } solve them by the same solution, namely carbon-coating } OVER the sections. } } My first problem is with semi-thick sections (for } tomography): they don't stick to the grids during } contrasting and I loose them! I thought that perhaps } carbon-coating after the grids are deposited on the } grid would help keeping them on the grid without } disturbing contrasting ? } } My second problem is with ultra-thin sections: when I } do EDX analysis (at 200 keV) on 70 nm thick sections } on formvar film, they suffer much from the beam and } usually I don't see anything when I pass in STEM mode } because the area has been vaporized ;-) I thought that } perhaps carbon-coating the contrasted sections would } help disperse the energy of the beam? } } My third problem deals with 50 nm sections deposited } on grid without formvar, which are very unstable under } 80 keV. Well I have difficulties to make formvar films } which stick to the grids, they tend to disappear in } the contrasting solutions. I wondered if I could not } deposit 50 nm thick sections on grids without formvar } and then carbon-coating them (so over the sections). } } P.S: I clean the grids by sonicating in acetone. } } Thanks in advance for your humble opinions. } Dear Stephane, Carbon coating might not affect your semi-thick sections coming loose during contrasting, but a previous post suggested coating the grid with polylysine before placing the sections on it, and another suggested a brief acid dip to roughen the grid surface, so I'd try these. Carbon coating semi-thick specimens, however, will aid both thermal and electrical conductivity, so I do recommend that you coat your specimens, and if you are using a formvar substrate, carbon coat that before placing the sections. From this answer, you can surmise that my answer to your second problem is to carbon coat both the formvar before placing the thin sections and carbon coat the sections after. I would suggest either acid-dipping the grids and covering them with formvar, or using a higher-mesh grid without formvar (depending on how large an unobstructed field of view you need) and carbon coat the sections to increase stability of your 50 nm sections. In the latter case I even suggest coating both sides with carbon--being careful not to dislodge the sections when you coat the underside of the grid, of course. 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 Thu Mar 16 13:06:44 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 k2GJ6hDo009886 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 16 Mar 2006 13:06:43 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 2EDE810AA79 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 16 Mar 2006 11:06:43 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 08FD710AA62 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 16 Mar 2006 11:06:42 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603160836.k2G8aGps029523-at-ns.microscopy.com} 4, 22 -- References: {200603160836.k2G8aGps029523-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {64e50971d37899b5d5bebd33aa3d09da-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] carbon post-coating ;-) in TEM 4, 22 -- Date: Thu, 16 Mar 2006 11:15:32 -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 earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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Email: scott.streiker-at-udri.udayton.edu Name: Scott Streiker
Organization: University of Dayton reserach Institute
Title-Subject: [Filtered] Polymer preparation for TEM
Question: Thanks to all who responded to question re: preparation polymer samples for TEM.
As a result of good advice I have acquired copy "Polymer Microscopy by, Sawyer and Grubb Cheers,
Scott Streiker Electron Microscopist NEST Laboratory University of Dayton Research Institute Scott.Streiker-at-udri.udayton.edu Phone: (937)229-5776
Browsing through this month's European Semiconductor this morning I find an article on page 46 beginning..
"Hillsboro (Oregon, USA) based FEI has been in the microscopy business since 1949, when it produced the world's first Transmission Electron Microscope (TEM). Today FEI manufactures a full range of microscopes..."
Now I'm no history buff, but I didn't see an FEI label on the replica of the first TEM in the Science museum in London when I visited last year.. How can they make this claim?
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Fri Mar 17 05:48:23 2006 8, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2HBmM8R011251 8, 31 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 05:48:22 -0600 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-5.tower-78.messagelabs.com!1142596059!44508703!15 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 13263 invoked from network); 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-5.tower-78.messagelabs.com with SMTP; 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 8, 31 -- Fri, 17 Mar 2006 11:48:15 +0000 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 31 -- Content-class: urn:content-classes:message 8, 31 -- MIME-Version: 1.0 8, 31 -- Content-Type: text/plain; 8, 31 -- charset="us-ascii" 8, 31 -- Subject: The first TEM 8, 31 -- Date: Fri, 17 Mar 2006 11:48:14 -0000 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047C36-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: The first TEM 8, 31 -- Thread-Index: AcZJuKzBYGI+b8m5RBapPz+el04xIw== 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 17 Mar 2006 11:48:15.0713 (UTC) FILETIME=[AD4FB510:01C649B8] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2HBmM8R011251 ==============================End of - Headers==============================
Who's first? While some might argue that it was JJ Thompson with his work on cathode ray, let us skip forward to Ernst Ruska and Max Knoll who developed the first magnetic lens electron microscope and published in 1932 (Ann. d. Physik 12, 607 (1932). In North America, J Hiller and A Prebus at the University of Toronto, built and operated the first TEM. I will never forget reading about how they dissolved rubber bands in solvent to produce vacuum greases. (I just have to squeeze a tube...) Hiller left to join RCA to manufacture the first commercial TEM in North America, the EMB. I don't have a exact date, but Hillier and Ramberg published an article about this in J. Appl. Phys., 18, 48, 1947. Meanwhile in Europe, von Borries and Ruska were developing/manufacturing an instrument similar to the EMB at Siemens-Halske and published in Z. wiss Mikroskop 56, 317 (1939).
When I started at Goodyear tire, they had an RCA EMU-3 that was just about 25 years old, placing it's purchase in 1954 and was only a few years younger then I was. I have a publication containing a line drawing of the lab's founder, Wilisfort (I'm sure that a mis-spelling) using a EBM during the second World War. I had no idea I was walk in the shadow of such illuminated history.
The question to answer is what companies evolved into FEI? If it wasn't either RCA or Siemens-Halske the claim should be rejected.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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richard.beanland-at- bookham.com To: frank.karl-at-degussa.com cc: 03/17/2006 07:02 Subject: [Microscopy] The first TEM AM Please respond to richard.beanland
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Browsing through this month's European Semiconductor this morning I find an article on page 46 beginning..
"Hillsboro (Oregon, USA) based FEI has been in the microscopy business since 1949, when it produced the world's first Transmission Electron Microscope (TEM). Today FEI manufactures a full range of microscopes..."
Now I'm no history buff, but I didn't see an FEI label on the replica of the first TEM in the Science museum in London when I visited last year.. How can they make this claim?
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Fri Mar 17 05:48:23 2006 8, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2HBmM8R011251 8, 31 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 05:48:22 -0600 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-5.tower-78.messagelabs.com!1142596059!44508703!15 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 13263 invoked from network); 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-5.tower-78.messagelabs.com with SMTP; 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 8, 31 -- Fri, 17 Mar 2006 11:48:15 +0000 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 31 -- Content-class: urn:content-classes:message 8, 31 -- MIME-Version: 1.0 8, 31 -- Content-Type: text/plain; 8, 31 -- charset="us-ascii" 8, 31 -- Subject: The first TEM 8, 31 -- Date: Fri, 17 Mar 2006 11:48:14 -0000 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047C36-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: The first TEM 8, 31 -- Thread-Index: AcZJuKzBYGI+b8m5RBapPz+el04xIw== 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 17 Mar 2006 11:48:15.0713 (UTC) FILETIME=[AD4FB510:01C649B8] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2HBmM8R011251 ==============================End of - Headers==============================
==============================Original Headers============================== 35, 18 -- From frank.karl-at-degussa.com Fri Mar 17 07:26:40 2006 35, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 35, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HDQdUk021737 35, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Mar 2006 07:26:39 -0600 35, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 35, 18 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2HDQX9L030681; 35, 18 -- Fri, 17 Mar 2006 14:26:34 +0100 35, 18 -- In-Reply-To: {200603171202.k2HC2YWs013693-at-ns.microscopy.com} 35, 18 -- Subject: Re: [Microscopy] The first TEM 35, 18 -- To: richard.beanland-at-bookham.com, microscopy-at-msa.microscopy.com 35, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 35, 18 -- Message-ID: {OF7447929D.9CF0BD6C-ON85257134.00470EE7-85257134.0049D191-at-degussa.com} 35, 18 -- From: frank.karl-at-degussa.com 35, 18 -- Date: Fri, 17 Mar 2006 08:26:17 -0500 35, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 35, 18 -- 03/17/2006 07:26:35 AM 35, 18 -- MIME-Version: 1.0 35, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Quite easily. But I think they will find if they look into their innermost souls (if I am not spawning some appalling oxymoron here) that FEI was not even a gleam in it's creator's eye in 1949. Indeed, probably FEI's creators were not even yet gleams in their parents eyes. In fact, ...... no, forget it!
*Philips* introduced a commercial TEM in 1949, but Zeiss also claim a first commercial TEM (EM7) in 1949 and JEOL's first was in 1950, but Leeds University web site reports receiving its first TEM on 6 January 1944, installed in the ladies lavatory of the Textiles laboratory! Is it not the case that the first commercial TEMs were designed by Ruska and produced by Siemens, and that several tens of instruments were out there by 1945?? Ruska and Knoll built a TEM in 1931, and Ruska later built one by himself in 1933 that bettered the resolution of light microscopes.
see http://nobelprize.org/physics/laureates/1986/ruska-autobio.html
Chris
----- Original Message ----- X-from: {richard.beanland-at-bookham.com} To: {cjeffree-at-staffmail.ed.ac.uk} Sent: Friday, March 17, 2006 11:51 AM
How can FEI claim to have "produced the world's first Transmission Electron Microscope (TEM)"?
Blowed if I know. From my understanding, it's not the first TEM (1931, Ruska and Knoll), or the first commercial TEM (1936, MetropolitanVickers EM1), or even the first commercial series of TEMs (1939, Siemens). If these don't count as TEMs in some way, I'd like to know.
I think some PR person has read a line saying "Philips first TEM" and re-written it as "The first TEM"...
Mike Fay School of Mechanical, Materials and Manufacturing Engineering Nottingham University University Park Nottingham NG7 2RD tel 0115 8466081
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Not only did FEI not "produce the world's first" TEM in 1949 -- they did not manufacture the first commercial one either.
My understanding is that the first commercial TEM was a Metropolitan- Vickers instrument manufactured in 1936 in England. It was shortly followed by a superior microscope from Siemens and Halske in 1939 in Germany. Hillier, Zworykin, and Snyder at RCA were working on electron microscopy in the 1930s and 1940s, putting out their first commercial model in 1940. In the 1940s, Hillier was also outfitting TEMs with EELS and designing the electron microprobe a few years before Castaing.
However, at the FEI website under "A Brief History of FEI", I found the statement: "1949: Philips Electron Optics introduces the world's first commercial transmission electron microscope (TEM)." I, too, am at a loss to explain their claim.
Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
On Mar 17, 2006, at 6:30 AM, richard.beanland-at-bookham.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 } ---------------------------------------------------------------------- } ------ } } Browsing through this month's European Semiconductor this morning I } find } an article on page 46 beginning.. } } "Hillsboro (Oregon, USA) based FEI has been in the microscopy business } since 1949, when it produced the world's first Transmission Electron } Microscope (TEM). Today FEI manufactures a full range of } microscopes..." } } Now I'm no history buff, but I didn't see an FEI label on the } replica of } the first TEM in the Science museum in London when I visited last } year.. } How can they make this claim? } } Richard } } ________________________________________ } Richard Beanland } Analytical Services } Bookham Inc } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } } ====================================================================== } = } This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly } prohibited. } No part of this message can be considered a request for goods or } services. } ====================================================================== } = } } } ==============================Original } Headers============================== } 8, 31 -- From richard.beanland-at-bookham.com Fri Mar 17 05:48:23 2006 } 8, 31 -- Received: from mail78.messagelabs.com } (mail78.messagelabs.com [195.245.230.131]) } 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id } k2HBmM8R011251 } 8, 31 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 } 05:48:22 -0600 } 8, 31 -- X-VirusChecked: Checked } 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com } 8, 31 -- X-Msg-Ref: server-5.tower-78.messagelabs.com!1142596059! } 44508703!15 } 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- } 8, 31 -- X-Originating-IP: [213.249.209.179] } 8, 31 -- Received: (qmail 13263 invoked from network); 17 Mar 2006 } 11:48:19 -0000 } 8, 31 -- Received: from unknown (HELO cas-smx- } brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 8, 31 -- by server-5.tower-78.messagelabs.com with SMTP; 17 Mar } 2006 11:48:19 -0000 } 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI } ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with } Microsoft SMTPSVC(6.0.3790.211); } 8, 31 -- Fri, 17 Mar 2006 11:48:15 +0000 } 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 8, 31 -- Content-class: urn:content-classes:message } 8, 31 -- MIME-Version: 1.0 } 8, 31 -- Content-Type: text/plain; } 8, 31 -- charset="us-ascii" } 8, 31 -- Subject: The first TEM } 8, 31 -- Date: Fri, 17 Mar 2006 11:48:14 -0000 } 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047C36-at-cas- } smx-02.BOOKHAM.ENTERPRISE.PRI} } 8, 31 -- X-MS-Has-Attach: } 8, 31 -- X-MS-TNEF-Correlator: } 8, 31 -- Thread-Topic: The first TEM } 8, 31 -- Thread-Index: AcZJuKzBYGI+b8m5RBapPz+el04xIw== } 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 8, 31 -- To: {microscopy-at-microscopy.com} } 8, 31 -- X-OriginalArrivalTime: 17 Mar 2006 11:48:15.0713 (UTC) } FILETIME=[AD4FB510:01C649B8] } 8, 31 -- Content-Transfer-Encoding: 8bit } 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k2HBmM8R011251 } ==============================End of - } Headers============================== }
==============================Original Headers============================== 9, 18 -- From frah0010-at-umn.edu Fri Mar 17 08:09:29 2006 9, 18 -- Received: from mtaout-a.tc.umn.edu (mtaout-a.tc.umn.edu [134.84.119.206]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HE9ScH001691 9, 18 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 08:09:28 -0600 9, 18 -- Received: from [160.94.146.133] (212-swing.geo.umn.edu [160.94.146.133]) by mtaout-a.tc.umn.edu with ESMTP; Fri, 17 Mar 2006 08:09:27 -0600 (CST) 9, 18 -- X-Umn-Remote-Mta: [N] 212-swing.geo.umn.edu [160.94.146.133] #+LO+TS+AU+HN 9, 18 -- In-Reply-To: {200603171230.k2HCUvdj018545-at-ns.microscopy.com} 9, 18 -- References: {200603171230.k2HCUvdj018545-at-ns.microscopy.com} 9, 18 -- Mime-Version: 1.0 (Apple Message framework v746.3) 9, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 18 -- Message-Id: {1F3BDA84-09EF-4A54-AEA1-2ACD89920E5D-at-umn.edu} 9, 18 -- Cc: Ellery Frahm {frah0010-at-umn.edu} 9, 18 -- Content-Transfer-Encoding: 7bit 9, 18 -- From: Ellery Frahm {frah0010-at-umn.edu} 9, 18 -- Subject: Re: [Microscopy] The first TEM 9, 18 -- Date: Fri, 17 Mar 2006 08:09:25 -0600 9, 18 -- To: Microscopy-at-microscopy.com 9, 18 -- X-Mailer: Apple Mail (2.746.3) ==============================End of - Headers==============================
You just know there's going to be a few responses to this. So mine is brief.
Yes I've got to agree. I'm sure Ernst Ruska got a Nobel Prize for it (admittedly it took about 50 years) - maybe FEI know something different. First effective design and implementation of a resolution better than a light microscope between 1931 and 1933 then with Siemens first commercial instrument 1939 - again no mention of FEI or Phillips.
I'm sure that by 1949 most industrial countries had the beginnings of commercial instruments.
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: richard.beanland-at-bookham.com
{ SEQ CHAPTER \h \r 1}Hello everybody, We want to upgrade of our digital system. We have a digital camera on our H7500. It is a Hamamatsu Advantage HR side mounted one megapixel camera. Is there an advantage of having 6 megapixel versus 4 megapixel camera for biological materials? Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Fri Mar 17 08:49:22 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HEnJ0J015311 1, 22 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 08:49:20 -0600 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1FKGGT-000613-01 1, 22 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 10:49:17 -0400 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 17 Mar 06 10:49:16 -0400 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 17 Mar 06 10:48:50 -0400 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Fri, 17 Mar 2006 10:43:53 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: TEM digital camera 1, 22 -- Message-ID: {441A92E9.14981.3CD146-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
As two of my main interests are photography and microscopes, I have had a more thorough look at the on-line archives to find out who made the first photomicrograph (it was surprisingly easy - articles on photography appear as well represented on the internet as those on the computer). If you are interested please read further :-
X-from 'offline' discussions, it seems that Fox Talbot (or possibly others such as the Rev. J.B. Meade or Daguerre) could claim to have produced the first documented 'fixed' photomicrograph (a magnified image on light sensitive paper). Fox Talbot certainly used a 'solar microscope' for his 'photogenic drawings' in the previous year to Dancer's Flea (shown in 1840). I did find a reference to the 'solar microscope' without having to visit any Museums. It's clearly essentially a camera obscura (lucida) with enlargement 'microscope' lenses to produce magnified images for tracing onto paper or exposing light sensitive paper (e.g. a photograph of 'lace' attributed to Fox Talbot in Sept 1839). I think even that counts as a photomicrograph. Problems with long-term fixing of the image means its not very impressive now. Solar microscopes had been around for a long time as an artists aid, long before Fox Talbot used it to focus images onto his light sensitive paper. They were easily available to those with the money to afford one. I have seen a well made 'solar microscope' example dating from 1750, see http://www.mhs.ox.ac.uk/cameras/index.htm?item18 - this museum in Oxford also an original 1839 Fox Talbot 'lace' photograph (image on-line).
In 1835 Talbot produced the first 'negative' image - the Oriel window in the South Gallery at Lacock Abbey, Wiltshire (near where I live - and it's where he died). This is now the oldest surviving photo (negative) using todays photographic process, although others such as Daguerre were doing similar work at the time (but his method had no means of reproducing the image via the 'negatives'). I can't quickly find any on-line images of Fox Talbot or others more scientific solar microscope 'photogenic drawings', but they are mentioned in the letter from the Rev. J. B. Reade to Fox Talbot suggesting that his photographic use of gallic acid preceded that of Talbot :
"So early as April 1839 the Rev. J.B. Reade made a sensitive paper by using infusion of galls after nitrate of silver; by this process Mr. Reade obtained several drawings of microscopic objects by means of the solar microscope; the drawings were taken before the paper was dry.".
X-from other letters it seems that Talbot had used a solar microscope for photograph enlargements (photomicrographs) before January 1839 (see below). So it's almost certain Talbot (or another early photographer) will have beaten Dancer to producing the first photomicrograph, although as Dancer's 'flea' was 'exhibited' in 1840 he may have made earlier versions. It's also likely that Daguerre and others will have produced magnified images during their experiments from 1935 to 1939, although even Daguerre had problems with fixing the image until 1837.
When Daguerre make public his photographic process on the 7th Jan 1839, Fox Talbot wrote to William Jerdan (Royal Society) on the 30th January 1839 emphasising that his early work was independent of Daguerre and was in no way influenced by Daguerre's research. Fox Talbot wrote: "The Specimens of this art which I exhibited at the Royal Institution, though consisting only of what I happened to have with me in Town, are yet sufficient to give a general idea of it, and to shew the wide range of its applicability. Among them were pictures of flowers and leaves; a pattern of lace; figures taken from painted glass; a view of Venice copied from an engraving, some images formed by the Solar Microscope, viz. a slice of wood very highly magnified, exhibiting the pores of two kinds, one set much smaller than the other, and more numerous. Another Microscopic sketch, exhibiting the reticulations on the wing of an insect. Finally: various pictures, representing the architecture of my house in the country; all these made with the Camera Obscura in the summer of 1835". This conflicts with some reports that Talbot probably purchased his 'solar microscope' in March 1839 (a receipt in his collection isn't specific on what items were bought then). A lot of Fox Talbot correspondence is now online at http://www.foxtalbot.arts.gla.ac.uk.
However, one of the first photographs that Daguerre displayed publicly was of a dead spider seen in the solar microscope. This was cited in a 6th January 1839 letter from H. Gaucheraud that had originally appeared in La Gazette de France; Fox Talbot likely became aware of this when the letter was reprinted in The Literary Gazette, on the 12 January 1839. So the first photomicrograph is about as old as the first photograph. I expect they were all rushing about then looking for novel images to make into photographs - or 'photogenic drawings' as they called them, e.g. Talbot mentions in a letter (2nd Nov 1839) "I saw Cooper at the Polytechnic taking a drawing with the Oxy hydrogen Microscope* in three minutes". *A microscope employing the light produced by the burning of lime under a current of oxyhydrogen gas.
Also Niépce, Daguerre's early collaborator, is universally credited with producing the first successful photograph (heliograph) in June/July 1827. Besides even the image on the medieval Turin shroud appears to be similar to a 'photographic' one - although it may not have been created intentionally. Also many were experimenting before the 1827 date. Indeed Sir Humphrey Davy (1778-1829) the chemist, together with Thomas Wedgwood (1771-1805), son of Josiah Wedgwood the potter, undertook photographic experiments. In 1800 Davy and Wedgwood succeeded in taking a photograph, they did not, however succeed in fixing the image. Indeed years later, Fox Talbot remembered that "the first person who applied photography to the solar microscope was undoubtedly Mr Wedgwood - but none of his delineations have been preserved, and I believe that no particulars are known. Next in order of time to Mr Wedgwood's, came my own experiments. Having published my first photographic process in January, 1839, I immediately applied it to the solar microscope, and in the course of that year made a great many microscopic photographs, which I gave away to Sir John Herschell, Sir Walter Calverley Trevelyan, and other friends". In Sept 8th 1839 Talbot wrote "I have great hopes of this branch of the Art proving very useful, as for instance in copying the forms of minute crystallization which are so complicated as almost to defy the pencil" - he clearly also had an interest in taking photographs of crystals.
So there are many investigators that could have got there first, although most likely their methods of image capture didn't have the 'negative' reproduction capabilities of Talbot's technique and/or their photo-image just faded soon after capture.
If you are interested in more microscope 'victoriana' also have a look at http://www.diatoms.co.uk where Klaus Kemp has turned diatoms (and butterfly scales) back into an art form.
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 Tel: 020 7608 4050 Fax: 020 7608 4034 email: keith.morris-at-ucl.ac.uk
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Maybe this is pie in the sky, but perhaps, just perhaps, someone from FEI might address this issue, if they have the courage.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: malcolm.haswell-at-sunderland.ac.uk [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Friday, March 17, 2006 10:37 AM To: kenconverse-at-qualityimages.biz
Richard
You just know there's going to be a few responses to this. So mine is brief.
Yes I've got to agree. I'm sure Ernst Ruska got a Nobel Prize for it (admittedly it took about 50 years) - maybe FEI know something different. First effective design and implementation of a resolution better than a light microscope between 1931 and 1933 then with Siemens first commercial instrument 1939 - again no mention of FEI or Phillips.
I'm sure that by 1949 most industrial countries had the beginnings of commercial instruments.
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: richard.beanland-at-bookham.com
Hi, Ralph
The new NTegra Tomo provides an interesting solution. It combines AFM directly into a Leica microtome so that you can do sections of an appropriate depth, but use the AFM for imaging. We have used this sort of system very successfully already for imaging, sizing and counting hard particles in polymer matrices (silica in PS/HIPS blend). The differences in the local elasticity will image the lung tissue without having to enhance the nanoparticles and, clearly, the difference in hardness will image the nanoparticles. We can have samples run if you would like.
Contact me off-line for further details.
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
CAVEAT: MME is involved in the support of this product.
P. S. Need a good general reference or light microscopy text for the Spring semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.
At 12:26 PM 3/15/2006, rcommon-at-msu.edu wrote:
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==============================Original Headers============================== 16, 17 -- From bfoster-at-mme1.com Fri Mar 17 12:36:04 2006 16, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 16, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HIa31g003592 16, 17 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 12:36:03 -0600 16, 17 -- Received: (qmail 25814 invoked from network); 17 Mar 2006 13:37:37 -0500 16, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 16, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 17 Mar 2006 13:37:37 -0500 16, 17 -- Message-Id: {7.0.1.0.0.20060317123205.01ffece0-at-mme1.com} 16, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 16, 17 -- Date: Fri, 17 Mar 2006 12:36:08 -0600 16, 17 -- To: rcommon-at-msu.edu, Microscopy Listserver {microscopy-at-microscopy.com} 16, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 16, 17 -- Subject: Re: [Microscopy] Nanogold in semithin sections 16, 17 -- In-Reply-To: {200603151826.k2FIQXtO001828-at-ns.microscopy.com} 16, 17 -- References: {200603151826.k2FIQXtO001828-at-ns.microscopy.com} 16, 17 -- Mime-Version: 1.0 16, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
The April, 2003, issue of Microscopy and Microanalysis contains an article, "Key Events in the History of Electron Microscopy" which states that in 1937, "The Metropolitan Vickers Company (Manchester, UK) supplies the first commercial electron microscope to Louis C. Martin at Imperial College, London". By way of trivia - The B.F. Goodrich Company (Akron, Ohio) purchased a Philips EM100B in 1957, the delivery of which was conditional upon the approval of W. Ladd whom BFG hired to check out the instrument which was assembled for just that purpose at the Philips' facilities in Mt. Vernon, NY. My...how times have changed!
Ron Smith, Lake Havasu City, AZ
==============================Original Headers============================== 3, 17 -- From Ron2450-at-aol.com Fri Mar 17 12:36:42 2006 3, 17 -- Received: from imo-m28.mx.aol.com (imo-m28.mx.aol.com [64.12.137.9]) 3, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HIae6V003713 3, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 12:36:41 -0600 3, 17 -- Received: from Ron2450-at-aol.com 3, 17 -- by imo-m28.mx.aol.com (mail_out_v38_r7.3.) id w.2a4.76b7015 (17079) 3, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 13:36:34 -0500 (EST) 3, 17 -- From: Ron2450-at-aol.com 3, 17 -- Message-ID: {2a4.76b7015.314c5bb1-at-aol.com} 3, 17 -- Date: Fri, 17 Mar 2006 13:36:33 EST 3, 17 -- Subject: Re: First Commercial TEM 3, 17 -- To: Microscopy-at-microscopy.com 3, 17 -- MIME-Version: 1.0 3, 17 -- Content-Type: text/plain; charset="US-ASCII" 3, 17 -- Content-Transfer-Encoding: 7bit 3, 17 -- X-Mailer: 9.0 SE for Windows sub 5022 3, 17 -- X-Spam-Flag: NO ==============================End of - Headers==============================
Hi group - this is what I found when I did a Google search: Early experiments using X-rays of extremely short wavelength were not pursued further because of the inability to focus these rays. The first breakthrough in the development of the electron microscope came when Louis de Broglie advanced his theory that the electron had a dual nature, with characteristics of a particle or a wave. The demonstration, in 1923 by Busch, that a beam of electrons could be focused by magnetic or electric fields opened the way for the development of the first electron microscope, in 1932, by Knoll and Ruska. Although the initial development of the electron microscope, in Germany, was followed by technical improvements in America, the first commercially available apparatus was marketed by Seimens.
==============================Original Headers============================== 1, 18 -- From maloneyb-at-fiu.edu Fri Mar 17 15:40:58 2006 1, 18 -- Received: from smtp10.fiu.edu (smtp10.fiu.edu [131.94.79.27]) 1, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HLevp5024909 1, 18 -- for {microscopy-at-ns.microscopy.com} ; Fri, 17 Mar 2006 15:40:57 -0600 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu [131.94.95.156]) 1, 18 -- by smtp10.fiu.edu (MOS 3.7.1-GA) 1, 18 -- with ESMTP id CLW05883; 1, 18 -- Fri, 17 Mar 2006 16:40:56 -0500 (EST) 1, 18 -- Message-ID: {441B2B93.6060308-at-fiu.edu} 1, 18 -- Date: Fri, 17 Mar 2006 16:35:15 -0500 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 18 -- X-Accept-Language: en-us, en 1, 18 -- MIME-Version: 1.0 1, 18 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-ns.microscopy.com} 1, 18 -- Subject: first TEM 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Contrary to common impression, it appears Ruska and Knoll were not even aware of de Broglie's work when they started trying to build an electron microscope, in fact they were initially not happy to hear about it because they had hoped to escape the limits imposed by wave optics! In his Nobel lecture in 1988 Ruska said:
"As engineers we did not know yet the thesis on the "material waves" by French physicist de Broglie that had been put forward several years earlier (1925). . . When I first heard of it in summer 1931, I was very much disappointed that the resolution should now be limited again by a wavelength (of the materiestrahlung). I was immediately heartened, though, when with the aid of the de Broglie equation I became satisfied that these waves must be around five orders of magnitude shorter in length than light waves. Thus, there was no reason to abandon the aim of electron microscopy surpassing the resolution of light microscopy".
Other interesting facts can be found in "EMSA and Its people the First Fifty Years" by Sterling Newberry and published by the Electron Microscopy Society of America in 1992.
Marie
On Mar 17, 2006, at 5:12 PM, maloneyb-at-fiu.edu wrote:
} Early experiments using X-rays of extremely short wavelength were not } pursued further because of the inability to focus these rays. The first } breakthrough in the development of the electron microscope came when } Louis de Broglie advanced his theory that the electron had a dual } nature, with characteristics of a particle or a wave. The } demonstration, } in 1923 by Busch, that a beam of electrons could be focused by magnetic } or electric fields opened the way for the development of the first } electron microscope, in 1932, by Knoll and Ruska. Although the initial } development of the electron microscope, in Germany, was followed by } technical improvements in America, the first commercially available } apparatus was marketed by Seimens. } } ==============================Original } Headers============================== } 1, 18 -- From maloneyb-at-fiu.edu Fri Mar 17 15:40:58 2006 } 1, 18 -- Received: from smtp10.fiu.edu (smtp10.fiu.edu [131.94.79.27]) } 1, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k2HLevp5024909 } 1, 18 -- for {microscopy-at-ns.microscopy.com} ; Fri, 17 Mar 2006 } 15:40:57 -0600 } 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu } [131.94.95.156]) } 1, 18 -- by smtp10.fiu.edu (MOS 3.7.1-GA) } 1, 18 -- with ESMTP id CLW05883; } 1, 18 -- Fri, 17 Mar 2006 16:40:56 -0500 (EST) } 1, 18 -- Message-ID: {441B2B93.6060308-at-fiu.edu} } 1, 18 -- Date: Fri, 17 Mar 2006 16:35:15 -0500 } 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} } 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; } rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) } 1, 18 -- X-Accept-Language: en-us, en } 1, 18 -- MIME-Version: 1.0 } 1, 18 -- To: "'microscopy-at-msa.microscopy.com'" } {microscopy-at-ns.microscopy.com} } 1, 18 -- Subject: first TEM } 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed } 1, 18 -- Content-Transfer-Encoding: 7bit } ==============================End of - } Headers============================== } } Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 8, 21 -- From marie.cantino-at-uconn.edu Sat Mar 18 09:07:48 2006 8, 21 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 8, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2IF7lbP015649 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 09:07:48 -0600 8, 21 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 8, 21 -- by mail1.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2IF7eK11604 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 10:07:40 -0500 8, 21 -- Mime-Version: 1.0 (Apple Message framework v623) 8, 21 -- In-Reply-To: {200603172212.k2HMClqn030823-at-ns.microscopy.com} 8, 21 -- References: {200603172212.k2HMClqn030823-at-ns.microscopy.com} 8, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 8, 21 -- Message-Id: {e5a8d5e1aab9c76980512131f1fbaff6-at-uconn.edu} 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 8, 21 -- Subject: Re: [Microscopy] first TEM 8, 21 -- Date: Sat, 18 Mar 2006 10:07:40 -0500 8, 21 -- To: Microscopy-at-Microscopy.Com 8, 21 -- X-Mailer: Apple Mail (2.623) 8, 21 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 8, 21 -- X-UConn-MailScanner: Found to be clean 8, 21 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
Has anyone else had problems in negative staining using recent bottles of uranyl acetate? None of our newer bottles (we have purchased several from several different vendors in the past year) work as well as one purchased in 1991. Problems include poor spreading and precipitation or aggregation of the stain. The newer stuff is much more soluble than the old, has a strong odor of acetic acid, and is lighter in color. Some of the new bottles work better than others (though none as well as the old stuff), but there was no correlation with any variable except pH. Lots that worked best had higher pH in solution (3.5-3.8) than poor lots (2.3-2.5), but raising the pH of the solution did not produce better staining. Results were quite variable between lots from the same vendor. As far as I can tell, all lots work OK for positive staining of resin sections (though we don't want to waste any of the "best" bottle making tests on sections!)
We're wondering whether anyone else has noticed this and found a better source or a way to improve the existing lots.
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 5, 19 -- From marie.cantino-at-uconn.edu Sat Mar 18 09:37:07 2006 5, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2IFb6KM021362 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 09:37:06 -0600 5, 19 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 5, 19 -- by mail2.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2IFb1l15528 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 10:37:01 -0500 5, 19 -- Mime-Version: 1.0 (Apple Message framework v623) 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- Message-Id: {d84f87dd1f62e47529f153e4ea711649-at-uconn.edu} 5, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 5, 19 -- To: Microscopy-at-Microscopy.Com 5, 19 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 5, 19 -- Subject: Uranyl acetate/negative staining problems 5, 19 -- Date: Sat, 18 Mar 2006 10:37:00 -0500 5, 19 -- X-Mailer: Apple Mail (2.623) 5, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 5, 19 -- X-UConn-MailScanner: Found to be clean 5, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
I'm not sure if you got your question answered yet.
Here are a few questions for you to help fill in some gaps.
What feature size(s) are you dealing with? Why are you using a Ta substrate?
I suspect that any TaN etchant will also etch the Ta. Thus, there is no stop mechanism for etching just the TaN.
TaN is sometimes put on top of Cu damascene runners. These are plasma etched using an oxidizing plasma. If you want to use wet etch, I would change the substrate to something that is not going to be etched, or coat whatever you have with Si3N4 which is a stop. Then, use Transene Etch III.
If your feature sizes are too small, wet etch won't do a good job, IMO. If the resist is thick enough and cured, any type should work. Which type you use is going to depend on UV wavelength and what is used to pattern/expose the resist as well as compatable developer.
gary g.
At 11:34 AM 3/15/2006, you wrote:
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==============================Original Headers============================== 13, 21 -- From gary-at-gaugler.com Sat Mar 18 12:04:42 2006 13, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2II4eGY004349 13, 21 -- for {microscopy-at-microscopy.com} ; Sat, 18 Mar 2006 12:04:41 -0600 13, 21 -- Received: (qmail 4270 invoked from network); 18 Mar 2006 10:04:38 -0800 13, 21 -- Received: by simscan 1.1.0 ppid: 4267, pid: 4268, t: 0.1675s 13, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 21 -- by qsmtp3 with SMTP; 18 Mar 2006 10:04:38 -0800 13, 21 -- Message-Id: {6.2.3.4.2.20060318095716.0205d038-at-mail.calweb.com} 13, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 13, 21 -- Date: Sat, 18 Mar 2006 10:04:39 -0800 13, 21 -- To: ramadanhany-at-gmail.com 13, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 21 -- Subject: Re: [Microscopy] Tantlum oxide etching aginst photoresist 13, 21 -- material 13, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 21 -- In-Reply-To: {200603151934.k2FJYCTe014754-at-ns.microscopy.com} 13, 21 -- References: {200603151934.k2FJYCTe014754-at-ns.microscopy.com} 13, 21 -- Mime-Version: 1.0 13, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Um... might be safer to say North America than just America, if it is Hillier's work you're thinking of. The group he was in made a practical TEM at the University of Toronto in 1938 (if I recall correctly), and he didn't move to the US until 1940.
Mike Fay School of Mechanical, Materials and Manufacturing Engineering Nottingham University University Park Nottingham NG7 2RD tel 0115 8466081
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Hi group - this is what I found when I did a Google search: Early experiments using X-rays of extremely short wavelength were not pursued further because of the inability to focus these rays. The first breakthrough in the development of the electron microscope came when Louis de Broglie advanced his theory that the electron had a dual nature, with characteristics of a particle or a wave. The demonstration, in 1923 by Busch, that a beam of electrons could be focused by magnetic or electric fields opened the way for the development of the first electron microscope, in 1932, by Knoll and Ruska. Although the initial development of the electron microscope, in Germany, was followed by technical improvements in America, the first commercially available apparatus was marketed by Seimens.
==============================Original Headers============================== 1, 18 -- From maloneyb-at-fiu.edu Fri Mar 17 15:40:58 2006 1, 18 -- Received: from smtp10.fiu.edu (smtp10.fiu.edu [131.94.79.27]) 1, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HLevp5024909 1, 18 -- for {microscopy-at-ns.microscopy.com} ; Fri, 17 Mar 2006 15:40:57 -0600 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu [131.94.95.156]) 1, 18 -- by smtp10.fiu.edu (MOS 3.7.1-GA) 1, 18 -- with ESMTP id CLW05883; 1, 18 -- Fri, 17 Mar 2006 16:40:56 -0500 (EST) 1, 18 -- Message-ID: {441B2B93.6060308-at-fiu.edu} 1, 18 -- Date: Fri, 17 Mar 2006 16:35:15 -0500 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 18 -- X-Accept-Language: en-us, en 1, 18 -- MIME-Version: 1.0 1, 18 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-ns.microscopy.com} 1, 18 -- Subject: first TEM 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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==============================Original Headers============================== 13, 24 -- From michael.fay-at-nottingham.ac.uk Mon Mar 20 04:57:24 2006 13, 24 -- Received: from haydn.is.nottingham.ac.uk (haydn.is.nottingham.ac.uk [128.243.40.92]) 13, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KAvN7J001360 13, 24 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 04:57:23 -0600 13, 24 -- Received: from ccw0m1.ccc.nottingham.ac.uk ([128.243.220.65] helo=ccw0m1.nottingham.ac.uk) 13, 24 -- by haydn.is.nottingham.ac.uk with esmtp (Exim 3.36 #2) 13, 24 -- id 1FLI4X-0002DA-00 13, 24 -- for microscopy-at-microscopy.com; Mon, 20 Mar 2006 10:57:13 +0000 13, 24 -- Received: from Gwweb1-MTA by ccw0m1.nottingham.ac.uk 13, 24 -- with Novell_GroupWise; Mon, 20 Mar 2006 10:57:14 +0000 13, 24 -- Message-Id: {s41e8a8a.006-at-ccw0m1.nottingham.ac.uk} 13, 24 -- X-Mailer: Novell GroupWise Internet Agent 6.0.3 13, 24 -- Date: Mon, 20 Mar 2006 10:57:01 +0000 13, 24 -- From: "Michael Fay" {Michael.Fay-at-nottingham.ac.uk} 13, 24 -- To: {microscopy-at-microscopy.com} 13, 24 -- Subject: Re: [Microscopy] first TEM 13, 24 -- Mime-Version: 1.0 13, 24 -- Content-Type: text/plain; charset=US-ASCII 13, 24 -- Content-Disposition: inline 13, 24 -- X-MailScanner-Information: Please contact staff-it-helpline-at-nottingham.ac.uk for more information 13, 24 -- X-MailScanner: Found to be clean 13, 24 -- X-MailScanner-From: michael.fay-at-nottingham.ac.uk 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2KAvN7J001360 ==============================End of - Headers==============================
Sorry to bother the list with this, but one person responding to my question about UA left a phone message for me, which I accidentally deleted before writing down the name and correct phone number. If you are that person, could you please call or e-mail again? Thanks!
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 4, 19 -- From marie.cantino-at-uconn.edu Mon Mar 20 10:20:11 2006 4, 19 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KGKAjZ016191 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 10:20:11 -0600 4, 19 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 4, 19 -- by mail1.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2KGA0K14987 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- Message-Id: {f8d53214ee4a81c62c984e77fbf3b665-at-uconn.edu} 4, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 19 -- To: microscopy-at-Microscopy.Com 4, 19 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 4, 19 -- Subject: Uranyl acetate/negative staining problems 4, 19 -- Date: Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- X-Mailer: Apple Mail (2.623) 4, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 4, 19 -- X-UConn-MailScanner: Found to be clean 4, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (danielluth-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 16, 2006 at 19:01:41 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both danielluth-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: danielluth-at-yahoo.com Name: Daniel Luth
Organization: University of Melbourne
Education: Graduate College
Location: Melbourne, Victoria, Australia
Title: Are parasites detectable in the blood from a microscope?
Question: Hi, My wife recently went to a naturopath who had a degree in Microscopy, and took a sample of her blood to view under the microscope. The image was projected onto a monitor. The naturopath was able to detect parasites in the blood - and pointed them out. My wife described them as wiggly moving threads. My question is are parasites detectable in the blood from a microscope?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ml687644-at-bigpond.net.au) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 17, 2006 at 19:28:50 ---------------------------------------------------------------------------
Email: ml687644-at-bigpond.net.au Name: Mark LESTER
Organization: New South Wales Police Service
Education: Undergraduate College
Location: Sydney, Australia
Question: Hi there. I am trying to find out the technique you would use for optimum visualisation and identification of the following specimens;
a)bullet for rifling marks b)sperm cells c)hair d)natural and synthetic fibres e)flies f)spiracles on maggots g)paint flakes (for counting pain layers to trace paint source) h)cannabis leaf to identify cystolithic hairs i)vomit for identifying food particles j)a soil sample k)gunshot residue
if you could outline the optical system used and the appropriate magnification, that would be of great assistance to me.
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 julie.duimstra-at-hti.hutch.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Replica Materials other than Cellulose Acetate for SEM Analysis of Metal Surfaces
Question: What recommendations would you have for replicating materials, other than cellulose acetate, for making replicas of flat metal surfaces that would be used for subsequent viewing and EDS analysis with SEM?
I am interested in any techniques that you might be using, literature citations and/or vendors of replicating materials other than cellulose acetate for materials and biological applications.
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Email: amich-at-ufl.edu Name: Albina
Organization: UF
Title-Subject: [Filtered] vapor fixation
Question: Hi, I would appreciate your suggestions on the following: I work on the project studying bacterial attachment/interaction with various substrates. Some of those substrates are polymers and fibers which cannot withstand standard fixation by immersion since in some cases I would like to avoid wetting the fibers (they swell). I wanted to try vapor fixation; but since it pilot project I am reluctant to pump osmium vapor with pump. Can anyone suggest a way to vapor-fix samples with homemade ìon-the-cheapî setup? It is also my understanding that I would need to vapor fix with osmium first and follow with glutaraldehyde, right? Thank you for consideration, Albina
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Email: b.ambrose-at-massey.ac.nz Name: Barbara Ambrose
Organization: Massey University
Title-Subject: [Filtered] The Ideal Microscopy Center
Question: Dear All We have just secured a large grant to fund a microscopy suite (MMIC) here in New Zealand. MMIC would house a spinning disc confocal microscope, eSEM, TEM, light microscopy and all the processing equipment. We are having to renovate a new space for the microscopy suite and am wondering if you have any advice/suggestions or if you could redesign your suite what you would incorporate or change. Thank you for your time, cheers barbara
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Email: pekysar-at-ucdavis.edu Name: Pat Kysar
Organization: University of California, Davis
Title-Subject: [Filtered] Thanks for Low Dose Info
Question: Thank you to all who responded to my questions about low dose TEM! I have some very good ideas/suggestions. We are very lucky to have such a great resouce at our disposal! Pat Kysar University of California, Davis Medical School, Pathology EM Lab
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Email: Paul.Leahy-at-epa.vic.gov.au Name: Paul Leahy
Organization: EPA Victoria
Title-Subject: [Filtered] Field Microscopes
Question: I'm after a field microscope for looking at small algae. I've come accross the Swift FM-31 and one from Richardson Technologies. Can anyone comment on these? Also does anyone know of suppliers in Australia?
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Email: Paul.Leahy-at-epa.vic.gov.au Name: Paul Leahy
Organization: EPA Victoria
Title-Subject: [Filtered] Field Microscopes
Question: I'm after a field microscope for looking at small algae. I've come accross the Swift FM-31 and one from Richardson Technologies. Can anyone comment on these? Also does anyone know of suppliers in Australia?
Sorry to bother the list with this, but one person responding to my question about UA left a phone message for me, which I accidentally deleted before writing down the name and correct phone number. If you are that person, could you please call or e-mail again? Thanks!
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 4, 19 -- From marie.cantino-at-uconn.edu Mon Mar 20 11:29:22 2006 4, 19 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KGKAjZ016191 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 10:20:11 -0600 4, 19 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 4, 19 -- by mail1.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2KGA0K14987 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- Message-Id: {f8d53214ee4a81c62c984e77fbf3b665-at-uconn.edu} 4, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 19 -- To: microscopy-at-microscopy.com 4, 19 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 4, 19 -- Subject: Uranyl acetate/negative staining problems 4, 19 -- Date: Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- X-Mailer: Apple Mail (2.623) 4, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 4, 19 -- X-UConn-MailScanner: Found to be clean 4, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
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Email: pekysar-at-ucdavis.edu Name: Pat Kysar
Organization: University of California, Davis
Title-Subject: [Filtered] Thanks for Low Dose Info
Question: Thank you to all who responded to my questions about low dose TEM! I have some very good ideas/suggestions. We are very lucky to have such a great resouce at our disposal! Pat Kysar University of California, Davis Medical School, Pathology EM Lab
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 b.ambrose-at-massey.ac.nz as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: b.ambrose-at-massey.ac.nz Name: Barbara Ambrose
Organization: Massey University
Title-Subject: [Filtered] The Ideal Microscopy Center
Question: Dear All We have just secured a large grant to fund a microscopy suite (MMIC) here in New Zealand. MMIC would house a spinning disc confocal microscope, eSEM, TEM, light microscopy and all the processing equipment. We are having to renovate a new space for the microscopy suite and am wondering if you have any advice/suggestions or if you could redesign your suite what you would incorporate or change. Thank you for your time, cheers barbara
Out of fairness, I wanted to point out FEI has changed its website to reflect our discussion here:
On Friday it read:
"1949: Philips Electron Optics introduces the world's first commercial transmission electron microscope (TEM)."
Today it reads:
"1949: Philips Electron Optics (part of FEI Company since 1997) was one of the first companies to start volume production of Transmission Electron Microscopes in 1949."
Thumbs up to FEI for changing their website so quickly when the error was pointed out.
Best, Ellery
-------------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
==============================Original Headers============================== 8, 17 -- From frah0010-at-umn.edu Mon Mar 20 11:40:13 2006 8, 17 -- Received: from mtaout-w.tc.umn.edu (mtaout-w.tc.umn.edu [160.94.160.21]) 8, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KHeDUd009714 8, 17 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 11:40:13 -0600 8, 17 -- Received: from [160.94.146.133] (212-swing.geo.umn.edu [160.94.146.133]) by mtaout-w.tc.umn.edu with ESMTP for Microscopy-at-microscopy.com; Mon, 20 Mar 2006 11:39:20 -0600 (CST) 8, 17 -- X-Umn-Remote-Mta: [N] 212-swing.geo.umn.edu [160.94.146.133] #+LO+TS+AU+HN 8, 17 -- Mime-Version: 1.0 (Apple Message framework v746.3) 8, 17 -- In-Reply-To: {200603171731.k2HHVe4l032728-at-ns.microscopy.com} 8, 17 -- References: {200603171731.k2HHVe4l032728-at-ns.microscopy.com} 8, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 17 -- Message-Id: {6A92218F-31CC-4C03-B890-00A51AB2043D-at-umn.edu} 8, 17 -- Content-Transfer-Encoding: 7bit 8, 17 -- From: Ellery Frahm {frah0010-at-umn.edu} 8, 17 -- Subject: Re: [Microscopy] The first TEM 8, 17 -- Date: Mon, 20 Mar 2006 11:39:17 -0600 8, 17 -- To: Microscopy-at-microscopy.com 8, 17 -- X-Mailer: Apple Mail (2.746.3) ==============================End of - Headers==============================
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Title-Subject: [Filtered] Replica Materials other than Cellulose Acetate for SEM Analysis of Metal Surfaces
Question: What recommendations would you have for replicating materials, other than cellulose acetate, for making replicas of flat metal surfaces that would be used for subsequent viewing and EDS analysis with SEM?
I am interested in any techniques that you might be using, literature citations and/or vendors of replicating materials other than cellulose acetate for materials and biological applications.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ml687644-at-bigpond.net.au) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 17, 2006 at 19:28:50 ---------------------------------------------------------------------------
Email: ml687644-at-bigpond.net.au Name: Mark LESTER
Organization: New South Wales Police Service
Education: Undergraduate College
Location: Sydney, Australia
Question: Hi there. I am trying to find out the technique you would use for optimum visualisation and identification of the following specimens;
a)bullet for rifling marks b)sperm cells c)hair d)natural and synthetic fibres e)flies f)spiracles on maggots g)paint flakes (for counting pain layers to trace paint source) h)cannabis leaf to identify cystolithic hairs i)vomit for identifying food particles j)a soil sample k)gunshot residue
if you could outline the optical system used and the appropriate magnification, that would be of great assistance to me.
This Question was submitted to Ask-A-Microscopist by (danielluth-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 16, 2006 at 19:01:41 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both danielluth-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: danielluth-at-yahoo.com Name: Daniel Luth
Organization: University of Melbourne
Education: Graduate College
Location: Melbourne, Victoria, Australia
Title: Are parasites detectable in the blood from a microscope?
Question: Hi, My wife recently went to a naturopath who had a degree in Microscopy, and took a sample of her blood to view under the microscope. The image was projected onto a monitor. The naturopath was able to detect parasites in the blood - and pointed them out. My wife described them as wiggly moving threads. My question is are parasites detectable in the blood from a microscope?
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Email: amich-at-ufl.edu Name: Albina
Organization: UF
Title-Subject: [Filtered] vapor fixation
Question: Hi, I would appreciate your suggestions on the following: I work on the project studying bacterial attachment/interaction with various substrates. Some of those substrates are polymers and fibers which cannot withstand standard fixation by immersion since in some cases I would like to avoid wetting the fibers (they swell). I wanted to try vapor fixation; but since it pilot project I am reluctant to pump osmium vapor with pump. Can anyone suggest a way to vapor-fix samples with homemade ìon-the-cheapî setup? It is also my understanding that I would need to vapor fix with osmium first and follow with glutaraldehyde, right? Thank you for consideration, Albina
African trypanosomes would be detectable with a light microscope and would look like worms moving between the red blood cells, but I don't think sleeping sickness has reached Australia yet.
You could probably detect Giardia in unstained specimens but they are round cells.
What was the final diagnosis and treatment/
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
Email: danielluth-at-yahoo.com Name: Daniel Luth
Organization: University of Melbourne
Education: Graduate College
Location: Melbourne, Victoria, Australia
Title: Are parasites detectable in the blood from a microscope?
Question: Hi, My wife recently went to a naturopath who had a degree in Microscopy, and took a sample of her blood to view under the microscope. The image was projected onto a monitor. The naturopath was able to detect parasites in the blood - and pointed them out. My wife described them as wiggly moving threads. My question is are parasites detectable in the blood from a microscope?
==============================Original Headers============================== 16, 22 -- From PWebster-at-hei.org Mon Mar 20 11:58:03 2006 16, 22 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 16, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KHw3IG028454 16, 22 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 11:58:03 -0600 16, 22 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 16, 22 -- Mon, 20 Mar 2006 17:58:02 +0000 16, 22 -- User-Agent: Microsoft-Entourage/11.2.1.051004 16, 22 -- Date: Mon, 20 Mar 2006 09:58:00 -0800 16, 22 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: parasites 16, 22 -- detectable in the blood 16, 22 -- From: "Webster, Paul" {PWebster-at-hei.org} 16, 22 -- To: {danielluth-at-yahoo.com} 16, 22 -- CC: {microscopy-at-microscopy.com} 16, 22 -- Message-ID: {C0442D28.8E6E%PWebster-at-hei.org} 16, 22 -- Thread-Topic: [Microscopy] [Filtered] AskAMicroscopist: parasites 16, 22 -- detectable in the blood 16, 22 -- Thread-Index: AcZMR9NqEcoBzrg7EdqcYAANk7Zh7g== 16, 22 -- In-Reply-To: {200603201753.k2KHrSUE017918-at-ns.microscopy.com} 16, 22 -- Mime-version: 1.0 16, 22 -- Content-type: text/plain; 16, 22 -- charset="US-ASCII" 16, 22 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
Yes, there are parasites that show up on blood films, usually with a special stain. Live wiggling things might easily be white blood cells doing their normal amoboid movement or just brownian motion. I for one would not trust my diagnosis to a "naturopath who had a degree in Microscopy". This sounds like quackery to me. There is a school of (what I consider charlatans) practioners that diagnose everything from blood smears. If your wife has a parasitic infection she will have distinct symptoms. Do you have objections to physicians with real medical training?
Geoff
danielluth-at-yahoo.com 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 **********************************************
==============================Original Headers============================== 7, 32 -- From mcauliff-at-umdnj.edu Mon Mar 20 11:58:44 2006 7, 32 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 7, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KHwhIP030774 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 11:58:44 -0600 7, 32 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 7, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 9192A4BE1D 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 12:58:43 -0500 (EST) 7, 32 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 7, 32 -- by zix03.umdnj.edu (Proprietary) with ESMTP id D9EC54BE03 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 12:58:42 -0500 (EST) 7, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 7, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 7, 32 -- id {0IWF00L01T3NCO-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 7, 32 -- for microscopy-at-msa.microscopy.com; Mon, 20 Mar 2006 12:58:42 -0500 (EST) 7, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 7, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 7, 32 -- 2004)) with ESMTP id {0IWF00AV7TXAGV-at-Polaris.umdnj.edu} ; Mon, 7, 32 -- 20 Mar 2006 12:58:23 -0500 (EST) 7, 32 -- Date: Mon, 20 Mar 2006 12:57:27 -0500 7, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 7, 32 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: parasites detectable in 7, 32 -- the blood 7, 32 -- In-reply-to: {200603201705.k2KH5IXo029145-at-ns.microscopy.com} 7, 32 -- To: danielluth-at-yahoo.com, MicroscopyListserver {microscopy-at-msa.microscopy.com} 7, 32 -- Message-id: {441EED07.2080201-at-umdnj.edu} 7, 32 -- MIME-version: 1.0 7, 32 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 7, 32 -- Content-transfer-encoding: 7BIT 7, 32 -- X-Accept-Language: en-us, en 7, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 7, 32 -- Gecko/20040804 Netscape/7.2 (ax) 7, 32 -- References: {200603201705.k2KH5IXo029145-at-ns.microscopy.com} ==============================End of - Headers==============================
Julie Duimstra wrote: =========================================================== Title-Subject: [Filtered] Replica Materials other than Cellulose Acetate for SEM Analysis of Metal Surfaces
Question: What recommendations would you have for replicating materials, other than cellulose acetate, for making replicas of flat metal surfaces that would be used for subsequent viewing and EDS analysis with SEM?
I am interested in any techniques that you might be using, literature citations and/or vendors of replicating materials other than cellulose acetate for materials and biological applications =========================================================== Could you tell us what it is that you find objectionable to cellulose acetate? That information would be helpful in determining that might be a menu of possible alternatives. Normally on a metal surface, CA works just fine.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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==============================Original Headers============================== 12, 25 -- From cgarber-at-2spi.com Mon Mar 20 13:09:37 2006 12, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 12, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KJ9buN017167 12, 25 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 13:09:37 -0600 12, 25 -- Received: from ibm1x23g2abfyg ([70.5.4.200]) 12, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k2KJ8wrV032355; 12, 25 -- Mon, 20 Mar 2006 14:09:03 -0500 12, 25 -- X-IDV-FirstRcvd: [70.5.4.200] 12, 25 -- X-IDV-HELO: ibm1x23g2abfyg 12, 25 -- Message-ID: {006501c64c51$bef55950$c8040546-at-ibm1x23g2abfyg} 12, 25 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 12, 25 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 12, 25 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 25 -- Cc: {julie.duimstra-at-hti.hutch.com} 12, 25 -- Subject: Alternatives to cellulose acetate as a replicating resin 12, 25 -- Date: Mon, 20 Mar 2006 14:08:15 -0500 12, 25 -- MIME-Version: 1.0 12, 25 -- Content-Type: text/plain; 12, 25 -- charset="Windows-1252" 12, 25 -- X-Priority: 3 12, 25 -- X-MSMail-Priority: Normal 12, 25 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 12, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 12, 25 -- Content-Transfer-Encoding: 8bit 12, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2KJ9buN017167 ==============================End of - Headers==============================
You don't give us the reason you are looking for an alternative to the acetate replicas, or much detail about what you are trying to do.
You talk about EDS analysis. Are you perhaps trying to extract particles from the surface and analyze them? If so, then a very common technique 20 years ago was the carbon extraction replica. This involves lightly etching a surface to get the particles standing proud, then evaporating a layer of carbon, and finally a deeper etch to release the particles - now embedded in the carbon film, which is floated off on the surface of water. The film is picked up on a grid and examined in the TEM or SEM - depending on the particles. For applications where carbon is not appropriate, other materials can be used (to my knowledge, Al and amorphous SiO have been used, and possibly others).
In a variation (dating to even earlier in the history of microscopy), the replica would have a light "shadow" of a metal like Pt evaporated on it at an oblique angle, to enhance visualization of the topography of the surface. This was often used to allow surface topography to be investigated in the TEM, before SEMs were common (which, I hasten to add, was before I was active in the field!)
If this is anything like what you have in mind, I would be happy to expand on the procedure off-line.
Tony Garratt-Reed.
At 12:46 PM 3/20/2006, you wrote:
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*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
You could use vapors from warm or hot paraformaldehyde (the solid) or acrolein (a liquid) or osmium. Note that osmium can be dissolved in solvents other than water. Or perhaps follow one of the first two with osmium for lipid staining. You don't have to pump osmiun, just put the object of interest in a closed container with the fixative of your choice, the put the container under a fume hood.
Geoff
amich-at-ufl.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 **********************************************
==============================Original Headers============================== 8, 31 -- From mcauliff-at-umdnj.edu Mon Mar 20 15:23:14 2006 8, 31 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KLNEF2006472 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 15:23:14 -0600 8, 31 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 8, 31 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 1A2AE4BE66 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 15:23:14 -0600 (CST) 8, 31 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 8, 31 -- by zix02.umdnj.edu (Proprietary) with ESMTP id D30CE4BE38 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 15:23:12 -0600 (CST) 8, 31 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 31 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 31 -- id {0IWG00H012D4JC-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 31 -- for microscopy-at-msa.microscopy.com; Mon, 20 Mar 2006 16:23:12 -0500 (EST) 8, 31 -- Received: from [127.0.0.1] ([10.138.2.240]) 8, 31 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 31 -- 2004)) with ESMTP id {0IWG00BM23EDKD-at-Polaris.umdnj.edu} ; Mon, 8, 31 -- 20 Mar 2006 16:23:01 -0500 (EST) 8, 31 -- Date: Mon, 20 Mar 2006 16:22:06 -0500 8, 31 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 31 -- Subject: Re: [Microscopy] viaWWW: vapor fixation 8, 31 -- In-reply-to: {200603201709.k2KH9SMU030393-at-ns.microscopy.com} 8, 31 -- To: amich-at-ufl.edu, MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 31 -- Message-id: {441F1CFE.2030102-at-umdnj.edu} 8, 31 -- MIME-version: 1.0 8, 31 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 8, 31 -- Content-transfer-encoding: 8BIT 8, 31 -- X-Accept-Language: en-us, en 8, 31 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 31 -- Gecko/20040804 Netscape/7.2 (ax) 8, 31 -- References: {200603201709.k2KH9SMU030393-at-ns.microscopy.com} ==============================End of - Headers==============================
Hi Barbara, There should be several responses concerning analytical labs in the archives for the listserver as this topic always becomes important when one has to build a lab. The archives can be checked at the listserver address. Also, I am sending you off line (so attachments can be included) one of my articles and checklists for designing analytical facilities. If you have any questions, please let me know. In my many years in microscopy, that has been the part that has been the most fun i.e. designing labs and I have been lucky enough to design many of them and all of them definitely represent a different challenge. Best of Luck,
Judy
Judy Murphy, PhD Microscopy Training, Imaging, and Lab Design Stockton, CA 95219 murphyjudy-at-comcast.net
b.ambrose-at-massey.ac.nz wrote:
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==============================Original Headers============================== 8, 19 -- From murphyjudy-at-comcast.net Mon Mar 20 16:21:20 2006 8, 19 -- Received: from sccrmhc14.comcast.net (sccrmhc14.comcast.net [63.240.77.84]) 8, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KMLKAf016913 8, 19 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 16:21:20 -0600 8, 19 -- Received: from [192.168.1.101] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 8, 19 -- by comcast.net (sccrmhc14) with ESMTP 8, 19 -- id {2006032022211801400c881me} ; Mon, 20 Mar 2006 22:21:19 +0000 8, 19 -- Message-ID: {441F2AE3.6030105-at-comcast.net} 8, 19 -- Date: Mon, 20 Mar 2006 14:21:23 -0800 8, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 8, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 8, 19 -- X-Accept-Language: en-us, en 8, 19 -- MIME-Version: 1.0 8, 19 -- To: b.ambrose-at-massey.ac.nz, Microscopy List Server {microscopy-at-microscopy.com} 8, 19 -- Subject: Re: [Microscopy] viaWWW: The Ideal Microscopy Center? 8, 19 -- References: {200603201739.k2KHdvnA009000-at-ns.microscopy.com} 8, 19 -- In-Reply-To: {200603201739.k2KHdvnA009000-at-ns.microscopy.com} 8, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
We are going to add a Hitachi S-2700 SEM to our lab but it is missing the instruction book and diagrams. I have a request in to Hitachi to see if they can come up with anything, but just in case, anyone have a set to share? Maybe I can copy them and get them back to you ASAP.
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============================== 4, 21 -- From jmkrupp-at-ucsc.edu Mon Mar 20 16:53:14 2006 4, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 4, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KMrDwb026767 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 16:53:13 -0600 4, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 4, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.1/8.13.1) with ESMTP id k2KMloEZ023619 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 14:47:58 -0800 (PST) 4, 21 -- Received: from [128.114.25.214] (account jmkrupp-at-ucsc.edu HELO [128.114.25.197]) 4, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 4, 21 -- with ESMTPA id 49810387 for microscopy-at-microscopy.com; Mon, 20 Mar 2006 14:47:49 -0800 4, 21 -- Mime-Version: 1.0 4, 21 -- Message-Id: {p06230907c044e0eacbff-at-[128.114.25.197]} 4, 21 -- Date: Mon, 20 Mar 2006 14:47:45 -0800 4, 21 -- To: microscopy-at-microscopy.com 4, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 4, 21 -- Subject: Hitachi S-2700 instructions 4, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 4, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 4, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 4, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (csucla-at-charter.net) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, March 20, 2006 at 13:40:39 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both csucla-at-charter.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: csucla-at-charter.net Name: Sandra Brodwin
Organization: Liebman & Associates
Education: Graduate College
Location: La Canada, California, USA
Title: work situation of a microscopist
Question: I am a rehabilitation consultant. I have been asked to evaluate the worksite of a microscopist who has bilateral carpel tunnel for suggestions on how to change her work situation to allow her to continue to do the work in her field. Therefore, I need to know 1) what to look for in her work station and 2) idea on how to change/adapt the work station to accommodate bilateral carpal tunnel syndrome.
You don't say whether you are talking about light microscopy or some form of electron microscopy or something else.
I would guess that it would be absolutely imperative to find out how the sufferer works with the instrument(s) in question. It sounds as if they may be using rotary controls to shift specimens around repeatedly and possibly the layout of the instrument/workstation doesn't encourage a sensible close and comfortable seating position. These are certainly problems I have heard of when someone sits at a scanning electron microscope and needs to examine lots of areas on a specimen and I would suspect that their may be similar problems with many light microscope specimen stages.
I think that the problem is that many manufacturers did not envisage operators sitting at their microscopes for 6 or 7 hours and constantly rotating the control knobs. But I suppose that it may happen in medical screening or quality control where repetitive work loads can be high.
Some manufacturers may provide more ergonomic control systems, work planning or furniture might be re-arranged. But it's difficult to suggest more without some detail.
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: csucla-at-charter.net
(Posting for a colleague in Bethesda,MD)
Department of Health and Human Services
National Institutes of Health, USA
National Heart, Lung and Blood Institute
Biologist (Electron Microscopy)
The Electron Microscopy Core Facility of the Genetics and Development Biology Center, Division of Intramural Research in the National Heart, Lung and Blood Institute, is recruiting for a Biologist (Electron Microscopy). To be minimally qualified you must have a bachelor's degree in biology or related field. Experience is required in performing fixation, embedding and ultra-thin sectioning of samples for transmission electron microscopy as well as operating the electron microscope to record images. Experience with ultra-thin cryo-microtomy and immunogold labeling is highly desirable. Other desirable areas of experience include rotary shadowing, freeze fracture, operation of the scanning electron microscope and SEM specimen preparation. The successful candidate will be hired at a level and with salary compensation commensurate with previous experience and qualifications. The appointee must be a US citizen. For more information or to apply, please reference vacancy announcement number NHLBI-06-114086 and the website, WWW.USAJOBS.GOV {http://www.usajobs.gov/} {http://www.usajobs.gov/} .
The NIH is an Equal Opportunity Employer. Applications from women, minorities, and persons with disabilities are strongly encouraged. The NHLBI/NIH is a smoke-free environment.
This email and its attachments are intended for the use of the individual or entity who is the intended recipient and may contain information that is privileged, confidential and exempt from disclosure or any type of use under applicable law. If the reader of this email is not the intended recipient, or the employee, agent or representative responsible for delivering the email to the intended recipient, you are hereby notified that any dissemination, distribution, copying or other use of this email is strictly prohibited. If you have received this email in error, please reply immediately to the sender. MA
Malcolm's reply has covered the microscopes. However, modern optical and SEM microscopes invariably have a computer and mouse attached to them these days, or the user then goes back to the office PC, so CTS is a very common problem. I use mouse controlled PC's all day for microscope image capture and control, associated office and image analysis work, and at home for fun throughout the night. I also play with my young son on the Playstation (Tekken kick boxing in particular is a killer for the wrist). So naturally I get mild CTS occasionally - a clear sign to use the mouse less for a few days, switch to a racing sim, and so rest the wrist for a while.
Try using the keyboard (e.g. for shortcuts) if you have real problems. Also you can get a Cirque Smartcat laptop style large USB touchpad to replace the mouse (for about £60) if you have limited movement - or at least get a decent optical mouse - I prefer the MS optical Intellimouse although I disable the extra side buttons. I've never tried the Smartcat, as it will be a bit slower than a mouse, although it did receive a very good review (www.pcpro.co.uk). I have tried wrist supports (with gels etc.) but I found them very uncomfortable and stick to resting my wrist on the benchtop. A visit to my chiropractor occasionally does help me (joint manipulation and firing a little captive bolt thingy at the affected wrist bones) and for convenience I use a freeze gel rather than icepacks to reduce inflammation (so I can still use the mouse during the 'physiotherapy'). Arranging the workstation helps with neck pain but does not really do anything for my CTS, although wrist angle is supposedly important. I never find the microscope focus knob a problem at all - suppose most of ours are motorised and so have far less resistance than manual focus gears. On all our mechanical stages the manual XY stage control is on the left side, and so won't irritate the same wrist (the right hand is used for the focus).
UCl have standard health & safety handouts for workstation setup similar to http://www.afscme.org/health/faq-cts.htm but offer us no real specific advice on CTS
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: {csucla-at-charter.net} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, March 21, 2006 12:25 AM
Forgot to add the link, but also have a look at
http://www.safecomputing.com
for other PC based anti-RSI ideas.
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: {csucla-at-charter.net} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, March 21, 2006 12:25 AM
Position available: Professor in Electron Microscopy and Director of the Center for Electron Nanoscopy, Technical University of Denmark Application deadline: 2 May 2006
The Technical University of Denmark (DTU) announces a position as full professor in Electron Microscopy to be filled as soon as possible. The professor will lead research in Electron Microscopy and at the same time serve as Director for our new Center for Electron Nanoscopy (CEN).
We are in the process of establishing an advanced center for electron microscopy, which will complement our strong activities within nanotechnology and materials science. The center, which is made possible by a generous donation by the A.P. Moller Foundation, will be equipped with a total of six microscopes of which two will be absolute state of the art TEM's with sub-Ångstrom resolution, energy loss spectroscopy, and 3D structural analysis facilities. Moreover, one of the TEM's will be modified into an environmental TEM. CEN is expected to be fully operational in the fall of 2007.
We seek a dynamic person, who will carry out advanced research in Electron Microscopy and at the same time direct activities in CEN. In the start-up phase of CEN we expect the candidate to interact with equipment manufacturers, to coordinate hiring of additional staff and to overlook the construction of a new building for CEN. When CEN is established the candidate will be responsible for the daily operation, the portfolio of research projects and the services offered, assisted by two senior scientists and several technicians. CEN will interact with a number of DTU departments within the fields related to nanotechnology including metallurgy, polymers and advanced materials, catalysis, solid state physics, micro- and nano-electronics, optics and photonics. Moreover, CEN will interact with outside partners from academia and industry. It will be the responsibility of the candidate to ensure that CEN has the capability to carry out both advanced research projects as well as general analysis. In the capacity of Director the candidate reports to a Board under the Rector of DTU.
We seek a candidate with the highest academic qualifications and demonstrated abilities in leading advanced research at an international level related to electron microscopy. We envision a background in physics or engineering with expertise in the fields of nanotechnology, materials science or chemistry and with the following qualifications:
· A strong international research reputation with demonstrated ability in leading large scale projects and securing major research funding.
· Outstanding expertise in the use of electron microscopy as part of advanced research within materials science/nanotechnology
· Ability to manage and strengthen a team of scientists and technicians
· Flexibility in assuring a balance between advanced research projects and general type service jobs in the center.
· Ability to collaborate actively with many research groups in different fields and to initiate new contacts.
We expect the candidate to participate actively in teaching, development of new courses in electron microscopy as well as to supervise students both at M.Sc. and Ph.D. levels. Teaching experience at university level in an international student environment is therefore considered an advantage.
The salary and appointment terms will be negotiated in accordance with the current collective agreement for Danish University faculty members.
All interested candidates irrespective of age, gender, race, religion or ethnic background are invited to apply.
Applications should include a detailed resumé with a list of publications, a statement of teaching and research interests as well as visions and plans for the future development of the Center of Nanoscopy. Copies of key publications for assessment of the research experience should be included in the application that should be sent to Rector Lars Pallesen, Building 101 A, Technical University of Denmark, DK-2800 Lyngby, Denmark.
For more information contact Dean of Research, Prof. Kristian Stubkjaer (forskningsdekan-at-adm.dtu.dk, direct telephone +45 4525 1008).
The application with enclosures in triplicate must be received before 2 May 2006 at 12:00.
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 nicol-at-semiconductor.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: nicol-at-semiconductor.com Name: Nick Aitken
Title-Subject: [Filtered] sputter coaters
Question: Hi all,
we are currently looking at replacing our old Hummer X coater. It is not producing coatings that look good enough at high magnification. We are currently looking at a couple of turbo pumped coaters by various manufacturers, but I thought I would get some external opinions. First of all our requirements: We need a good conductive coating that will not show a lot of grain at around 250 thousand times mag : repeatability is a must. We have multiple users preparing many samples and we need consistent results across the board
What manufacturers should we be looking at, and what target material would be the best for this type of use? We would be coating semiconductor devices in both topographical and cross sectional orientations.
Hi folks, CCD camera technology has moved on a lot since I bought one for our optical microscopes back in 1998. The camera still works fine, but it has always struggled with low light levels (on-chip integration being an expensive option in those days). Does anyone have any recommendations for a relatively cheap camera (+ controller/software + framegrabber) which would go on a bog standard pentium III PC, which can go to low light levels? I don't need to 'see in the dark', just do dark field optical microscopy and some macro photography with the aperture closed right down to get a good depth of field.
Many thanks in advance
Richard ________________________________________ Richard Beanland Analytical Services Bookham Inc Caswell Towcester Northants NN12 8EQ United Kingdom Tel. +44 1327 356362 Fax. +44 1327 356775 http://www.bookham.com ________________________________________
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. =======================================================================
==============================Original Headers============================== 6, 31 -- From richard.beanland-at-bookham.com Tue Mar 21 07:58:23 2006 6, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 6, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2LDwMjn019156 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 07:58:23 -0600 6, 31 -- X-VirusChecked: Checked 6, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 6, 31 -- X-Msg-Ref: server-14.tower-78.messagelabs.com!1142949501!43371919!1 6, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 6, 31 -- X-Originating-IP: [213.249.209.179] 6, 31 -- Received: (qmail 5026 invoked from network); 21 Mar 2006 13:58:21 -0000 6, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 6, 31 -- by server-14.tower-78.messagelabs.com with SMTP; 21 Mar 2006 13:58:21 -0000 6, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 6, 31 -- Tue, 21 Mar 2006 13:58:20 +0000 6, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 31 -- Content-class: urn:content-classes:message 6, 31 -- MIME-Version: 1.0 6, 31 -- Content-Type: text/plain; 6, 31 -- charset="us-ascii" 6, 31 -- Subject: LM: integrating digital camera 6, 31 -- Date: Tue, 21 Mar 2006 13:58:19 -0000 6, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047CED-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 6, 31 -- X-MS-Has-Attach: 6, 31 -- X-MS-TNEF-Correlator: 6, 31 -- Thread-Topic: LM: integrating digital camera 6, 31 -- Thread-Index: AcZM74IXGTGLQD+wSZCV9dT4DMyAFw== 6, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 6, 31 -- To: {microscopy-at-microscopy.com} 6, 31 -- X-OriginalArrivalTime: 21 Mar 2006 13:58:20.0696 (UTC) FILETIME=[83189980:01C64CEF] 6, 31 -- Content-Transfer-Encoding: 8bit 6, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2LDwMjn019156 ==============================End of - Headers==============================
I am currently using one to observe electron diffraction. However, this camera only has VGA resolution (640 x 480 pixels). It has a USB interface, so you don't need a framegrabber. And the software it comes with is worth the $100 alone - you can use it to change the integration time of the CMOS sensor for low light levels.
I've been very happy with this little product, but you might look at Orion's other products if you want a higher-resolution camera.
Ben McMorran Research Associate, Atom Optics Group Department of Physics University of Arizona 1118 E 4th St Tucson, AZ 85721
ph. 520-621-2688
Quoting richard.beanland-at-bookham.com:
} ---------------------------------------------------------------------------- } 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 folks, } CCD camera technology has moved on a lot since I bought one for } our optical microscopes back in 1998. The camera still works fine, but } it has always struggled with low light levels (on-chip integration being } an expensive option in those days). Does anyone have any } recommendations for a relatively cheap camera (+ controller/software + } framegrabber) which would go on a bog standard pentium III PC, which can } go to low light levels? I don't need to 'see in the dark', just do dark } field optical microscopy and some macro photography with the aperture } closed right down to get a good depth of field. } } Many thanks in advance } } Richard } ________________________________________ } Richard Beanland } Analytical Services } Bookham Inc } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } } ======================================================================= } This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly prohibited. } No part of this message can be considered a request for goods or } services. } ======================================================================= } } } ==============================Original Headers============================== } 6, 31 -- From richard.beanland-at-bookham.com Tue Mar 21 07:58:23 2006 } 6, 31 -- Received: from mail78.messagelabs.com } (mail78.messagelabs.com [195.245.230.131]) } 6, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2LDwMjn019156 } 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 07:58:23 -0600 } 6, 31 -- X-VirusChecked: Checked } 6, 31 -- X-Env-Sender: richard.beanland-at-bookham.com } 6, 31 -- X-Msg-Ref: server-14.tower-78.messagelabs.com!1142949501!43371919!1 } 6, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- } 6, 31 -- X-Originating-IP: [213.249.209.179] } 6, 31 -- Received: (qmail 5026 invoked from network); 21 Mar 2006 } 13:58:21 -0000 } 6, 31 -- Received: from unknown (HELO } cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 6, 31 -- by server-14.tower-78.messagelabs.com with SMTP; 21 Mar } 2006 13:58:21 -0000 } 6, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI } ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with } Microsoft SMTPSVC(6.0.3790.211); } 6, 31 -- Tue, 21 Mar 2006 13:58:20 +0000 } 6, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 6, 31 -- Content-class: urn:content-classes:message } 6, 31 -- MIME-Version: 1.0 } 6, 31 -- Content-Type: text/plain; } 6, 31 -- charset="us-ascii" } 6, 31 -- Subject: LM: integrating digital camera } 6, 31 -- Date: Tue, 21 Mar 2006 13:58:19 -0000 } 6, 31 -- Message-ID: } {9645D3E33E4C6548B12A7B25F611533E047CED-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} } 6, 31 -- X-MS-Has-Attach: } 6, 31 -- X-MS-TNEF-Correlator: } 6, 31 -- Thread-Topic: LM: integrating digital camera } 6, 31 -- Thread-Index: AcZM74IXGTGLQD+wSZCV9dT4DMyAFw== } 6, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 6, 31 -- To: {microscopy-at-microscopy.com} } 6, 31 -- X-OriginalArrivalTime: 21 Mar 2006 13:58:20.0696 (UTC) } FILETIME=[83189980:01C64CEF] } 6, 31 -- Content-Transfer-Encoding: 8bit } 6, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k2LDwMjn019156 } ==============================End of - Headers==============================
==============================Original Headers============================== 13, 27 -- From mcmorran-at-physics.arizona.edu Tue Mar 21 09:19:56 2006 13, 27 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 13, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2LFJt0s029926 13, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 09:19:55 -0600 13, 27 -- Received: from localhost (gimli.email.arizona.edu [10.0.0.223]) 13, 27 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 564C0D415A7 13, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 08:19:55 -0700 (MST) 13, 27 -- Received: from localhost (treebeard.email.arizona.edu [10.0.0.213]) 13, 27 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9324BD50DCC 13, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 08:19:53 -0700 (MST) 13, 27 -- Received: from dhcp029.pas.arizona.edu (dhcp029.pas.arizona.edu 13, 27 -- [150.135.51.31]) by www.email.arizona.edu (Horde) with HTTP for 13, 27 -- {mcmorran-at-email.arizona.edu} ; Tue, 21 Mar 2006 08:19:53 -0700 13, 27 -- Message-ID: {20060321081953.2ssjggcg4g88s44o-at-www.email.arizona.edu} 13, 27 -- X-Priority: 3 (Normal) 13, 27 -- Date: Tue, 21 Mar 2006 08:19:53 -0700 13, 27 -- From: Ben McMorran {mcmorran-at-physics.arizona.edu} 13, 27 -- To: Microscopy-at-microscopy.com 13, 27 -- Subject: Re: [Microscopy] LM: integrating digital camera 13, 27 -- References: {200603211401.k2LE1kmh023763-at-ns.microscopy.com} 13, 27 -- In-Reply-To: {200603211401.k2LE1kmh023763-at-ns.microscopy.com} 13, 27 -- MIME-Version: 1.0 13, 27 -- Content-Type: text/plain; charset="ISO-8859-1"; format="flowed" 13, 27 -- Content-Disposition: inline 13, 27 -- Content-Transfer-Encoding: 7bit 13, 27 -- User-Agent: Internet Messaging Program (IMP) 4.0-cvs 13, 27 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Sputter coaters will often take targets made of different metals and this is what primarily determines fineness of the "grain", along with sputtering current, sputtering atmosphere, and coating times. We currently have targets made of platinum, chromium, gold, tantalum, titanium, and aluminum. For SEM conductive coating, we generally use platinum, which is much finer-grained than gold or gold/palladium. Finer yet is chromium, but that has the disadvantage of oxidizing rapidly to form an insulating layer, which almost makes it a "one-viewing" type of coating, unless the sample is stored under a good vacuum. Also, oxidizing metals require that the sputter coater be able to etch the oxide layer off the target before the actual coating cycle.
Iridium and osmium are also fine-grained coatings, but I have no personal experience with these metals. Osmium coating requires a special instrument, which is supplied by SPI Supplies and maybe others.
If you have multiple users, as we do, repeatability is important, but so is flexibility. We are constantly changing coating times and sputtering currents, depending on sample, desired magnification, and other factors. The ability to take targets of different metals is important----some require different sputtering parameters. Also, if your facility is like ours, word might get out to the electrical engineering folks that a coater is available to put down repeatable layers of metals on various substrates for making circuits and other "dark side" wizardry. If so, be prepared for some exotic requests.
Finally, if you do end up coating with a variety of metals, you can save tons of money by buying custom targets from 3rd party suppliers, such as Abe Dayani of Refining Systems, Inc. and possibly others. (No financial interest, etc., but just a satisfied customer.) Such suppliers can provide targets in almost any metal, in any size, and in any purity needed, at substantial savings over OEM prices.
Turbo coaters are definitely nice and I imagine that any coater in that price range would also provide the ability to play with coating parameters for most applications. I also recommend a rotating, tiltable specimen stage.
Hope some of this helps.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: nicol-at-semiconductor.com [mailto:nicol-at-semiconductor.com] Sent: Tuesday, March 21, 2006 7:33 AM To: Tindall, Randy D.
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Email: nicol-at-semiconductor.com Name: Nick Aitken
Title-Subject: [Filtered] sputter coaters
Question: Hi all,
we are currently looking at replacing our old Hummer X coater. It is not producing coatings that look good enough at high magnification. We are currently looking at a couple of turbo pumped coaters by various manufacturers, but I thought I would get some external opinions. First of all our requirements: We need a good conductive coating that will not show a lot of grain at around 250 thousand times mag : repeatability is a must. We have multiple users preparing many samples and we need consistent results across the board
What manufacturers should we be looking at, and what target material would be the best for this type of use? We would be coating semiconductor devices in both topographical and cross sectional orientations.
Cr gives one of the best coatings, if not the best, for high resolution. (I highly suspect that a well known fellow from SPI will probably disagree with that statement, again.) However, it oxidizes in short time and the sample should not be exposed to air for prolonged periods of time. We have introduced the SampleSaver(TM) Storage Container system that will solve that problem. The next target materials that people use with good success is Ir and W. These also give a fine grain size and are less susceptible to oxidation. It is important that your coating system gives a very thin, but uniform coating that conforms to the surface. This basically allows the charge to be drained from the surface. With these thin, high resolution coatings, it is also important for insulating materials to keep the accelerating voltage down so that the beam is not penetrating deep into the samples and causing the charge to be trapped well below the coating where it is unable to be dissipated to ground.
Since your application is with semiconductor materials, the ability to have an etch gun is a definite plus. With our system, the etch gun can be used to low angle polish as well as etch at higher angles. The higher angle will give a differential sputter etch that can enhance contrast between different materials as well as etch grain structures in your coatings.
Disclaimer: South Bay Technology, Inc. manufactures and sells the IBS/e system and the SampleSaver(TM) Storage Container.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: nicol-at-semiconductor.com [mailto:nicol-at-semiconductor.com] Sent: Tuesday, March 21, 2006 5:35 AM To: Walck-at-SouthBayTech.com
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Email: nicol-at-semiconductor.com Name: Nick Aitken
Title-Subject: [Filtered] sputter coaters
Question: Hi all,
we are currently looking at replacing our old Hummer X coater. It is not producing coatings that look good enough at high magnification. We are currently looking at a couple of turbo pumped coaters by various manufacturers, but I thought I would get some external opinions. First of all our requirements: We need a good conductive coating that will not show a lot of grain at around 250 thousand times mag : repeatability is a must. We have multiple users preparing many samples and we need consistent results across the board
What manufacturers should we be looking at, and what target material would be the best for this type of use? We would be coating semiconductor devices in both topographical and cross sectional orientations.
I lost my second Anatech Hummer II a couple years ago did a similar search for replacements. I wound up with a Denton Desk II. It worked very well for about a year and then failed to fire or keep a steady plasma. Denton replaced the HV board on a trial swap basis but it made no difference. So something else was going on. However, as I was moving to smaller node sizes, HC contamination was becoming a real nusance. So this moved me to search for a turbo coater for finer grain at high mag.
I wound up getting a Denton Desk IV TSC with tilt and rotation. My major modification that they accommodated was the replacement of the oil diaphram pump with an Edwards XDS5 dry scroll pump. So the forepump is external from the desk-top coater. Total prep time is longer since the scroll pump does not pump as fast as an oil pump. But the results are excellent. If the TSC circuitry would support it, an XDS10 would likely be faster. Since my SEM is totally dry pumped, the whole prep scenario results in as close to zero HC contamination as possible.
The TSC uses 6cm diameter targets. Or you could use 5.75cm. The material to be used is tricky. Au produces a web or mesh coating and is not at all good for high mag. Au/Pd is good as is Pd. Better noble metals such as Pt and Ir are even better. But here is the kicker. If you do EDS, suppose you coat with Pt and the barrier layer is Pt. If you do quant in the barrier layer, it will be wrong due to the additional coating. W plugs are also an issue since its L beta is close to L alpha of Pt. I suppose my point is to not use a target material that is likely to be part of the IC. The other consideration is peak pileup at low eV. I wound up choosing Ir.
Protocol is:
put specimen in coater and close lid start rough pump wait until vac is about 322mT start turbo after Turbo -at- Speed is indicated, check terminal vacuum. Should be about 4-5mT open gas (Ar) adjust needle valve for vacuum of 15mT-20mT (depends on what final results you want) go to timed sputter set Rotate=20% (if you want rotation) set Power=60% (typically results in 12-15mA) set Time=60 seconds (adjust for your application) when done, turn off turbo (HV will already have gone off) open needle valve wide open (this increases friction at the turbo bearings to slow it down) When you hear the turbo running down to very slow, stop fore pump wait a few seconds and open the lid
This process takes longer than with the oil fore pump as I said. But there is no HC contamination as-prepared. As the specimen is left outside of the SEM and not under vacuum, there will be contamination and you will get the HC scan "burns." If you use the specimen again, stick it under a high intensity UV lamp for about two hours and see if that clears the HC.
Au/Pd, Pt, Ir range in price from about $300 to $1,000, in that order. However, the Ir target is quite thick. The others are around .008". Thickness obviously affects the price as does material and size. Abe offers targets in several thicknesses. The targets are not 99.999% pure. Each target will have a different purity and its own set of trace elements. So far, the worst target purity is 99.5% for the Pd and Ir. Trace elements are Mn, Si, Cu, Fe, P, S. But a 100-200A film is not likely to show up with such small trace values. The 0.5% trace is divided up amongst several trace elements--not just one.
Disclaimer: I have no financial interest in Denton or Refining Systems other than to hope they stay around to offer good products and good service.
Happy coating, gary g.
At 05:34 AM 3/21/2006, you wrote:
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==============================Original Headers============================== 19, 20 -- From gary-at-gaugler.com Tue Mar 21 12:25:23 2006 19, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 19, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2LIPNM1029314 19, 20 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 12:25:23 -0600 19, 20 -- Received: (qmail 2136 invoked from network); 21 Mar 2006 10:25:22 -0800 19, 20 -- Received: by simscan 1.1.0 ppid: 2132, pid: 2133, t: 0.2250s 19, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 19, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 19, 20 -- by qsmtp3 with SMTP; 21 Mar 2006 10:25:22 -0800 19, 20 -- Message-Id: {6.2.3.4.2.20060321093615.023c3770-at-mail.calweb.com} 19, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 19, 20 -- Date: Tue, 21 Mar 2006 10:25:25 -0800 19, 20 -- To: nicol-at-semiconductor.com 19, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 19, 20 -- Subject: Re: [Microscopy] viaWWW: sputter coaters 19, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 19, 20 -- In-Reply-To: {200603211334.k2LDY6II012454-at-ns.microscopy.com} 19, 20 -- References: {200603211334.k2LDY6II012454-at-ns.microscopy.com} 19, 20 -- Mime-Version: 1.0 19, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
As a consultant I am often asked to appraise a laboratory and its staff. X-from this, with my interests in Quality in Electron Microscopy, I am involved with a paper related to the running of EM units and to training levels. The referees are happy with the paper but would like to see some results from a range of different units. They are interested in which are areas of action in a laboratory and which are the areas that do not seem to be active.
We have put together an EM Unit Survey on an Excel file where through the totting up of points we will be able to see where laboratories stand. Knowing how political these results could be I have arranged for a kart race colleague to collate them for us and to discarding their email source once the Excel file has been collected. In this way we will be able to obtain information but neither the authors of the paper nor others will know the source of the data. The Excel file is available on the Protrain web site under the Hints and Tips section for those who wish to help us.
If you would be able to help us it requires selecting your response to the Excel file and forwarding it to info-at-whiltonmillkartclub.co.uk .
Many thanks if you are able to help. If you would like to receive the data when collated please mail me separately.
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7802 966067 Web www.emcourses.com
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This Question was submitted to Ask-A-Microscopist by (jminarcik-at-sbcglobal.net) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, March 21, 2006 at 15:49:27 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both jminarcik-at-sbcglobal.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: jminarcik-at-sbcglobal.net Name: John Minarcik
Organization: Hard Knox
Education: Graduate College
Location: City, State, Country
Title: Plastic embedded super high quality H&E histo slides
Question: I need a set of super high quality plastic-embedded general H&E slides covering all types of tissues/organs.
We are looking for a disposable biopsy punch to use for cutting samples from flower petals and other plant material. I have found references to the Harris Uni-core punch and to the Miltex punches (used by our VET school).
I would appreciate hearing from anyone who has used either type (or has a recommendation for another type) and a source for ordering them. We would like to use them more than once if possible. I am sure number of uses depends on type of sample.
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============================== 6, 21 -- From dsherman-at-purdue.edu Tue Mar 21 20:43:40 2006 6, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 6, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M2hdZC011497 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 20:43:39 -0600 6, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Tue, 21 Mar 2006 21:43:38 -0500 6, 21 -- Received: from 12.222.30.139 ([12.222.30.139]) 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 ; 6, 21 -- Wed, 22 Mar 2006 02:33:55 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 6, 21 -- Date: Tue, 21 Mar 2006 21:33:54 -0500 6, 21 -- Subject: Biopsy punch 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C04621C2.12C7%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Biopsy punch 6, 21 -- Thread-Index: AcZNWQ/ZTi8fSblMEdqUkwAKlcoUxg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 22 Mar 2006 02:43:38.0969 (UTC) FILETIME=[6C850490:01C64D5A] ==============================End of - Headers==============================
I am very happy to report that the Facility Organization and Management FIG is sponsoring a special pre-meeting workshop in Chicago on Saturday July 29 from 1-5pm prior to the start of M&M 2006. Information is posted on the MSA web site under annual meeting-2006. Select Pre-meeting events.
http://mm2006.microscopy.org/
I am really excited about the workshop as it will cover topics quite different than those covered in past sessions on Core Facility Management. The presenters are all professionals from the business and management areas, not microscopists. Having a workshop run by non-microscopists but professionals in management can help us think ³out of the box² as we develop new strategies for helping keep our facilities viable.
Facility Operations and Management FIG Workshop
Saturday, July 29, 1:00-5:00pm (prior to M&M 2006) Holiday Inn City Center Hotel, Chicago, IL Registration fee: $40 Space is limited. You do not have to be a FOM FIG member to attend the workshop. However, you must download the form from the MSA meeting website and submit it with your registration fee.
The Workshop title is: New Approaches to Marketing, Managing, and Money for Maintaining a Core Facility (4M¹s)
Topics: 1) How to Make a Business Plan for short and Long-term Facility Maintenance and Growth. Donald Blewett, Associate Director, Burton Morgan Center for Entrepreneurship, Purdue University
3) Marketing a Facility to Increase and Maintain a User Base (and maintain the support of the upper administration). Dr. George Adams, Research Development manager, Birck nanotechnology Center, Purdue University
4) Developing a financial plan for the long-term ³care and feeding² of Major Equipment. Charlene Sullivan, Professor, Krannert School of Management, Purdue University Note: extensive research and information from many microscopy facilities have provided the raw data for the analysis needed to develop this plan. The results will be presented at the workshop.
Hope to see many of you there.
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://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 16, 22 -- From dsherman-at-purdue.edu Tue Mar 21 21:04:59 2006 16, 22 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 16, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M34w3v015807 16, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 21:04:58 -0600 16, 22 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 16, 22 -- Tue, 21 Mar 2006 22:04:58 -0500 16, 22 -- Received: from 12.222.30.139 ([12.222.30.139]) 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 ; 16, 22 -- Wed, 22 Mar 2006 03:04:58 +0000 16, 22 -- User-Agent: Microsoft-Entourage/11.2.1.051004 16, 22 -- Date: Tue, 21 Mar 2006 22:04:57 -0500 16, 22 -- Subject: Facility Management Workshop-M&M2006 16, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 16, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 16, 22 -- Message-ID: {C0462909.12D2%dsherman-at-purdue.edu} 16, 22 -- Thread-Topic: Facility Management Workshop-M&M2006 16, 22 -- Thread-Index: AcZNXWZJpKSgCrlQEdqUkwAKlcoUxg== 16, 22 -- Mime-version: 1.0 16, 22 -- Content-type: text/plain; 16, 22 -- charset="ISO-8859-1" 16, 22 -- X-OriginalArrivalTime: 22 Mar 2006 03:04:58.0438 (UTC) FILETIME=[67247E60:01C64D5D] 16, 22 -- Content-Transfer-Encoding: 8bit 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2M34w3v015807 ==============================End of - Headers==============================
Well, I am probably not the guy who Scott referred to but I think that one can't really talk sensibly about coating for the SEM without talking about beam kV. As some of you may remember, when the first immersion-lens FE scopes arrived, I was one of the first to champion 1.5 kV for HR SEM of biological specimens. At low kV, you don't need much conductivity because the SE coef if higher and you don't need so much beam current because the contrast is higher to, but you do still usually need some coating (for surface contrast, even if nothing else.).
We may have made our Cr coats wrong but David Joy once opined that one can't have a "few-nw-thick" film of Cr and he looked at a lot of them in in TEM with energy loss to detect the oxide. Cr will oxidize in a flash, even in microscope vacuum (remember, 10*-6 torr is one monolayer/second and a lot of that monolayer is likely to be water vapor, which becomes oxygen if the beam hits it. You may have a cold trap but, as the "ice" that forms on it is amorphous, it has a much higher vapor pressure than you think.)
So we could never get Cr to work. Specimens charged and the contrast was not great. Also, it is important to remember that, as Cr-oxide is about 3x less dense than Cr metal, the geometric thickness is about 3x thicker than that indicated by the crystal monitor on your coater.
We could not see shall surface structures.
Instead, we used ion-beam sputtered Pt. It does form 1-2 nm crystals (about 3-5 atoms on a side) about 1-2 nm apart when put on at about 1 mn average thickness. You see this pattern when you look at the specimen at 5kV where the beam is smaller and Z-contrast makes them visible, even in SE, but the surface contrast is rudimentary.
Fortunately, these crystals are usually not visible at 1.5kV and they certainly keep the charging under control and provide some surface contrast. We all like "resolution" but remember that few specimen preparation methods preserve structure below 3nm anyway.
And Pt films are MUCH less reactive than Cr.
Not to push it but metals conduct BECAUSE of their crystalline structure. Amorphous metals are much less conductive. Metal oxides less still. Cr films have a conductivity similar to that of carbon films.
Many people use Cr at high kV but I will leave it to others to discuss the pro's and con's.
Cheers,
Jim Pawley (not only a confocalite!)
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-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 10-22, 2006, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2006 "If it ain't diffraction, it must be statistics." Anon.
==============================Original Headers============================== 15, 27 -- From jbpawley-at-wisc.edu Tue Mar 21 23:50:00 2006 15, 27 -- Received: from smtp5.wiscmail.wisc.edu (heimdall.doit.wisc.edu [144.92.197.159]) 15, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M5nxK1001783 15, 27 -- for {Microscopy-at-Microscopy.Com} ; Tue, 21 Mar 2006 23:49:59 -0600 15, 27 -- Received: from avs-daemon.smtp5.wiscmail.wisc.edu by smtp5.wiscmail.wisc.edu 15, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.08 (built Sep 22 2005)) 15, 27 -- id {0IWI00D17LJBAZ-at-smtp5.wiscmail.wisc.edu} for Microscopy-at-Microscopy.Com; 15, 27 -- Tue, 21 Mar 2006 23:49:59 -0600 (CST) 15, 27 -- Received: from [172.16.1.41] ([144.92.238.207]) by smtp5.wiscmail.wisc.edu 15, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.08 (built Sep 22 2005)) 15, 27 -- with ESMTPSA id {0IWI00K1XLJ87C-at-smtp5.wiscmail.wisc.edu} ; Tue, 15, 27 -- 21 Mar 2006 23:49:57 -0600 (CST) 15, 27 -- Date: Tue, 21 Mar 2006 23:49:52 -0600 15, 27 -- From: James Pawley {jbpawley-at-wisc.edu} 15, 27 -- Subject: [Microscopy] RE: viaWWW: sputter coaters 15, 27 -- In-reply-to: {200603211721.k2LHLHHZ027169-at-ns.microscopy.com} 15, 27 -- X-Sender: jbpawley-at-wiscmail.wisc.edu 15, 27 -- To: walck-at-southbaytech.com, 15, 27 -- "ListServer-at-MSA.Microscopy.Com" {Microscopy-at-Microscopy.Com} 15, 27 -- Message-id: {p06110417c0468f14149e-at-[172.16.1.41]} 15, 27 -- MIME-version: 1.0 15, 27 -- Content-type: text/plain; format=flowed; charset=us-ascii 15, 27 -- Content-transfer-encoding: 7BIT 15, 27 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=[144.92.238.207] 15, 27 -- X-Spam-PmxInfo: Server=avs-1, Version=5.1.2.240295, Antispam-Engine: 2.3.0.1, 15, 27 -- Antispam-Data: 2006.3.21.213107, SenderIP=[144.92.238.207] 15, 27 -- References: {200603211721.k2LHLHHZ027169-at-ns.microscopy.com} ==============================End of - Headers==============================
Please note... the speaker topics were miss numbered (thank WORD for auto correcting again!). There are only 3 speakers. Sorry for clogging your in box twice with the same message.
Debby
} From: {dsherman-at-purdue.edu} } Reply-To: {dsherman-at-purdue.edu} } Date: Tue, 21 Mar 2006 21:38:33 -0600 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] Facility Management Workshop-M&M2006 } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } List members, } } I am very happy to report that the Facility Organization and Management FIG } is sponsoring a special pre-meeting workshop in Chicago on Saturday July 29 } from 1-5pm prior to the start of M&M 2006. Information is posted on the } MSA web site under annual meeting-2006. Select Pre-meeting events. } } http://mm2006.microscopy.org/ } } I am really excited about the workshop as it will cover topics quite } different than those covered in past sessions on Core Facility Management. } The presenters are all professionals from the business and management areas, } not microscopists. Having a workshop run by non-microscopists but } professionals in management can help us think ³out of the box² as we develop } new strategies for helping keep our facilities viable. } } } Facility Operations and Management FIG Workshop } } Saturday, July 29, 1:00-5:00pm (prior to M&M 2006) } Holiday Inn City Center Hotel, Chicago, IL } Registration fee: $40 Space is limited. } You do not have to be a FOM FIG member to attend the workshop. However, you } must download the form from the MSA meeting website and submit it with your } registration fee. } } } The Workshop title is: } New Approaches to Marketing, Managing, and Money for Maintaining a } Core Facility (4M¹s) } } Topics: } 1) How to Make a Business Plan for short and Long-term Facility } Maintenance and Growth. } Donald Blewett, Associate Director, Burton Morgan Center for } Entrepreneurship, Purdue University } } 2) Marketing a Facility to Increase and Maintain a User Base (and } maintain the support of the upper administration). } Dr. George Adams, Research Development manager, Birck nanotechnology Center, } Purdue University } } 3) Developing a financial plan for the long-term ³care and feeding² of } Major Equipment. } Charlene Sullivan, Professor, Krannert School of Management, Purdue } University } Note: extensive research and information from many microscopy facilities } have provided the raw data for the analysis needed to develop this plan. The } results will be presented at the workshop. } } Hope to see many of you there. } } 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://www.agriculture.purdue.edu/microscopy } } } } ==============================Original Headers============================== } 16, 22 -- From dsherman-at-purdue.edu Tue Mar 21 21:04:59 2006 } 16, 22 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu } [128.210.63.223]) } 16, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M34w3v015807 } 16, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 21:04:58 -0600 } 16, 22 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by } exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 16, 22 -- Tue, 21 Mar 2006 22:04:58 -0500 } 16, 22 -- Received: from 12.222.30.139 ([12.222.30.139]) 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 ; } 16, 22 -- Wed, 22 Mar 2006 03:04:58 +0000 } 16, 22 -- User-Agent: Microsoft-Entourage/11.2.1.051004 } 16, 22 -- Date: Tue, 21 Mar 2006 22:04:57 -0500 } 16, 22 -- Subject: Facility Management Workshop-M&M2006 } 16, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} } 16, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 16, 22 -- Message-ID: {C0462909.12D2%dsherman-at-purdue.edu} } 16, 22 -- Thread-Topic: Facility Management Workshop-M&M2006 } 16, 22 -- Thread-Index: AcZNXWZJpKSgCrlQEdqUkwAKlcoUxg== } 16, 22 -- Mime-version: 1.0 } 16, 22 -- Content-type: text/plain; } 16, 22 -- charset="ISO-8859-1" } 16, 22 -- X-OriginalArrivalTime: 22 Mar 2006 03:04:58.0438 (UTC) } FILETIME=[67247E60:01C64D5D] } 16, 22 -- Content-Transfer-Encoding: 8bit } 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k2M34w3v015807 } ==============================End of - Headers==============================
==============================Original Headers============================== 6, 23 -- From dsherman-at-purdue.edu Wed Mar 22 08:19:52 2006 6, 23 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2MEJp1q027971 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 08:19:51 -0600 6, 23 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 23 -- Wed, 22 Mar 2006 09:19:51 -0500 6, 23 -- 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 ; 6, 23 -- Wed, 22 Mar 2006 14:19:51 +0000 6, 23 -- User-Agent: Microsoft-Entourage/11.2.1.051004 6, 23 -- Date: Wed, 22 Mar 2006 09:19:51 -0500 6, 23 -- Subject: Correction- Facility Management Workshop-M&M2006 6, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 23 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 23 -- Message-ID: {C046C737.E4EE%dsherman-at-purdue.edu} 6, 23 -- Thread-Topic: Correction- Facility Management Workshop-M&M2006 6, 23 -- Thread-Index: AcZNu66X7Q7MormuEdq0UwARJN08Mg== 6, 23 -- In-Reply-To: {200603220338.k2M3cXN7024849-at-ns.microscopy.com} 6, 23 -- Mime-version: 1.0 6, 23 -- Content-type: text/plain; 6, 23 -- charset="ISO-8859-1" 6, 23 -- X-OriginalArrivalTime: 22 Mar 2006 14:19:51.0360 (UTC) FILETIME=[AECE0C00:01C64DBB] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2MEJp1q027971 ==============================End of - Headers==============================
Are there tight junctions (zonula occludens) in keratinized stratified squamous epithelia such as skin? Or do the lamellar granules (membrane coating granules) take care of all the sealing needed? As a corollary- are there tight junctions in non-keratinizing stratified epithelia such as esophagus? thanks for some basic histo I should know! tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have tried to give away a bunch of photo paper we don't use any more with no luck.
It is a little old, a couple of years, but it has been in a refrigerator and should be OK. It's not getting any younger and I don't think we are ever going to use it.
It is Kodak Kodabrome II RC in various contrast grades, 1 -5.
If you can use it, contact me and we can work out some way to get it to you.
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============================== 7, 21 -- From jmkrupp-at-ucsc.edu Wed Mar 22 12:01:36 2006 7, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2MI1XTt010362 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 12:01:33 -0600 7, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 7, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.1/8.13.1) with ESMTP id k2MHlobo017450 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 09:47:51 -0800 (PST) 7, 21 -- Received: from [128.114.25.214] (account jmkrupp-at-ucsc.edu [128.114.25.214] verified) 7, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 7, 21 -- with ESMTPA id 50032116 for microscopy-at-microscopy.com; Wed, 22 Mar 2006 09:47:50 -0800 7, 21 -- Mime-Version: 1.0 7, 21 -- Message-Id: {p06230902c0473d8062cf-at-[128.114.25.214]} 7, 21 -- Date: Wed, 22 Mar 2006 09:47:49 -0800 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 7, 21 -- Subject: Surplus photo paper 7, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 7, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 7, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 7, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (alvarobq-at-fcien.edu.uy) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, March 22, 2006 at 11:08:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both alvarobq-at-fcien.edu.uy as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I`ll very appreciate if you can send me the e-mail address of a Quetol 651 supplier. I need to know more about this resin to compare to Spurr. If every one have some experience, please send me any advise. Thank you very much.
This Question was submitted to Ask-A-Microscopist by (alvarobq-at-fcien.edu.uy) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, March 22, 2006 at 11:08:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both alvarobq-at-fcien.edu.uy as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I`ll very appreciate if you can send me the e-mail address of a Quetol 651 supplier. I need to know more about this resin to compare to Spurr. If every one have some experience, please send me any advise. Thank you very much.
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 rpetrova-at-mail.ucf.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpetrova-at-mail.ucf.edu Name: Rumy
Organization: UCF
Title-Subject: [Filtered] sample prep
Question: Does anyone know what is the best way to polish CdZnO ? Would the wedge polishing work,using diamond lapping film?
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 v_bleu_knight-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Knives for Ultrathin sectioning of bone or calcium crystals
Question: Hello List,
Does anyone have experience creating ultrathin sections of tissues that have bone or calcium crystals in them? If so, what kind of diamond knife are you using? I am reluctant to use my standard 45 degree diamond knife because I feel that it would dull very quickly. Thank you in advance for your response.
Sincerely, Bleu Knight PhD Candidate New Mexico State University
I have tried to give away a bunch of photo paper we don't use any more with no luck.
It is a little old, a couple of years, but it has been in a refrigerator and should be OK. It's not getting any younger and I don't think we are ever going to use it.
It is Kodak Kodabrome II RC in various contrast grades, 1 -5.
If you can use it, contact me and we can work out some way to get it to you.
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============================== 7, 21 -- From jmkrupp-at-ucsc.edu Wed Mar 22 18:39:50 2006 7, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2MI1XTt010362 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 12:01:33 -0600 7, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 7, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.1/8.13.1) with ESMTP id k2MHlobo017450 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 09:47:51 -0800 (PST) 7, 21 -- Received: from [128.114.25.214] (account jmkrupp-at-ucsc.edu [128.114.25.214] verified) 7, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 7, 21 -- with ESMTPA id 50032116 for microscopy-at-microscopy.com; Wed, 22 Mar 2006 09:47:50 -0800 7, 21 -- Mime-Version: 1.0 7, 21 -- Message-Id: {p06230902c0473d8062cf-at-[128.114.25.214]} 7, 21 -- Date: Wed, 22 Mar 2006 09:47:49 -0800 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 7, 21 -- Subject: Surplus photo paper 7, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 7, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 7, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 7, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
Are there tight junctions (zonula occludens) in keratinized stratified squamous epithelia such as skin? Or do the lamellar granules (membrane coating granules) take care of all the sealing needed? As a corollary- are there tight junctions in non-keratinizing stratified epithelia such as esophagus? thanks for some basic histo I should know! tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I'm looking into writing javascript for reading this SEM data (e.g., magnification) that is embedded in the TIFF files in a non-standard way. I expect the javascript to work within Photoshop, and I'd certainly be interested if anyone has already done this and can provide an example (although I am sure my SEM data is formatted differently). (BTW, I already aware I can open these files with a text editor and simply extract the text ... But that's not very elegant ...)
The 2nd issue is what to do with the data once retrieved? I could simply write it to a text file, but it would be better to write it back to the TIFF in a standard way. For example, to put it all in the EXIF "comments" field ... Or even better, to put it where it should be put ... Except I can find no information as to Microscopists, as a group, asking that standard EXIF fields be designated and standardized (e.g., the "microns per pixel" field).
When we upgraded the computer and software for our JEOL 5600, I was so disappointed that the information that can be embedded into images was still in the 1970s style, taking up a significant portion of the bottom of the image, and not adjustable in any way, that I bit the bullet and wrote my own program to add a strip at the bottom of the images with information from the SEM data text file. This extended the dimensions of the 1280x960 image to 1280x1024, but none of the image area is taken up by those blocky looking, primitive characters. You can see examples from the page I recently posted from the "mystery object/starch grain" thread a month or so back:
http://www.mta.ca/dmf/download/ehrman/mystery.htm
In writing this, I was also wondering how "standard" some of this information is across instrument models and manufacturers? If people are interested, send me an email with one of your text files, along with the instrument make and model, I can do some comparisons. Along those lines, would anybody be interested in an application that would write this information to their images? If their is some consistency in the formats, it shouldn't take much to modify what I have already.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
michael-at-shaffer.net 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 use the term "metadata" loosely ... } } I'm looking into writing javascript for reading this SEM data (e.g., } magnification) that is embedded in the TIFF files in a non-standard way. I } expect the javascript to work within Photoshop, and I'd certainly be } interested if anyone has already done this and can provide an example } (although I am sure my SEM data is formatted differently). (BTW, I already } aware I can open these files with a text editor and simply extract the text } ... But that's not very elegant ...) } } The 2nd issue is what to do with the data once retrieved? I could simply } write it to a text file, but it would be better to write it back to the TIFF } in a standard way. For example, to put it all in the EXIF "comments" field } ... Or even better, to put it where it should be put ... Except I can find } no information as to Microscopists, as a group, asking that standard EXIF } fields be designated and standardized (e.g., the "microns per pixel" field). } } TIA :o) } } Cheerios, Michael Shaffer :o) } } SEM/MLA Research Coordinator } (709) 737-6799 (ofc) } (709) 737-6790 (lab) } (709) 737-6193 (FAX) } http://www.mun.ca/creait/maf/ } http://www.esd.mun.ca/epma/ } } Inco Innovation Centre } c/o Memorial University } 230 Elizabeth Avenue } P.O. Box 4200 } St. John's, NL A1C 5S7 } } } } } ==============================Original Headers============================== } 10, 19 -- From michael-at-shaffer.net Thu Mar 23 05:18:14 2006 } 10, 19 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) } 10, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NBIEjX025905 } 10, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 05:18:14 -0600 } 10, 19 -- Received: (qmail 6069 invoked from network); 23 Mar 2006 11:21:52 -0000 } 10, 19 -- Received: from unknown (HELO rarewolf1) (michael-at-shaffer.net-at-205.251.84.119) } 10, 19 -- by ws6-4.us4.outblaze.com with SMTP; 23 Mar 2006 11:21:52 -0000 } 10, 19 -- From: "michael shaffer" {michael-at-shaffer.net} } 10, 19 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} } 10, 19 -- Subject: SEM digital image "metadata" } 10, 19 -- Date: Thu, 23 Mar 2006 07:47:30 -0330 } 10, 19 -- Message-ID: {000d01c64e6b$612304f0$4f01a8c0-at-rarewolf1} } 10, 19 -- MIME-Version: 1.0 } 10, 19 -- Content-Type: text/plain; } 10, 19 -- charset="US-ASCII" } 10, 19 -- Content-Transfer-Encoding: 7bit } 10, 19 -- X-Mailer: Microsoft Office Outlook 11 } 10, 19 -- Thread-Index: AcZOa1+XpgOBO7+sRu65nUAmDJ856w== } 10, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 13, 19 -- From jehrman-at-mta.ca Thu Mar 23 07:27:46 2006 13, 19 -- Received: from mailserv.mta.ca (mailserv.mta.ca [138.73.1.1]) 13, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NDRkOX004970 13, 19 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 23 Mar 2006 07:27:46 -0600 13, 19 -- Received: from host-22-245.mta.ca ([138.73.22.245]) 13, 19 -- by mailserv.mta.ca with esmtp (Exim 4.52) 13, 19 -- id 1FMPqm-00039t-MN; Thu, 23 Mar 2006 09:27:40 -0400 13, 19 -- Message-ID: {4422A245.8090801-at-mta.ca} 13, 19 -- Date: Thu, 23 Mar 2006 09:27:33 -0400 13, 19 -- From: "James M. Ehrman" {jehrman-at-mta.ca} 13, 19 -- Organization: Mount Allison University 13, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 13, 19 -- MIME-Version: 1.0 13, 19 -- To: michael-at-shaffer.net, Microscopy Listserv {Microscopy-at-MSA.Microscopy.com} 13, 19 -- Subject: Re: [Microscopy] SEM digital image "metadata" 13, 19 -- References: {200603231118.k2NBItM9026890-at-ns.microscopy.com} 13, 19 -- In-Reply-To: {200603231118.k2NBItM9026890-at-ns.microscopy.com} 13, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Bleu Knight wrote: ================================================================== Does anyone have experience creating ultrathin sections of tissues that have bone or calcium crystals in them? If so, what kind of diamond knife are you using? I am reluctant to use my standard 45 degree diamond knife because I feel that it would dull very quickly. Thank you in advance for your response. ================================================================== So far as I know, all suppliers of diamond knives offer a version called a "materials science" diamond knife. However, some offer a product that has a higher knife angle (e.g. 55 instead of 45 deg.) but others, e.g. SPI offer the same angle (e.g. 45 deg) but with the caveat that the last of the "fine striations" have not been removed. We have never found these fine striations problematic since with the first pass of the knife over the sample, striations larger than these fine ones will be put into the knife edge anyhow. And since the final polishing step to take out the last of the fine striations is the most expensive, the cost of the SPI Supplies materials science knife is cheaper than that of a so-called "life science" knife.
We have found that the lower 45 deg angle results in sections easier to cut and with far fewer problems displaying "compression" effects.
You are correct in being reluctant to use your "standard" 45 deg knife because these kinds of samples will quite quickly put in striations that will render the knife useless for your other work.
You can find out more information about the SPI materials science diamond knife on URL http://www.2spi.com/catalog/knives/materials.shtml
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============================== 15, 24 -- From cgarber-at-2spi.com Thu Mar 23 08:18:01 2006 15, 24 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 15, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NEI1Cs015158 15, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Mar 2006 08:18:01 -0600 15, 24 -- Received: from ibm1x23g2abfyg ([210.22.189.66]) 15, 24 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k2NEHlPI030752 15, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Mar 2006 09:17:58 -0500 15, 24 -- X-IDV-FirstRcvd: [210.22.189.66] 15, 24 -- X-IDV-HELO: ibm1x23g2abfyg 15, 24 -- Message-ID: {012c01c64e84$9114d8f0$9f0aa8c0-at-ibm1x23g2abfyg} 15, 24 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 15, 24 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 15, 24 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 15, 24 -- Subject: Diamond knife for tissue with bone or calcium crystals 15, 24 -- Date: Thu, 23 Mar 2006 09:17:42 -0500 15, 24 -- MIME-Version: 1.0 15, 24 -- Content-Type: text/plain; 15, 24 -- charset="Windows-1252" 15, 24 -- X-Priority: 3 15, 24 -- X-MSMail-Priority: Normal 15, 24 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 15, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 15, 24 -- Content-Transfer-Encoding: 8bit 15, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2NEI1Cs015158 ==============================End of - Headers==============================
Thomas The answer is yes, tight junctions are a characteristic of vertebrate epithelia and as far as I know occur in all types whether keratinizing or not. Check your library for a copy of Porter & Bonneville's Fine Structure of Cells and Tissues for excellent EMs. Richard Harris Laboratory Supervisor, Imaging and Analysis Department of Biology, University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- X-from: phillipst-at-missouri.edu [mailto:phillipst-at-missouri.edu] Sent: Wednesday, March 22, 2006 7:47 PM To: rjharris-at-uwo.ca
Are there tight junctions (zonula occludens) in keratinized stratified squamous epithelia such as skin? Or do the lamellar granules (membrane coating granules) take care of all the sealing needed? As a corollary- are there tight junctions in non-keratinizing stratified epithelia such as esophagus? thanks for some basic histo I should know! tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
You may find that the magnification data is stored in your TIFF images in the XResolution and/or YResolution data which is a part of the standard TIFF structure. This data is stored as two long integers, the first representing the numerator, the second the denominator of a fractional number. There is another field called ResolutionUnit which gives the final result. The location of these fields within the TIFF file is found in directories in the file header. Noran used this method to store pixel size information in their Voyager/Vantage TIFF images. The full TIFF file specification can be found at http://partners.adobe.com/public/developer/en/tiff/TIFF6.pdf.
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==============================Original Headers============================== 6, 25 -- From djv23-at-cam.ac.uk Thu Mar 23 09:38:06 2006 6, 25 -- Received: from ppsw-1.csi.cam.ac.uk (ppsw-1.csi.cam.ac.uk [131.111.8.131]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NFc5vL002990 6, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 09:38:05 -0600 6, 25 -- X-Cam-SpamDetails: Not scanned 6, 25 -- X-Cam-AntiVirus: No virus found 6, 25 -- X-Cam-ScannerInfo: http://www.cam.ac.uk/cs/email/scanner/ 6, 25 -- Received: from focus.msm.cam.ac.uk ([131.111.100.73]:38831 helo=msm.cam.ac.uk) 6, 25 -- by ppsw-1.csi.cam.ac.uk (ppsw.cam.ac.uk [131.111.8.131]:25) 6, 25 -- with esmtp id 1FMRsm-0003cP-4K (Exim 4.54) 6, 25 -- (return-path {djv23-at-cam.ac.uk} ); Thu, 23 Mar 2006 15:37:52 +0000 6, 25 -- Received: from davids-pc.cam.ac.uk (sem-djv.msm.cam.ac.uk [131.111.100.208]) 6, 25 -- by msm.cam.ac.uk (8.13.6/8.13.6) with ESMTP id k2NFbnMR005393; 6, 25 -- Thu, 23 Mar 2006 15:37:52 GMT 6, 25 -- Message-Id: {6.2.1.2.0.20060323145258.02768bc0-at-focus.msm.cam.ac.uk} 6, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 6, 25 -- Date: Thu, 23 Mar 2006 15:40:38 +0000 6, 25 -- To: michael-at-shaffer.net 6, 25 -- From: David Vowles {djv23-at-cam.ac.uk} 6, 25 -- Subject: Re: [Microscopy] SEM digital image "metadata" 6, 25 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 6, 25 -- In-Reply-To: {200603231122.k2NBMF4o031271-at-ns.microscopy.com} 6, 25 -- References: {200603231122.k2NBMF4o031271-at-ns.microscopy.com} 6, 25 -- Mime-Version: 1.0 6, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Thanks to those with helpful comments about my question about whether there are tight junctions in either keratinized stratified squamous epithelia such as skin or non-keratinized stratified squamous epithelium such as esophagus. It is, as I expected, a controversial area. A tip led me to a couple of references from the Franke lab:
Langbein L. Grund C. Kuhn C. Praetzel S. Kartenbeck J. Brandner JM. Moll I. Franke WW. Tight junctions and compositionally related junctional structures in mammalian stratified epithelia and cell cultures derived therefrom. European Journal of Cell Biology. 81(8):419-35.
Schluter H. Wepf R. Moll I. Franke WW (2004) Sealing the live part of the skin: the integrated meshwork of desmosomes, tight junctions and curvilinear ridge structures in the cells of the uppermost granular layer of the human epidermis. European Journal of Cell Biology. 83(11-12):655-65.
I haven't gotten the full references yet but the abstracts state they "found an unexpected diversity of TJ-related structures" some of which show "colocalization with the most restricted transmembrane TJ marker protein, occludin,..." They report "TJ-related junctions are abundant..." is some stratified epithelia and that "most of them we have noticed, in addition, junctional regions showing relatively broad, ribbon-like membrane contacts which in cross-section often appear pentalaminar, with an electron-dense middle lamella ("lamellated TJs", coniunctiones laminosae)." Note that the abstract call these "TJ related junctions" and (since I haven't yet read the paper) implies to me that they are not necessarily classical TJ since they also note some of these are "characterized by a 10-30-nm dense lamina interposed between the two membranes" which is not something you would expect to see in a TJ. Thanks again for those who gave me tips to these and other references. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I am also interested to learn what methods are used in common brands of SEM to encode the scale or magnification information. We need this for our calibration and measurement software.
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 Headers============================== 3, 21 -- From donc-at-asmicro.com Thu Mar 23 11:04:34 2006 3, 21 -- Received: from smtp109.sbc.mail.re2.yahoo.com (smtp109.sbc.mail.re2.yahoo.com [68.142.229.96]) 3, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NH4XFi023944 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:04:33 -0600 3, 21 -- Received: (qmail 98485 invoked from network); 23 Mar 2006 17:04:31 -0000 3, 21 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.251.105.88 with login) 3, 21 -- by smtp109.sbc.mail.re2.yahoo.com with SMTP; 23 Mar 2006 17:04:30 -0000 3, 21 -- Message-ID: {015601c64e9b$63af7b60$c901a8c0-at-ASM11} 3, 21 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- To: "Microscopy List" {microscopy-at-microscopy.com} 3, 21 -- Subject: SEM digital image "metadata" 3, 21 -- Date: Thu, 23 Mar 2006 12:00:58 -0500 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: text/plain; 3, 21 -- charset="iso-8859-1" 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Priority: 3 3, 21 -- X-MSMail-Priority: Normal 3, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 3, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
} You may find that the magnification data is stored in your } TIFF images in the XResolution and/or YResolution data which } is a part of the standard TIFF structure.
Thanx for your response David! For clarification, when you say "standard TIFF structure", are you claiming that if Photoshop opens the TIFF, and if you save it as a different file, the information is still there? The TIFF definition allows for many variations. Whether TIFF reads & writes recognize these variations is another question.
Inco Innovation Centre c/o Memorial University 230 Elizabeth Avenue P.O. Box 4200 St. John's, NL A1C 5S7
} } ------------------------------------------------------------- } ---------- } } ----- 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 use the term "metadata" loosely ... } } } } I'm looking into writing javascript for reading this SEM data (e.g., } } magnification) that is embedded in the TIFF files in a non-standard } } way. I expect the javascript to work within Photoshop, and I'd } } certainly be interested if anyone has already done this and } can provide } } an example (although I am sure my SEM data is formatted } differently). } } (BTW, I already aware I can open these files with a text editor and } } simply extract the text ... But that's not very elegant ...) } } } } The 2nd issue is what to do with the data once retrieved? I could } } simply write it to a text file, but it would be better to } write it back } } to the TIFF in a standard way. For example, to put it all } in the EXIF } } "comments" field ... Or even better, to put it where it } should be put } } ... Except I can find no information as to Microscopists, as } a group, } } asking that standard EXIF fields be designated and } standardized (e.g., the "microns per pixel" field). } } } } TIA :o) } } } } Cheerios, Michael Shaffer :o) }
==============================Original Headers============================== 10, 21 -- From Michael-at-Shaffer.net Thu Mar 23 11:11:58 2006 10, 21 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) 10, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NHBucs001070 10, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:11:57 -0600 10, 21 -- Received: (qmail 11789 invoked from network); 23 Mar 2006 17:15:41 -0000 10, 21 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 10, 21 -- by ws6-4.us4.outblaze.com with SMTP; 23 Mar 2006 17:15:41 -0000 10, 21 -- From: "michael shaffer" {michael-at-Shaffer.net} 10, 21 -- To: "'David Vowles'" {djv23-at-cam.ac.uk} 10, 21 -- Cc: "'MSA Microscopy list'" {Microscopy-at-microscopy.com} 10, 21 -- Subject: RE: [Microscopy] SEM digital image "metadata" 10, 21 -- Date: Thu, 23 Mar 2006 13:41:53 -0330 10, 21 -- Message-ID: {001e01c64e9c$e3150c70$8d829986-at-roamingwolf} 10, 21 -- MIME-Version: 1.0 10, 21 -- Content-Type: text/plain; 10, 21 -- charset="us-ascii" 10, 21 -- Content-Transfer-Encoding: 7bit 10, 21 -- X-Mailer: Microsoft Office Outlook 11 10, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 10, 21 -- thread-index: AcZOkExcOc847R7uRlefsxlIg0+GAgAC9nuA 10, 21 -- In-Reply-To: {6.2.1.2.0.20060323145258.02768bc0-at-focus.msm.cam.ac.uk} ==============================End of - Headers==============================
Cerium Labs LLC., a wholly owned subsidiary of Spansion Inc. (NASDAQ: SPSN, former Flash Memory groups of AMD and Fujitsu,) has an immediate opening for a TEM analyst. Lab operates 2 DB-FIBs and 2 TEMs (JEM-2010 and CM300FEG+GIF) and a wide range of analytical equipment. Emphasis is being placed on experience: analytical capabilities, EDS, GIF-EELS, and documented hands-on operation of TEM in various modes including electron crystallography. Lab is providing services to a range of clients from semiconductor, alternative energy, and advanced-materials sectors.
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==============================Original Headers============================== 13, 31 -- From Jerzy.Gazda-at-ceriumlabs.com Thu Mar 23 11:34:06 2006 13, 31 -- Received: from amdext4.amd.com (amdext4.amd.com [163.181.251.6]) 13, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NHY5nv010885 13, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:34:06 -0600 13, 31 -- Received: from SAUSGW02.amd.com (sausgw02.amd.com [163.181.250.22]) 13, 31 -- by amdext4.amd.com (8.12.11/8.12.11/AMD) with ESMTP id k2NHY6jC015807 13, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:34:06 -0600 13, 31 -- Received: from 163.181.22.101 by SAUSGW01.amd.com with ESMTP (AMD SMTP 13, 31 -- Relay (Email Firewall v6.1.0)); Thu, 23 Mar 2006 11:33:54 -0600 13, 31 -- X-Server-Uuid: 8C3DB987-180B-4465-9446-45C15473FD3E 13, 31 -- Received: from sausexmb2.amd.com ([163.181.3.157]) by sausexbh1.amd.com 13, 31 -- with Microsoft SMTPSVC(6.0.3790.0); Thu, 23 Mar 2006 09:33:54 -0800 13, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 13, 31 -- Content-class: urn:content-classes:message 13, 31 -- MIME-Version: 1.0 13, 31 -- Subject: Career Opportunity (UTZ231) 13, 31 -- Date: Thu, 23 Mar 2006 11:33:52 -0600 13, 31 -- Message-ID: {4E9FB73E2965BD41888C731E554E248303FE7A05-at-SAUSEXMB2.amd.com} 13, 31 -- X-MS-Has-Attach: 13, 31 -- X-MS-TNEF-Correlator: 13, 31 -- Thread-Topic: Career Opportunity (UTZ231) 13, 31 -- Thread-Index: AcZOn/Qpsoao9sTUScSb8rBuPdvLBQ== 13, 31 -- From: "Gazda, Jerzy" {Jerzy.Gazda-at-ceriumlabs.com} 13, 31 -- To: Microscopy-at-microscopy.com 13, 31 -- X-OriginalArrivalTime: 23 Mar 2006 17:33:54.0463 (UTC) 13, 31 -- FILETIME=[F50C52F0:01C64E9F] 13, 31 -- X-WSS-ID: 683C03880SS101799-01-01 13, 31 -- Content-Type: text/plain; 13, 31 -- charset=us-ascii 13, 31 -- Content-Transfer-Encoding: 8bit 13, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2NHY5nv010885 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cprrrw-at-msn.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 23, 2006 at 09:49:39 ---------------------------------------------------------------------------
Email: cprrrw-at-msn.com Name: Patricia VanLuven
Organization: Home school
Education: K-8 Grade Grammar School
Location: Laingsburg, Michigan, USA
Question: Can you recommend a good protozoa identification book for an upper elementary/middle school student. Some are quite expensive and only available on-line (without preview), so I am looking for a recommendation before I make a selection. Thank you very much. Patty VanLuven
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Rainis, K.G. and Russell, B.J. 1996 Guide to Microlife 287pp, 5.5x8.5", paperback, $40.00. ISBN 0-531-11266-7 Franklin Watts, Danbury, CT. The price of this book is both unfortunate and understandable. Unfortunate, because it should be in the library of every class that studies the microlife of our environment; understandable, because almost every page has one or more excellent color light micrographs. It's a comprehensive field guide to the microworld. The authors make the statement that the 115 microorganisms described comprise 75-90% of those that may be encountered in the "wild". The habitats described are diverse: the home, soils, plants and debris, and four aquatic environments, with detailed advice on collecting methods for each. Described organisms are equally diverse, ranging from monerans to millimeter-sized arthropods. Species descriptions include ecological information, advice on collection and culture, and frequent suggestions for further investigation. Middle school - adult. RECOMMENDED
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 4, 18 -- From schooley-at-mcn.org Thu Mar 23 18:34:46 2006 4, 18 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2O0YkQU004675 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 18:34:46 -0600 4, 18 -- Received: from [66.42.18.121] (helo=[10.0.1.35]) 4, 18 -- by dns4.mcn.org with esmtpa (Exim 4.60) 4, 18 -- (envelope-from {schooley-at-mcn.org} ) 4, 18 -- id IWLW9X-000FIU-2S; Thu, 23 Mar 2006 16:34:46 -0800 4, 18 -- Mime-Version: 1.0 4, 18 -- Message-Id: {a06200701c048ed65586d-at-[10.0.1.35]} 4, 18 -- In-Reply-To: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- References: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- Date: Thu, 23 Mar 2006 16:32:23 -0800 4, 18 -- To: cprrrw-at-msn.com 4, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 4, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: protozoa identification book 4, 18 -- Cc: microscopy-at-microscopy.com 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
There is no "standard" way for the EM manufacturers to encode magnification or calibration in the image or image metadata. Some manufacturers store the information there with other tags, usually private, used to store the scale in various forms, other vendors omit this entirely and provide an additional text file with the information. Also, using "standard" TIF tags can be hazardous. For example, some text processing programs like Word use the x-calibration value (or y-calibration) for calculating the size of the image on paper. If you put in the real calibration there, you might end up wit Word trying to print the image at the real size, and you end up with a dot!
In order to get this information, you need to contact the manufacturer and ask them for the format that they are using. This may or may not be information that they can give you. If you have several instruments, you probably need to do this for each one.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- X-from: donc-at-asmicro.com [mailto:donc-at-asmicro.com] Sent: Thursday, March 23, 2006 10:06 AM To: Mike Bode
I am also interested to learn what methods are used in common brands of SEM to encode the scale or magnification information. We need this for our calibration and measurement software.
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 Headers============================== 3, 21 -- From donc-at-asmicro.com Thu Mar 23 11:04:34 2006 3, 21 -- Received: from smtp109.sbc.mail.re2.yahoo.com (smtp109.sbc.mail.re2.yahoo.com [68.142.229.96]) 3, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NH4XFi023944 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:04:33 -0600 3, 21 -- Received: (qmail 98485 invoked from network); 23 Mar 2006 17:04:31 -0000 3, 21 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.251.105.88 with login) 3, 21 -- by smtp109.sbc.mail.re2.yahoo.com with SMTP; 23 Mar 2006 17:04:30 -0000 3, 21 -- Message-ID: {015601c64e9b$63af7b60$c901a8c0-at-ASM11} 3, 21 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- To: "Microscopy List" {microscopy-at-microscopy.com} 3, 21 -- Subject: SEM digital image "metadata" 3, 21 -- Date: Thu, 23 Mar 2006 12:00:58 -0500 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: text/plain; 3, 21 -- charset="iso-8859-1" 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Priority: 3 3, 21 -- X-MSMail-Priority: Normal 3, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 3, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
==============================Original Headers============================== 13, 23 -- From Mike.Bode-at-soft-imaging.net Thu Mar 23 18:37:13 2006 13, 23 -- Received: from mail.soft-imaging.de (mail.soft-imaging.de [62.180.61.131]) 13, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2O0bCMh007749 13, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 18:37:12 -0600 13, 23 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 13, 23 -- by mail.soft-imaging.de (8.11.6/8.11.6/SuSE Linux 0.5) with ESMTP id k2O0bBD21654 13, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 01:37:11 +0100 13, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 23 -- Content-class: urn:content-classes:message 13, 23 -- MIME-Version: 1.0 13, 23 -- Content-Type: text/plain; 13, 23 -- charset="us-ascii" 13, 23 -- Subject: RE: [Microscopy] SEM digital image "metadata" 13, 23 -- Date: Fri, 24 Mar 2006 01:34:25 +0100 13, 23 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC94124FC3-at-ms-s-gws.soft-imaging.net} 13, 23 -- X-MS-Has-Attach: 13, 23 -- X-MS-TNEF-Correlator: 13, 23 -- Thread-Topic: [Microscopy] SEM digital image "metadata" 13, 23 -- Thread-Index: AcZOnBWMu/lqqZU6SrOOFRqh3pNidAAPbyog 13, 23 -- From: "Mike Bode" {Mike.Bode-at-soft-imaging.net} 13, 23 -- To: {Microscopy-at-microscopy.com} 13, 23 -- Content-Transfer-Encoding: 8bit 13, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2O0bCMh007749 ==============================End of - Headers==============================
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Email: mmanikka-at-umich.edu Name: Mohan Manikkam
Organization: University of Michigan
Title-Subject: [Filtered] Kodak NTB emulsion for autoradiography of in situ hybridized sections
Question: I am looking for the protocol and the experience of researchers with the Kodak NTB emulsion. I have used the previous versions, NTB-2 and NTB-3 but like to know if anyone has good results with the newer NTB emulsion with in situ hybridized tissue sections. I would appreciate members' valuable opinion and comment. Thanks.
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 Bplowman-at-pacific.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: Bplowman-at-pacific.edu Name: Barbara Plowman
Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry
Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
If you open these dysfunctional TIF files with Photoshop, it will gloriously delete all the abnormal header info.
Zeiss has their own format which is deleted when the image is converted from indexed color to grey scale.
gary g.
At 09:13 AM 3/23/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Thu Mar 23 20:10:41 2006 10, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2O2AfcF011966 10, 20 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 20:10:41 -0600 10, 20 -- Received: (qmail 14873 invoked from network); 23 Mar 2006 18:10:02 -0800 10, 20 -- Received: by simscan 1.1.0 ppid: 14870, pid: 14871, t: 0.1955s 10, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 20 -- by qsmtp4 with SMTP; 23 Mar 2006 18:10:02 -0800 10, 20 -- Message-Id: {6.2.3.4.2.20060323180759.023c98c0-at-mail.calweb.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 10, 20 -- Date: Thu, 23 Mar 2006 18:10:43 -0800 10, 20 -- To: michael-at-Shaffer.net 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] RE: SEM digital image "metadata" 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200603231713.k2NHDOgT004287-at-ns.microscopy.com} 10, 20 -- References: {200603231713.k2NHDOgT004287-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
briefly, I can tell you I am using acetonitrile (AN) as the "intermedium" after EtOH dehydration and embedding (as a substitute for PO) for now more than 15 years (human material, diagnostic and research specimens) without major problems in tissue &/or resin quality (polymerisation, cutting properties, stainability, stability in TEM-beam), provided you are aware of some specific properties of Acetonitrile.
As to my knowledge (and this was the cause for using AN instead of PO) Acetonitrile is stated "non-carcinogenic", despite being considered a mutagenic and cell toxic substance (PO is classified as "carcinogenic").
AN to 100% is water-miscible (so -theoretically- one should be able to use it as a substitute for EtOH as the dehydration solvent, but I've never tested that),
at ambient room and working conditions (humidity should not be too high, ventilated area needed like fume cupboard) you should get similar results for your specimen preparations, especially animal or human tissues (--} this was not the fact when I tried using so called "rapid dehydration methods" like the "acidified 2,2-DMP"-technique).
Vapor pressure of PO (unmiscible with water) is very high (-at-20 degr.C 588 hPa, -at- 33 degr.C 980 hPa, compared to AN: -at- 20 degr.C only 97 hPa; boiling point for PO: 35 degr.C, AN: 81 degr.C), "Highly flammable, vapors noxious, toxic if inhalated, swallowed or when contaminating skin"; MWC(1989): 40 ml/m3 - 70 mg/m3; Fresh water toxicity: Class 2 (do not waste into canalisation), but you will find all necessary physical data in the MSDS's provided with the substance delivered (hopefully!) (;:-))
for example see: http://www.jtbaker.com/msds/englishhtml/a0518.htm
(this was the } first { result at google)....you certainly will be "SHOCKED" but IMO: most of the chemicals used in (T)EM preparation do have some health risks if we don't work with them properly........(compare for that the statement of car producers: "Do not connect your exhaust pipe with the passenger room: don't inhale exhaust vapor...it might be lethal!") . Due to this "big" difference in vapor pressure, not only the substance's odours are "pleasant" as compared with PO.
Also, drying out of specimens during transfer of tissue into infiltration steps and pure resin (especially smallest ones) is not an issue any more. USE and Disposal of used solvent according to federal, national laws (in Europe/EC e.g. as "organic, non halogenated waste").
So - IMO - the most important thing: Using AN, you should be aware of a slower evaporation of solvent out of the tissue during infiltration (especially if room temperature is low) - thus you should
i) use specimen rotator(s)
ii) placing } infiltration { cups perhaps below a lamp (e.g. 60 W, at a distance of ca. 15 cm, temperature near specimens should be about 20-25 degr.C, see also v) below)
iii) placing specimens for "infiltration"-steps in flat "receptacula" (instead of [glass-] vials with a narrow neck and height of about 3 cm). For that purpose I fabricated on my own "special" infiltration silicone-rubber mo(u)lds (diam. ca. 1 cm, depth ca. 0.6 cm) which IMO are optimal for unhindered evaporation of the solvent, leaving also the option to polymerize the (pure) resin-fractions used for the infiltration-steps without any problem
iv) testing optimal infiltration times for the tissue you are embedding (usually, also for the big specimen blocks [up to 5 x 4 x 1 mm] I am working with, at least 45 min each infiltration step is -due to my experience - sufficient)
v) I use the following procedure (standardized for the diagnostic specimens, use of a specimen/probe rotator): - dehydration: ascending EtOH-series (50, 70, 80, 90, 96, 96, 100, 100, 100% EtOH),
- intermedium: 3 x pure AN (5, 10, 15 min), in ca. 5 ml snap cap vials, cap always closed (always take care of a surrounding humidity not to high !)
AN:Resin = 1: 1 : 1 x 45 minutes (at least), but that step can be elongated without problems up to 20 hours (i.e. e.g. over night, caps closed!), - before transfer into pure resin (into the infiltration moulds mentioned above, lamp) the cap of the vials is removed for at least 20-30 minutes (specimen rotator, under a lamp, see above),
pure RESIN: 3 x for at least 45 min each (use of lamp), take care of a humidity not to high !
There have been some papers related to the use of AN as a dehydration agent (and as a "safer" alternative to PO), if I remember correctly, in the 80ies or 90ies....if I find those or any in my files, I should be glad to share those informations with you (will take perhaps some hours of searching). One I have found by goo?gling: (http://www.emsdiasum.com/microscopy/technical/techtips/techtips2.aspx ) scroll down to # 18. Acetonitrile is a Safer, Better Dehydrating Solvent for Transmission Electron Microscopy: Propylene Oxide and Ethanol are commonly used dehydrating solvents for processing tissues for electron microscopy. But both solvents, however, have some undesirable properties: they are highly flammable, volatile, toxic and potentially carcinogenic. Acetonitrile is a direct substitution for ethanol and propylene oxide and it is safer to use and requires shorter dehydration times. It is freely miscible with water, alcohol, acetone and epoxy resin and it does not interfere with epoxy polymerization. Harold H. Edwards, Yu-Yan Yeh, Betty I. Tarnowsky, and Gregory R. Schonbaum. (1992). Acetonitrile as a Substitute for Ethanol/Propylene Oxide in Tissue Processing for Transmission Electron Microscopy: Comparison of Fine Structure and Lipid Solubility in Mouse Liver, Kidney, and Intestine. Microsc. Res. and Technique Vol. 21, pg.39-50
Hope this helps for now,
best regards and to all Listers: a beautiful Friday....weekend is coming (:-))
Wolfgang Muss
OR Dr. Wolfgang Muss EM-Lab =with pride: 25 years in operation by 2nd of Feb.2006= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at ------------------------------------------------------------------------ --------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------- Forthcoming Meetings:
SECOND ANNOUNCEMENT, REGISTER NOW ONLINE at: http://www.scur.org.pl
33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl -------------------------------- 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic -------------------------------- 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both Bplowman-at-pacific.edu as well as the MIcroscopy Listserver --------------------------------------------------------------------------- Email: Bplowman-at-pacific.edu Name: Barbara Plowman Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
} There is no "standard" way for the EM manufacturers to encode } magnification or calibration in the image or image metadata. } Some manufacturers store the information there with other } tags, usually private, used to store the scale in various } forms, other vendors omit this entirely and provide an } additional text file with the information.
Which begs the question "Why is there no standard method?" There is certainly enough EXIF fields still available. It would seem all that is needed is the push and for us to come up with a minimal number of field designations to apphoach the EXIF (or IPTC) people with.
} Also, using "standard" TIF tags can be hazardous. For } example, some text processing programs like Word use the } x-calibration value (or } y-calibration) for calculating the size of the image on } paper. If you put in the real calibration there, you might } end up wit Word trying to print the image at the real size, } and you end up with a dot!
Yes ... It's difficult enough to know why the EM manufacturers do not put the correct resolution into the file such that it'll print at the correct size. However, this TIFF field is not what I speaking of.
} In order to get this information, you need to contact the } manufacturer and ask them for the format that they are using. } This may or may not be information that they can give you. If } you have several instruments, you probably need to do this } for each one.
Personally, I have no need to do it for any other than my own SEM. However, if I am successful with the code, I'll get back to the group with the example. I know that at least a couple of SEM manufacturers are similar, if not the same.
Hello Everyone, I just purchased a copy of "Guide to Microlife" (how could I resist not looking at pond water and knowing what I'm looking at!) from Buy.com for under $24 including shipping. As an aside, they offered my a $25.00 discount coupon for my next order. One might consider ordering one book, then with the coupon order more...
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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schooley-at-mcn.org To: frank.karl-at-degussa.com 03/23/2006 07:36 cc: PM Subject: [Microscopy] Re: AskAMicroscopist: protozoa identification book Please respond to schooley
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} } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (cprrrw-at-msn.com) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Thursday, March 23, 2006 at 09:49:39 } ---------------------------------------------------------------------------
} } Email: cprrrw-at-msn.com } Name: Patricia VanLuven } } Organization: Home school } } Education: K-8 Grade Grammar School } } Location: Laingsburg, Michigan, USA } } Question: Can you recommend a good protozoa identification book for } an upper elementary/middle school student. Some are quite expensive } and only available on-line (without preview), so I am looking for a } recommendation before I make a selection. Thank you very much. } Patty VanLuven } } ---------------------------------------------------------------------------
Patricia - You may consider this too expensive, but it's what you need; maybe you can find a used copy. The description is taken from the MICRO bibliography (URL below).
Rainis, K.G. and Russell, B.J. 1996 Guide to Microlife 287pp, 5.5x8.5", paperback, $40.00. ISBN 0-531-11266-7 Franklin Watts, Danbury, CT. The price of this book is both unfortunate and understandable. Unfortunate, because it should be in the library of every class that studies the microlife of our environment; understandable, because almost every page has one or more excellent color light micrographs. It's a comprehensive field guide to the microworld. The authors make the statement that the 115 microorganisms described comprise 75-90% of those that may be encountered in the "wild". The habitats described are diverse: the home, soils, plants and debris, and four aquatic environments, with detailed advice on collecting methods for each. Described organisms are equally diverse, ranging from monerans to millimeter-sized arthropods. Species descriptions include ecological information, advice on collection and culture, and frequent suggestions for further investigation. Middle school - adult. RECOMMENDED
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 4, 18 -- From schooley-at-mcn.org Thu Mar 23 18:34:46 2006 4, 18 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2O0YkQU004675 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 18:34:46 -0600 4, 18 -- Received: from [66.42.18.121] (helo=[10.0.1.35]) 4, 18 -- by dns4.mcn.org with esmtpa (Exim 4.60) 4, 18 -- (envelope-from {schooley-at-mcn.org} ) 4, 18 -- id IWLW9X-000FIU-2S; Thu, 23 Mar 2006 16:34:46 -0800 4, 18 -- Mime-Version: 1.0 4, 18 -- Message-Id: {a06200701c048ed65586d-at-[10.0.1.35]} 4, 18 -- In-Reply-To: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- References: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- Date: Thu, 23 Mar 2006 16:32:23 -0800 4, 18 -- To: cprrrw-at-msn.com 4, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 4, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: protozoa identification book 4, 18 -- Cc: microscopy-at-microscopy.com 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 29, 18 -- From frank.karl-at-degussa.com Fri Mar 24 07:48:26 2006 29, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 29, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2ODmOCH020633 29, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Mar 2006 07:48:25 -0600 29, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 29, 18 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2ODmHpE021422; 29, 18 -- Fri, 24 Mar 2006 14:48:18 +0100 29, 18 -- In-Reply-To: {200603240036.k2O0ac4B007038-at-ns.microscopy.com} 29, 18 -- Subject: Re: AskAMicroscopist: protozoa identification book 29, 18 -- To: schooley-at-mcn.org, microscopy-at-msa.microscopy.com 29, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 29, 18 -- Message-ID: {OF46EF54FD.85491A20-ON8525713B.004B5C62-8525713B.004BD1C7-at-degussa.com} 29, 18 -- From: frank.karl-at-degussa.com 29, 18 -- Date: Fri, 24 Mar 2006 08:48:09 -0500 29, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 29, 18 -- 03/24/2006 07:48:20 AM 29, 18 -- MIME-Version: 1.0 29, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
The "Guide to Microlife" is $23 from amazon.com. A better, but more expensive (but without all the color photographs) is Theodore Jahn, et al. "How to Know the Protozoa". The classification is outdated, since its 1978, but it's still a useful guide. Should be available used from Advanced Book Exchange (abebooks.com), Powell's, and the like. But be careful! Many of the used copies are the 1948 or 1963 editions. Better still is Patterson's "Free-living Freshwater Protozoa: A Color Guide", but this is $60. Unfortunately, that's cheap anymore for books.
Phil
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-- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Fri Mar 24 08:15:56 2006 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2OEFt49030656 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 08:15:55 -0600 4, 22 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k2OEpR4v008018 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 09:53:36 -0500 4, 22 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 22 -- Fri, 24 Mar 2006 09:08:27 -0500 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230901c049a9a79f16-at-[141.209.160.132]} 4, 22 -- In-Reply-To: {200603240040.k2O0exLx020415-at-ns.microscopy.com} 4, 22 -- References: {200603240040.k2O0exLx020415-at-ns.microscopy.com} 4, 22 -- Date: Fri, 24 Mar 2006 09:08:39 -0500 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: [Microscopy] Re: AskAMicroscopist: protozoa identification book 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 24 Mar 2006 14:08:27.0851 (UTC) FILETIME=[6C3A59B0:01C64F4C] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -4 () L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
The Minnesota Microscopy Society (MMS) will hold its annual Spring Symposium on April 21st. This year's topic is "Microcopy in Failure Analysis". The all day event will be held at the Minnesota Science Museum.
Topic will include:
Plastic Component Failure Analysis Failure Analysis in the 21st Century, Nano-Scale Materials Specimen Selection in Microscopy for Failure Analysis Use of the SEM in the Failure Analysis of Cardiac Pacing Leads Vendor Displays
Cost $75 per person for memember and $85 per person for non members. Lunch and coffee breaks provided.
Reservations MUST be made no later than Friday, April 14th. Register by e-mailing Bede Willenbring at Bede.Willenbring-at-hbfuller.com, or by phone at 651-236-5470. Include your name, company, phone number, and e-mail address.
Full details are available online at http://www.MNmicroscopy.org - just click on the "current newsletter" in your preferred format; html for browsing or pdf for printing out to show your colleagues.
Thank you
Minnesota Microscopy Society
Prof. Stuart McKernan IT Characterization Facility, University of Minnesota E-mail : stuartm-at-umn.edu 12 Shepherd Labs, Office: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 626-7594
==============================Original Headers============================== 13, 15 -- From stuartm-at-umn.edu Fri Mar 24 09:22:57 2006 13, 15 -- Received: from mtaout-w.tc.umn.edu (mtaout-w.tc.umn.edu [160.94.160.21]) 13, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2OFMvoG009757 13, 15 -- for {microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 09:22:57 -0600 13, 15 -- Received: from [160.94.16.136] (cfsigma.charfac.umn.edu [160.94.16.136]) by mtaout-w.tc.umn.edu with ESMTP for microscopy-at-microscopy.com; Fri, 24 Mar 2006 09:22:49 -0600 (CST) 13, 15 -- X-Umn-Remote-Mta: [N] cfsigma.charfac.umn.edu [160.94.16.136] #+LO+TS+AU+HN 13, 15 -- Mime-Version: 1.0 (Apple Message framework v746.3) 13, 15 -- Content-Transfer-Encoding: 7bit 13, 15 -- Message-Id: {E6603809-4267-4B31-8C2F-132ED229FE51-at-umn.edu} 13, 15 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 13, 15 -- To: microscopy-at-microscopy.com 13, 15 -- From: stuart McKernan {stuartm-at-umn.edu} 13, 15 -- Subject: MMS Spring Symposium 13, 15 -- Date: Fri, 24 Mar 2006 09:22:47 -0600 13, 15 -- X-Mailer: Apple Mail (2.746.3) ==============================End of - Headers==============================
Sir, it is my duty, to distribute this your valued information to the MSA-listers. I am sorry if I "encouraged" somebody to use that stuff without cautions you are stating below. I always adhered to GLP procedures, in this special case by using } fume cupboard {. Also I found a hint on the toxic effects of ACN(Acetonitrile) when combined with water in the List Server Archives, July 1994 by Marcelle A Gillott, who stated: *** CAUTION *** acetonitrile combined with water releases hydrogen cyanide gas !!!
while it is touted as being considerably less toxic than PO users should be aware of the above reaction if it is being used as a dehydrant.
I shall use ACN from now on more carefully than ever.
Thank you for your highly valued comment !
Best regards Wolfgang Muss
PS: another issue for people working with fixatives - similar to this "hidden ACN problem" - would be formaldehyde (as used in tissue fixation procedures) and traces of hydrochloric acid ==} producing highly toxic and cancerogenic Bis-Chloromethylether (bis-CME)....
First of all, I do not disagree with any of your statements on the safety comparison between acetonitrile and propylene oxide.
However I do feel that you have subsequently understated the hazards of acetonitrile.
Acetonitrile (ACN) and Acrylonitrile (AN) are both significantly hazardous chemicals in that they break down in the body and generate hydrogen cyanide, HCN. Both AN and ACN are readily absorbed through skin and tissues. Cyanide poisoning has many stages, none of which are good! Please be careful with these very useful, but hazardous solvents.
As you have previously mentioned, Use of these solvents in your lab should be done carefully, and firstly with SOPs and engineering controls well established and implemented. Also with resulting PPE verified, and employed. Depending on the quantities and concentrations, you may want to consider keeping one of several HCN antidotes on hand, or make your emergency response system aware of your use of these chemicals so that they can have the antidote on hand. Many fire departments and paramedics units carry HCN antidotes or have a special first-aid treatment for victims exposed to chemicals in this family.
Please communicate this info to your colleagues working with these chemicals, and by all means consult the MSDS for every chemical that you plan to use.
Brad Huggins BP Chemicals Naperville, IL
Sometimes, being careful
Some more info below:
Response of Humans to HCN in Air 270 ppm Immediately fatal 181 ppm Fatal after 10 minutes 135 ppm Fatal after 30 minutes 110-135 ppm Fatal 30-60+ minutes 45-55 ppm Tolerated for 30-60 min
Early Physical Findings with Contaminated Victim: Special caution for Head, Ear, Eye, Nose and Mouth/Throat Bright red retinal veins and arteries Smell of bitter almonds on the breath
Cyanide antidotes if diagnosis is certain: Sodium nitrite intravenously Sodium thiosulfite intravenously Kelocyanor available in UK and France Amyl nitrite ampoules: temporizing therapy until IV access is obtained
Subchronic Toxicity Sporadic vapor exposure for 6 years: Loss of appetite, nervousness, vertigo, headache, nausea, vomiting Goldsmith apprentice: Headache, listlessness, numbness, partial paralysis of left arm and leg, partial loss of vision left eye, and EKG abnormalities
-----Original Message----- X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at] Sent: Friday, March 24, 2006 3:19 AM To: Huggins, Bradley J
Good morning, dear Barbara,
briefly, I can tell you I am using acetonitrile (AN) as the "intermedium" after EtOH dehydration and embedding (as a substitute for PO) for now more than 15 years (human material, diagnostic and research specimens) without major problems in tissue &/or resin quality (polymerisation, cutting properties, stainability, stability in TEM-beam), provided you are aware of some specific properties of Acetonitrile.
As to my knowledge (and this was the cause for using AN instead of PO) Acetonitrile is stated "non-carcinogenic", despite being considered a mutagenic and cell toxic substance (PO is classified as "carcinogenic").
AN to 100% is water-miscible (so -theoretically- one should be able to use it as a substitute for EtOH as the dehydration solvent, but I've never tested that),
at ambient room and working conditions (humidity should not be too high, ventilated area needed like fume cupboard) you should get similar results for your specimen preparations, especially animal or human tissues (--} this was not the fact when I tried using so called "rapid dehydration methods" like the "acidified 2,2-DMP"-technique).
Vapor pressure of PO (unmiscible with water) is very high (-at-20 degr.C 588 hPa, -at- 33 degr.C 980 hPa, compared to AN: -at- 20 degr.C only 97 hPa; boiling point for PO: 35 degr.C, AN: 81 degr.C), "Highly flammable, vapors noxious, toxic if inhalated, swallowed or when contaminating skin"; MWC(1989): 40 ml/m3 - 70 mg/m3; Fresh water toxicity: Class 2 (do not waste into canalisation), but you will find all necessary physical data in the MSDS's provided with the substance delivered (hopefully!) (;:-))
for example see: http://www.jtbaker.com/msds/englishhtml/a0518.htm
(this was the } first { result at google)....you certainly will be "SHOCKED" but IMO: most of the chemicals used in (T)EM preparation do have some health risks if we don't work with them properly........(compare for that the statement of car producers: "Do not connect your exhaust pipe with the passenger room: don't inhale exhaust vapor...it might be lethal!") . Due to this "big" difference in vapor pressure, not only the substance's odours are "pleasant" as compared with PO.
Also, drying out of specimens during transfer of tissue into infiltration steps and pure resin (especially smallest ones) is not an issue any more. USE and Disposal of used solvent according to federal, national laws (in Europe/EC e.g. as "organic, non halogenated waste").
So - IMO - the most important thing: Using AN, you should be aware of a slower evaporation of solvent out of the tissue during infiltration (especially if room temperature is low) - thus you should
i) use specimen rotator(s)
ii) placing } infiltration { cups perhaps below a lamp (e.g. 60 W, at a distance of ca. 15 cm, temperature near specimens should be about 20-25 degr.C, see also v) below)
iii) placing specimens for "infiltration"-steps in flat "receptacula" (instead of [glass-] vials with a narrow neck and height of about 3 cm).
For that purpose I fabricated on my own "special" infiltration silicone-rubber mo(u)lds (diam. ca. 1 cm, depth ca. 0.6 cm) which IMO are optimal for unhindered evaporation of the solvent, leaving also the option to polymerize the (pure) resin-fractions used for the infiltration-steps without any problem
iv) testing optimal infiltration times for the tissue you are embedding (usually, also for the big specimen blocks [up to 5 x 4 x 1 mm] I am working with, at least 45 min each infiltration step is -due to my experience - sufficient)
v) I use the following procedure (standardized for the diagnostic specimens, use of a specimen/probe rotator): - dehydration: ascending EtOH-series (50, 70, 80, 90, 96, 96, 100, 100, 100% EtOH),
- intermedium: 3 x pure AN (5, 10, 15 min), in ca. 5 ml snap cap vials, cap always closed (always take care of a surrounding humidity not to high !)
AN:Resin = 1: 1 : 1 x 45 minutes (at least), but that step can be elongated without problems up to 20 hours (i.e. e.g. over night, caps closed!), - before transfer into pure resin (into the infiltration moulds mentioned above, lamp) the cap of the vials is removed for at least 20-30 minutes (specimen rotator, under a lamp, see above),
pure RESIN: 3 x for at least 45 min each (use of lamp), take care of a humidity not to high !
There have been some papers related to the use of AN as a dehydration agent (and as a "safer" alternative to PO), if I remember correctly, in the 80ies or 90ies....if I find those or any in my files, I should be glad to share those informations with you (will take perhaps some hours of searching). One I have found by goo?gling: (http://www.emsdiasum.com/microscopy/technical/techtips/techtips2.aspx ) scroll down to # 18. Acetonitrile is a Safer, Better Dehydrating Solvent for Transmission Electron Microscopy: Propylene Oxide and Ethanol are commonly used dehydrating solvents for processing tissues for electron microscopy. But both solvents, however, have some undesirable properties: they are highly flammable, volatile, toxic and potentially carcinogenic. Acetonitrile is a direct substitution for ethanol and propylene oxide and it is safer to use and requires shorter dehydration times. It is freely miscible with water, alcohol, acetone and epoxy resin and it does not interfere with epoxy polymerization. Harold H. Edwards, Yu-Yan Yeh, Betty I. Tarnowsky, and Gregory R. Schonbaum. (1992). Acetonitrile as a Substitute for Ethanol/Propylene Oxide in Tissue Processing for Transmission Electron Microscopy: Comparison of Fine Structure and Lipid Solubility in Mouse Liver, Kidney, and Intestine. Microsc. Res. and Technique Vol. 21, pg.39-50
Hope this helps for now,
best regards and to all Listers: a beautiful Friday....weekend is coming (:-))
Wolfgang Muss
OR Dr. Wolfgang Muss EM-Lab =with pride: 25 years in operation by 2nd of Feb.2006= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at ------------------------------------------------------------------------ --------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------ - Forthcoming Meetings:
SECOND ANNOUNCEMENT, REGISTER NOW ONLINE at: http://www.scur.org.pl
33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl -------------------------------- 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic -------------------------------- 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
---- 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 Bplowman-at-pacific.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ --- Email: Bplowman-at-pacific.edu Name: Barbara Plowman Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or
acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
Just about all procedures using chemicals commonly found in electron microscopy should be performed in fume hoods. All scientists and support staff should read the appropriate Material Safety Data Sheets (MSDS) for each chemical used (many online sites). Save copies of these for future reference/emergencies/etc. and update every several years. A 10-15 year old MSDS may be useless.
Of course there will be exceptions but use "common sense" [:)]. Expect your safety guy/gal to lack this quality.
Even old dogs learn new tricks!
X-from the New Orleans Diaspora,
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, MSA/SRRC SWSRU P.O. Box 350 Stoneville, MS 38776
} } } {W.Muss-at-salk.at} 03/24/06 11:41AM } } } ---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Sir, it is my duty, to distribute this your valued information to the MSA-listers. I am sorry if I "encouraged" somebody to use that stuff without cautions you are stating below. I always adhered to GLP procedures, in this special case by using } fume cupboard {. Also I found a hint on the toxic effects of ACN(Acetonitrile) when combined with water in the List Server Archives, July 1994 by Marcelle A Gillott, who stated: *** CAUTION *** acetonitrile combined with water releases hydrogen cyanide gas !!!
while it is touted as being considerably less toxic than PO users should be aware of the above reaction if it is being used as a dehydrant.
I shall use ACN from now on more carefully than ever.
Thank you for your highly valued comment !
Best regards Wolfgang Muss
PS: another issue for people working with fixatives - similar to this "hidden ACN problem" - would be formaldehyde (as used in tissue fixation procedures) and traces of hydrochloric acid ==} producing highly toxic and cancerogenic Bis-Chloromethylether (bis-CME)....
First of all, I do not disagree with any of your statements on the safety comparison between acetonitrile and propylene oxide.
However I do feel that you have subsequently understated the hazards of acetonitrile.
Acetonitrile (ACN) and Acrylonitrile (AN) are both significantly hazardous chemicals in that they break down in the body and generate hydrogen cyanide, HCN. Both AN and ACN are readily absorbed through skin and tissues. Cyanide poisoning has many stages, none of which are good! Please be careful with these very useful, but hazardous solvents.
As you have previously mentioned, Use of these solvents in your lab should be done carefully, and firstly with SOPs and engineering controls well established and implemented. Also with resulting PPE verified, and employed. Depending on the quantities and concentrations, you may want to consider keeping one of several HCN antidotes on hand, or make your emergency response system aware of your use of these chemicals so that they can have the antidote on hand. Many fire departments and paramedics units carry HCN antidotes or have a special first-aid treatment for victims exposed to chemicals in this family.
Please communicate this info to your colleagues working with these chemicals, and by all means consult the MSDS for every chemical that you plan to use.
Brad Huggins BP Chemicals Naperville, IL
Sometimes, being careful
Some more info below:
Response of Humans to HCN in Air 270 ppm Immediately fatal 181 ppm Fatal after 10 minutes 135 ppm Fatal after 30 minutes 110-135 ppm Fatal 30-60+ minutes 45-55 ppm Tolerated for 30-60 min
Early Physical Findings with Contaminated Victim: Special caution for Head, Ear, Eye, Nose and Mouth/Throat Bright red retinal veins and arteries Smell of bitter almonds on the breath
Cyanide antidotes if diagnosis is certain: Sodium nitrite intravenously Sodium thiosulfite intravenously Kelocyanor available in UK and France Amyl nitrite ampoules: temporizing therapy until IV access is obtained
Subchronic Toxicity Sporadic vapor exposure for 6 years: Loss of appetite, nervousness, vertigo, headache, nausea, vomiting Goldsmith apprentice: Headache, listlessness, numbness, partial paralysis of left arm and leg, partial loss of vision left eye, and EKG abnormalities
-----Original Message----- X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at] Sent: Friday, March 24, 2006 3:19 AM To: Huggins, Bradley J
Good morning, dear Barbara,
briefly, I can tell you I am using acetonitrile (AN) as the "intermedium" after EtOH dehydration and embedding (as a substitute for PO) for now more than 15 years (human material, diagnostic and research specimens) without major problems in tissue &/or resin quality (polymerisation, cutting properties, stainability, stability in TEM-beam), provided you are aware of some specific properties of Acetonitrile.
As to my knowledge (and this was the cause for using AN instead of PO) Acetonitrile is stated "non-carcinogenic", despite being considered a mutagenic and cell toxic substance (PO is classified as "carcinogenic").
AN to 100% is water-miscible (so -theoretically- one should be able to use it as a substitute for EtOH as the dehydration solvent, but I've never tested that),
at ambient room and working conditions (humidity should not be too high, ventilated area needed like fume cupboard) you should get similar results for your specimen preparations, especially animal or human tissues (--} this was not the fact when I tried using so called "rapid dehydration methods" like the "acidified 2,2-DMP"-technique).
Vapor pressure of PO (unmiscible with water) is very high (-at-20 degr.C 588 hPa, -at- 33 degr.C 980 hPa, compared to AN: -at- 20 degr.C only 97 hPa; boiling point for PO: 35 degr.C, AN: 81 degr.C), "Highly flammable, vapors noxious, toxic if inhalated, swallowed or when contaminating skin"; MWC(1989): 40 ml/m3 - 70 mg/m3; Fresh water toxicity: Class 2 (do not waste into canalisation), but you will find all necessary physical data in the MSDS's provided with the substance delivered (hopefully!) (;:-))
for example see: http://www.jtbaker.com/msds/englishhtml/a0518.htm
(this was the } first { result at google)....you certainly will be "SHOCKED" but IMO: most of the chemicals used in (T)EM preparation do have some health risks if we don't work with them properly........(compare for that the statement of car producers: "Do not connect your exhaust pipe with the passenger room: don't inhale exhaust vapor...it might be lethal!") . Due to this "big" difference in vapor pressure, not only the substance's odours are "pleasant" as compared with PO.
Also, drying out of specimens during transfer of tissue into infiltration steps and pure resin (especially smallest ones) is not an issue any more. USE and Disposal of used solvent according to federal, national laws (in Europe/EC e.g. as "organic, non halogenated waste").
So - IMO - the most important thing: Using AN, you should be aware of a slower evaporation of solvent out of the tissue during infiltration (especially if room temperature is low) - thus you should
i) use specimen rotator(s)
ii) placing } infiltration { cups perhaps below a lamp (e.g. 60 W, at a distance of ca. 15 cm, temperature near specimens should be about 20-25 degr.C, see also v) below)
iii) placing specimens for "infiltration"-steps in flat "receptacula" (instead of [glass-] vials with a narrow neck and height of about 3 cm).
For that purpose I fabricated on my own "special" infiltration silicone-rubber mo(u)lds (diam. ca. 1 cm, depth ca. 0.6 cm) which IMO are optimal for unhindered evaporation of the solvent, leaving also the option to polymerize the (pure) resin-fractions used for the infiltration-steps without any problem
iv) testing optimal infiltration times for the tissue you are embedding (usually, also for the big specimen blocks [up to 5 x 4 x 1 mm] I am working with, at least 45 min each infiltration step is -due to my experience - sufficient)
v) I use the following procedure (standardized for the diagnostic specimens, use of a specimen/probe rotator): - dehydration: ascending EtOH-series (50, 70, 80, 90, 96, 96, 100, 100, 100% EtOH),
- intermedium: 3 x pure AN (5, 10, 15 min), in ca. 5 ml snap cap vials, cap always closed (always take care of a surrounding humidity not to high !)
AN:Resin = 1: 1 : 1 x 45 minutes (at least), but that step can be elongated without problems up to 20 hours (i.e. e.g. over night, caps closed!), - before transfer into pure resin (into the infiltration moulds mentioned above, lamp) the cap of the vials is removed for at least 20-30 minutes (specimen rotator, under a lamp, see above),
pure RESIN: 3 x for at least 45 min each (use of lamp), take care of a humidity not to high !
There have been some papers related to the use of AN as a dehydration agent (and as a "safer" alternative to PO), if I remember correctly, in the 80ies or 90ies....if I find those or any in my files, I should be glad to share those informations with you (will take perhaps some hours of searching). One I have found by goo?gling: (http://www.emsdiasum.com/microscopy/technical/techtips/techtips2.aspx ) scroll down to # 18. Acetonitrile is a Safer, Better Dehydrating Solvent for Transmission Electron Microscopy: Propylene Oxide and Ethanol are commonly used dehydrating solvents for processing tissues for electron microscopy. But both solvents, however, have some undesirable properties: they are highly flammable, volatile, toxic and potentially carcinogenic. Acetonitrile is a direct substitution for ethanol and propylene oxide and it is safer to use and requires shorter dehydration times. It is freely miscible with water, alcohol, acetone and epoxy resin and it does not interfere with epoxy polymerization. Harold H. Edwards, Yu-Yan Yeh, Betty I. Tarnowsky, and Gregory R. Schonbaum. (1992). Acetonitrile as a Substitute for Ethanol/Propylene Oxide in Tissue Processing for Transmission Electron Microscopy: Comparison of Fine Structure and Lipid Solubility in Mouse Liver, Kidney, and Intestine. Microsc. Res. and Technique Vol. 21, pg.39-50
Hope this helps for now,
best regards and to all Listers: a beautiful Friday....weekend is coming (:-))
Wolfgang Muss
OR Dr. Wolfgang Muss EM-Lab =with pride: 25 years in operation by 2nd of Feb.2006= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at ------------------------------------------------------------------------ --------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------ - Forthcoming Meetings:
SECOND ANNOUNCEMENT, REGISTER NOW ONLINE at: http://www.scur.org.pl
33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl -------------------------------- 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic -------------------------------- 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
---- 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 Bplowman-at-pacific.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ --- Email: Bplowman-at-pacific.edu Name: Barbara Plowman Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or
acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
I got a question I hope the collective wisdom can help answer. I have TiO2 in an organic matrix. PLM shows the expected optical properties, but I would like to run a microchemical test to confirm the presence of Ti. I seem to remember a bead test to fuse the TiO2 into something water soluble followed by a microchemical with ? quinoline and ammonium thiocyanate? Squaric acid?
Any suggestion to get my TiO2 into solution would be welcome!
Thanks!! Frank
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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==============================Original Headers============================== 9, 17 -- From frank.karl-at-degussa.com Fri Mar 24 13:36:48 2006 9, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 9, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2OJalsF010151 9, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Mar 2006 13:36:47 -0600 9, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 9, 17 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2OJaiue027149 9, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Mar 2006 20:36:45 +0100 9, 17 -- Subject: Question Microchemical test for Ti 9, 17 -- To: microscopy-at-msa.microscopy.com 9, 17 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 9, 17 -- Message-ID: {OF6F9B1405.4F061B37-ON8525713B.006A27A0-8525713B.006BB8D7-at-degussa.com} 9, 17 -- From: frank.karl-at-degussa.com 9, 17 -- Date: Fri, 24 Mar 2006 14:36:36 -0500 9, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 9, 17 -- 03/24/2006 01:36:46 PM 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (streiker-at-sbcglobal.net) from on Saturday, March 25, 2006 at 20:13:39 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both streiker-at-sbcglobal.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: streiker-at-sbcglobal.net Name: Scott Streiker
Organization: University of Dayton
Education: Graduate College
Location: Dayton, Ohio, USA
Title: Atomic lattice with TEM
Question: What is the protocol for using a TEM to view atomic lattice/planes of carbon nano tubes at direct magnification over 200K at accel voltage of 100kV or greater?
which seems to describe the use of potassium thiocarbonate.
I don't know if anyone uses ring oven techniques any more, they were widely developed and promoted by Phil West and his wife in Baton Rouge in the 1960s and 1970s and at the time seemed to me very interesting. Then I changed jobs.
cheers
rtch
On 24 Mar 2006 at 13:38, frank.karl-at-degussa.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 } ---------------------------------------------------------------------------- } } I got a question I hope the collective wisdom can help answer. I have TiO2 } in an organic matrix. PLM shows the expected optical properties, but I } would like to run a microchemical test to confirm the presence of Ti. I } seem to remember a bead test to fuse the TiO2 into something water soluble } followed by a microchemical with ? quinoline and ammonium thiocyanate? } Squaric acid? } } Any suggestion to get my TiO2 into solution would be welcome! } } Thanks!! Frank } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 }
-- 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============================== 13, 27 -- From r.sims-at-auckland.ac.nz Sun Mar 26 17:08:49 2006 13, 27 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) 13, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2QN8mro017444 13, 27 -- for {microscopy-at-msa.microscopy.com} ; Sun, 26 Mar 2006 17:08:49 -0600 13, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 13, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 57B9634D50; 13, 27 -- Mon, 27 Mar 2006 11:08:47 +1200 (NZST) 13, 27 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) 13, 27 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 13, 27 -- with ESMTP id 21609-16; Mon, 27 Mar 2006 11:08:47 +1200 (NZST) 13, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 13, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 0810934E65; 13, 27 -- Mon, 27 Mar 2006 11:08:45 +1200 (NZST) 13, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 13, 27 -- To: frank.karl-at-degussa.com 13, 27 -- Date: Mon, 27 Mar 2006 11:12:15 +1200 13, 27 -- MIME-Version: 1.0 13, 27 -- Subject: Re: [Microscopy] Question Microchemical test for Ti 13, 27 -- Cc: microscopy-at-msa.microscopy.com 13, 27 -- Message-ID: {4427C88F.2117.818086-at-localhost} 13, 27 -- Priority: normal 13, 27 -- In-reply-to: {200603241938.k2OJc0s8012169-at-ns.microscopy.com} 13, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 13, 27 -- Content-type: text/plain; charset=US-ASCII 13, 27 -- Content-transfer-encoding: 7BIT 13, 27 -- Content-description: Mail message body 13, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
Here are the happy-ending results of your wise advices for TC embedding, both in monolayer and pellet. Both methods worked well, with the advantage of monolayer embedding being that it is 1 day shorter (the penetration times are reduced) and keeps intact the cell contacts. The only disadvantage I found was by cutting, which requires either a lot of experience or a lot of patience (or both?). The cultures must be pretty confluent too, which is an evident limitation. I list the methods I found successful because they may be helpful for somebody else. I don’t want to favor a method for another, I just list what worked for me. This does not mean that other proposed methods are not valid!!
1. For pellet embedding, the good trick was to add the fixative in the culture medium at 37°C, wait 30 sec and scrape the cells, then pellet briefly in eppis at full speed, then replace with fresh fixative and continuing fixation at 4°C. 2. For monolayer embedding, the following method gave me entire satisfaction: do all steps in 3.5 cm petri dish, simply avoiding propyleneoxyde (mix epon with ethanol). Cover the cells with a thin layer of Epon (1 ml/3.5 cm petri dish, which gives approx. the same epon thickness as the bottom of the petri dish) and put BEEM capsules, whose tip has previously been cut out, upside-down in the resin. The next day, the BEEM capsules being embedded in the thin layer of Epon, fill them with Epon and cure for another 24 h. Next day, you can simply pull the capsules out of the petri dishes. It may happen that some plastic comes with the capsule, but never the entire surface, just choose a place without plastic to cut the pyramid.
I want to thank warmly all listers who helped me, and the others too because it wouldn’t be fair otherwise ;-)
Stephane-without-i
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==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Mon Mar 27 03:04:35 2006 7, 18 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2R94ZN9003966 7, 18 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 03:04:35 -0600 7, 18 -- Received: (qmail 42811 invoked by uid 60001); 27 Mar 2006 09:04:35 -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=sH4swVF+dgCyC1MJ5E++jtFHS8ZY/AaK2W1ufZY5MjlwnTEF0brCbLkJkz2TCNprhe4aoFUSRqK2H8Aste8wdFbPacSBD42sc2y/u+SL/NmUimpGePNyHdcumYJ5xW+x/9KopnUzGA8aUyRhoazBEZnhWPipN9rpGuj+/e0sATY= ; 7, 18 -- Message-ID: {20060327090435.42807.qmail-at-web37413.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Mon, 27 Mar 2006 01:04:35 PST 7, 18 -- Date: Mon, 27 Mar 2006 01:04:35 -0800 (PST) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: TC cell preparation: happy end and conclusion 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==============================
I am looking of an adhesive of very low viscosity that will take onto titanium. The purpose is for the preparation of cross-sectional TEM specimens. The M-bond 610 works well for almost everything else, except Ti. Can someone help me with advice please?.
Thanks
Basil
Dr. Basil Julies Head Electron Microscope Unit Physics Department University of the Western Cape Private Bag X17 Bellville 7535 Tel : (27)(21) 959 2327 or 959 3458 Fax : (27)(21) 959 1335 or 959 3474
==============================Original Headers============================== 5, 19 -- From bjulies-at-uwc.ac.za Mon Mar 27 09:20:26 2006 5, 19 -- Received: from uwcunx.uwc.ac.za (uwcunx.uwc.ac.za [196.11.235.8]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2RFKO9F001403 5, 19 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 09:20:25 -0600 5, 19 -- Received: (qmail 7298 invoked from network); 27 Mar 2006 14:27:13 -0000 5, 19 -- Received: from itsnw.uwc.ac.za (HELO Services-02.uwc.ac.za) (192.102.9.71) 5, 19 -- by uwcunx.uwc.ac.za with SMTP; 27 Mar 2006 14:27:13 -0000 5, 19 -- Received: from UWC-MTA by Services-02.uwc.ac.za 5, 19 -- with Novell_GroupWise; Mon, 27 Mar 2006 17:24:09 +0200 5, 19 -- Message-Id: {s4281fb9.005-at-Services-02.uwc.ac.za} 5, 19 -- X-Mailer: Novell GroupWise Internet Agent 6.5.2 5, 19 -- Date: Mon, 27 Mar 2006 17:23:37 +0200 5, 19 -- From: "Basil Julies" {bjulies-at-uwc.ac.za} 5, 19 -- To: {microscopy-at-microscopy.com} 5, 19 -- Subject: adhesive for Titanium 5, 19 -- Mime-Version: 1.0 5, 19 -- Content-Type: text/plain; charset=US-ASCII 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- Content-Disposition: inline ==============================End of - Headers==============================
First of all I would like to thank everyone for the helpful advice I received earlier on the vapor fixation. The processing of bacteria with paraformaldehyde (3hours) following with 1h of osmium vapor (I tried this combination in reverse as well); vapor dehydration gave me a good fixation as far as I can see at my magnification. Unfortunately, I encountered another problem: salt crystals. By skipping wash I left buffer precipitates seemingly intact yet washing is removing not only salt but also most of the bacteria from the surface treated with antimicrobial. I would greatly appreciate your advice. Albina
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 3, 21 -- From amich-at-ufl.edu Mon Mar 27 11:58:14 2006 3, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 3, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2RHwD3v013670 3, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 11:58:14 -0600 3, 21 -- Received: from osgjas01.cns.ufl.edu (osgjas01.cns.ufl.edu [128.227.74.131]) 3, 21 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k2RHw8kE946362 3, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 12:58:08 -0500 3, 21 -- Message-ID: {608177447.7451143482288345.JavaMail.osg-at-osgjas01.cns.ufl.edu} 3, 21 -- Date: Mon, 27 Mar 2006 12:58:08 -0500 (EST) 3, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 3, 21 -- To: Microscopy-at-microscopy.com 3, 21 -- Subject: Part 2: vapor fixation for SEM 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 3, 21 -- X-Originating-IP: 70.152.34.47 [70.152.34.47] 3, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Hi Albina we have a number of researchers who use vapour fixation for protists using a protocol from Brian Leander's lab. 4% osmium onto a filter paper in the lid of a petri dish and the living sample in the dish base for 30 minutes. Then one drop of 4% osmium added to the mix per ml of liquid. Leave for 30 minutes and dehydrate as normal after putting the specimens onto a nuclearpore filter in a Millipore Swinnex holder.
The vapor fixation results are superb and several euglenoids from Brian Leander have become magazine cover shots.
We have been trying to use the microwave for the dehydration but have found that the samples are not as sticky as when glutaraldehyde is used first and the samples tended to lift off the nucleopore filter in HMDS. This seems to be less of a problem when we use the critical point dryer. However, we have found that if it works conventionally, it will generally work in the microwave at a fraction of the time if you find the right conditions. This is still a work in progress. It seems that some sample is lost even in the critical point dryer. By leaving the filter in the syringe/swinnex holder all the way through the processing, less is lost even with HMDS in the microwave.
Elaine
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- 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============================== 9, 30 -- From ech-at-interchange.ubc.ca Mon Mar 27 13:40:47 2006 9, 30 -- Received: from mta6.mail-relay.ubc.ca (mta6.mail-relay.ubc.ca [137.82.45.12]) 9, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2RJekFT024817 9, 30 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 13:40:47 -0600 9, 30 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 9, 30 -- by mta6.mail-relay.ubc.ca (8.12.11/8.12.11) with ESMTP id k2RJejfl015773 9, 30 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 11:40:45 -0800 (PST) 9, 30 -- (envelope-from ech-at-interchange.ubc.ca) 9, 30 -- Received: from [24.82.105.155] 9, 30 -- (S01060040f4371335.vn.shawcable.net [24.82.105.155]) 9, 30 -- by smtp.interchange.ubc.ca 9, 30 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 9, 30 -- with ESMTPA id {0IWS00MQZXBW5K-at-smtp.interchange.ubc.ca} for 9, 30 -- microscopy-at-microscopy.com; Mon, 27 Mar 2006 11:40:45 -0800 (PST) 9, 30 -- Date: Mon, 27 Mar 2006 11:40:42 -0800 9, 30 -- From: Elaine Humphrey {ech-at-interchange.ubc.ca} 9, 30 -- Subject: Re: [Microscopy] Part 2: vapor fixation for SEM 9, 30 -- In-reply-to: {200603271800.k2RI0BwK016494-at-ns.microscopy.com} 9, 30 -- X-Sender: ech-at-mail.interchange.ubc.ca 9, 30 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 9, 30 -- Cc: amich-at-ufl.edu 9, 30 -- Message-id: {a06100502c04de6b97f2a-at-[24.82.105.155]} 9, 30 -- MIME-version: 1.0 9, 30 -- Content-type: text/plain; format=flowed; charset=us-ascii 9, 30 -- References: {200603271800.k2RI0BwK016494-at-ns.microscopy.com} 9, 30 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.03.27.105106 9, 30 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 9, 30 -- X-PerlMx-Spam: Probability=7%, Report=IP_HTTP_ADDR 0, __C230066_P1_5 0, __C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0 9, 30 -- X-Spam-Level: 9, 30 -- X-Spam-Flag: No ==============================End of - Headers==============================
thank you for the advice. I will look up Brian Leander's publications. My primary objective is to evaluate changes in the bacteria inhibited by antimicrobial agents. The challenge is working with fibrous substrate inoculated with bacteria to test antimicrobial treatments. In many cases I would like avoid wetting substrates; so vapor fixation is a way to go with the exception of salt residue left behind. Even minimal wash removes bacteria from the treated substrate because they adhesive properties are compromised yet without wash salt crystals are obscuring view. Albina
On Mon Mar 27 14:40:42 EST 2006, Elaine Humphrey {ech-at-interchange.ubc.ca} wrote:
} Hi Albina } we have a number of researchers who use vapour fixation for } protists using a protocol from Brian Leander's lab. 4% osmium } onto a filter paper in the lid of a petri dish and the living } sample in the dish base for 30 minutes. Then one drop of 4% } osmium added to the mix per ml of liquid. Leave for 30 minutes } and dehydrate as normal after putting the specimens onto a } nuclearpore filter in a Millipore Swinnex holder. } } The vapor fixation results are superb and several euglenoids from } Brian Leander have become magazine cover shots. } } We have been trying to use the microwave for the dehydration but } have found that the samples are not as sticky as when } glutaraldehyde is used first and the samples tended to lift off } the nucleopore filter in HMDS. This seems to be less of a problem } when we use the critical point dryer. However, we have found that } if it works conventionally, it will generally work in the } microwave at a fraction of the time if you find the right } conditions. This is still a work in progress. It seems that some } sample is lost even in the critical point dryer. By leaving the } filter in the syringe/swinnex holder all the way through the } processing, less is lost even with HMDS in the microwave. } } Elaine } } } } } ---------------------------------------------------------------------------- } } 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 } } ---------------------------------------------------------------------------- } } } } First of all I would like to thank everyone for the helpful } } advice } } I received earlier on the vapor fixation. The processing of } } bacteria with paraformaldehyde (3hours) following with 1h of } } osmium vapor (I tried this combination in reverse as well); vapor } } dehydration gave me a good fixation as far as I can see at my } } magnification. } } Unfortunately, I encountered another problem: salt crystals. By } } skipping wash I left buffer precipitates seemingly intact yet } } washing is removing not only salt but also most of the bacteria } } from the surface treated with antimicrobial. I would greatly } } appreciate your advice. } } Albina } } } } -- } } MIKHAYLOVA,ALBINA, PhD } } Post Doctoral Research Associate } } Materials Science and Engineering } } University of Florida } } } } } } ==============================Original } } Headers============================== } } 3, 21 -- From amich-at-ufl.edu Mon Mar 27 11:58:14 2006 } } 3, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu } } [128.227.74.165]) } } 3, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k2RHwD3v013670 } } 3, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 } } 11:58:14 -0600 } } 3, 21 -- Received: from osgjas01.cns.ufl.edu } } (osgjas01.cns.ufl.edu [128.227.74.131]) } } 3, 21 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id } } k2RHw8kE946362 } } 3, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 } } 12:58:08 -0500 } } 3, 21 -- Message-ID: } } {608177447.7451143482288345.JavaMail.osg-at-osgjas01.cns.ufl.edu} } } 3, 21 -- Date: Mon, 27 Mar 2006 12:58:08 -0500 (EST) } } 3, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} } } 3, 21 -- To: Microscopy-at-microscopy.com } } 3, 21 -- Subject: Part 2: vapor fixation for SEM } } 3, 21 -- MIME-Version: 1.0 } } 3, 21 -- Content-Type: text/plain; format=flowed; } } charset=us-ascii } } 3, 21 -- Content-Transfer-Encoding: 7bit } } 3, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) } } 3, 21 -- X-Originating-IP: 70.152.34.47 [70.152.34.47] } } 3, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED } } 3, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, } } tests=ALL_TRUSTED } } 3, 21 -- X-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } 3, 21 -- X-UFL-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } ==============================End of - } } Headers============================== } } } -- 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 } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 8, 22 -- From amich-at-ufl.edu Mon Mar 27 13:57:56 2006 8, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2RJvsxx002025 8, 22 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 13:57:55 -0600 8, 22 -- Received: from osgjas01.cns.ufl.edu (osgjas01.cns.ufl.edu [128.227.74.131]) 8, 22 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k2RJvlVY3825720; 8, 22 -- Mon, 27 Mar 2006 14:57:47 -0500 8, 22 -- Message-ID: {2095729987.15611143489467440.JavaMail.osg-at-osgjas01.cns.ufl.edu} 8, 22 -- Date: Mon, 27 Mar 2006 14:57:47 -0500 (EST) 8, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 8, 22 -- To: Elaine Humphrey {ech-at-interchange.ubc.ca} , 8, 22 -- Microscopy Listserver {microscopy-at-microscopy.com} 8, 22 -- Subject: Re: [Microscopy] Part 2: vapor fixation for SEM 8, 22 -- MIME-Version: 1.0 8, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 8, 22 -- Content-Transfer-Encoding: 7bit 8, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 8, 22 -- X-Originating-IP: 70.152.34.47 [70.152.34.47] 8, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 8, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Dear Basil, Many years ago , when we first started to try to do cross-sections, we used Devcon 2-Ton epoxy for the cross-sections, while we were waiting for the M-610 Bond to arrive. It is not as thin as the M-610, but with pressure on our home-built parallel-jaw clamp the glue joints were thin enough and we got good results. This is a slow, 24 hour cure epoxy. Good luck, 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: {bjulies-at-uwc.ac.za} To: {mager-at-interchange.ubc.ca} Sent: Monday, March 27, 2006 7:26 AM
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Email: isabeln-at-popsrv.ist.utl.pt Name: Isabel Dias Nogueira
Organization: MicroLab - Instituto Superior Tecnico
Title-Subject: [Filtered] FEG-SEM: cold cathode versus Schottky
Question: Our lab is in the process of buying a FEG-SEM. The main question at this point is whether to choose cold cathode (higher resolution) or Schottky emission (higher current). At first we thought we should go for cold cathode because of the resolution, but we also plan to acquire a EBSD (diffraction using Kikuchi lines) and a EDS detector for mapping, both requiring high beam current. The EBSD, in particular, also requires long acquisition times, which may not work well with the need of flashing the cold cathode emitter every 10h or so. Another issue is current stability: will the lower current stability of the cold cathode (5%) have any consequences on quantifying point EDS analysis ? I would appreciate any comments from users of both types of FEG-SEM, and also of EBSD and EDS mapping. Thanks,
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Email: p.arico-at-email.it Name: Pietro Arico'
Organization: Dept. CFTA - University of Palermo (IT)
Title-Subject: [Filtered] EDS Co Calibration
Question: dear all, i am trying to acquire spectra of silicate and metallic standards using a LEO 440 coupled with an Oxford Link Isis 300 EDS system. I have acquired 37 spectra of different standards doing a calibration using Cobalt to check the instrument stability (I have no Faraday cup and Current meter). Here are the values of the calibrations: Time 14.15 14.25 14.35 14.45 14.55 15.05 15.35 16.15 16.35 16.55 17.15 Zero energy channel 9.623245 9.622585 9.619363 9.624291 9.623804 9.626814 9.623498 9.623422 9.623599 9.624767 9.624976 Energy per channel (eV) 20.00182 19.99826 20.00041 20.00109 19.99679 19.99775 19.99981 19.99792 20.00136 20.00056 19.99934 Counts in calibration peak 48880 48736 48457 48225 47876 48331 49103 48991 49426 49188 48925
do you think the instrument is enough stable or not? the differences between these values are negligible or not? thank you very much fopr your help (it's the first time I try to do this!!!)
Pietro Arico' Dept. Chemistry and Physics of the Earth (CFTA) University of Palermo Via Archirafi, 36 Palermo, 90123 - Italy email: p.arico-at-email.it
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Email: ycn1-at-psu.edu Name: Yuk-Chow Ng
Organization: Penn State University
Title-Subject: [Filtered] quantitation of immunofluorescence
Question: I am trying to perform a semi-quantitative comparison of immunofluorescence intensity on the sarcoplasmic membrane of skeletal muscle fibers (cross sections). Is there a way to trace the signals on the membrane of multiple fibers and compare the overall intensity between a control and a treated sample (sections).
The calibration of EDS is usually done at Al and Cu.
I use X-Checker Extra to do this since it does Al and Cu plus F, Be, C, B, N.
The first pass is Al+Cu. Then separate passes for lower Z elements are lower KV.
Disclaimer: I have no financial interest in X-checker.
gary g.
At 04:55 PM 3/27/2006, you wrote:
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==============================Original Headers============================== 12, 20 -- From gary-at-gaugler.com Mon Mar 27 19:16:55 2006 12, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S1GqRr022289 12, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 19:16:53 -0600 12, 20 -- Received: (qmail 12667 invoked from network); 27 Mar 2006 17:16:50 -0800 12, 20 -- Received: by simscan 1.1.0 ppid: 12664, pid: 12665, t: 0.1680s 12, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 20 -- by qsmtp3 with SMTP; 27 Mar 2006 17:16:50 -0800 12, 20 -- Message-Id: {7.0.1.0.2.20060327171410.026a6928-at-gaugler.com} 12, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 20 -- Date: Mon, 27 Mar 2006 17:16:51 -0800 12, 20 -- To: p.arico-at-email.it 12, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: EDS Co Calibration 12, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 20 -- In-Reply-To: {200603280055.k2S0tCFc003300-at-ns.microscopy.com} 12, 20 -- References: {200603280055.k2S0tCFc003300-at-ns.microscopy.com} 12, 20 -- Mime-Version: 1.0 12, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Well, this is a significant decision point indeed.
I don't necessarily agree that cold cathode FE SEMs produce higher resolution. But that is not the issue here.
Schottky is going to produce a very high current beam with astounding stability. This is nice but critical for EBSD. However, what is the maximum probe current available? You can have great stability of a low current system and spend hours on an EBSD scan. You also have to figure out which EBSD system you are going to use. TSL offers drift correction (good at } 1KX) while AFIK, HKL does not offer. These are your main two providers of EBSD.
Your beam strength will greatly impact the fps of the EBSD data collection. fps of between 22-33 are good. Higher fps may be dependent on degradation of probe diameter. So watch out for this.
Now, throwing in EDS mapping you are moving another step. These maps can take hours to complete--depending on cps and DT. Either way, the Schottky FE is going to be superior, IMO.
Bottom line...get a Schottky FE with as high a probe current as you can get with adjustable probe diameters. BTW, each SEM maker does this differently. Most use final apertures. Zeiss does not.
Contact me off-line if you would like some specifics.
Disclaimer: No financial interest in any supplier I reference.
gary g.
At 04:55 PM 3/27/2006, you wrote:
} 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 isabeln-at-popsrv.ist.utl.pt as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: isabeln-at-popsrv.ist.utl.pt } Name: Isabel Dias Nogueira } } Organization: MicroLab - Instituto Superior Tecnico } } Title-Subject: [Filtered] FEG-SEM: cold cathode versus Schottky } } Question: Our lab is in the process of buying a FEG-SEM. } The main question at this point is whether to choose cold cathode } (higher resolution) or Schottky emission (higher current). At first } we thought we should go for cold cathode because of the resolution, } but we also plan to acquire a EBSD (diffraction using Kikuchi lines) } and a EDS detector for mapping, both requiring high beam current. } The EBSD, in particular, also requires long acquisition times, which } may not work well with the need of flashing the cold cathode emitter } every 10h or so. Another issue is current stability: will the lower } current stability of the cold cathode (5%) have any consequences on } quantifying point EDS analysis ? I would appreciate any comments } from users of both types of FEG-SEM, and also of EBSD and EDS mapping. Thanks, } } Isabel } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 7, 12 -- From zaluzec-at-microscopy.com Mon Mar 27 18:52:08 2006 } 7, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 7, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k2S0q8n0026146 } 7, 12 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 } 18:52:08 -0600 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: (Unverified) } 7, 12 -- Message-Id: {p06110407c04e3929fcf5-at-[206.69.208.22]} } 7, 12 -- Date: Mon, 27 Mar 2006 18:52:06 -0600 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: isabeln-at-mail.ist.utl.pt (by way of MicroscopyListserver) } 7, 12 -- Subject: viaWWW: FEG-SEM: cold cathode versus Schottky } 7, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
==============================Original Headers============================== 14, 20 -- From gary-at-gaugler.com Mon Mar 27 19:29:38 2006 14, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S1Tceo031943 14, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 19:29:38 -0600 14, 20 -- Received: (qmail 987 invoked from network); 27 Mar 2006 17:28:55 -0800 14, 20 -- Received: by simscan 1.1.0 ppid: 981, pid: 983, t: 0.0845s 14, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 14, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 14, 20 -- by qsmtp4 with SMTP; 27 Mar 2006 17:28:54 -0800 14, 20 -- Message-Id: {7.0.1.0.2.20060327171705.026b7008-at-gaugler.com} 14, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 20 -- Date: Mon, 27 Mar 2006 17:29:38 -0800 14, 20 -- To: isabeln-at-mail.ist.utl.pt 14, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 14, 20 -- Subject: Re: [Microscopy] viaWWW: FEG-SEM: cold cathode versus Schottky 14, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 14, 20 -- In-Reply-To: {200603280055.k2S0tCiJ003290-at-ns.microscopy.com} 14, 20 -- References: {200603280055.k2S0tCiJ003290-at-ns.microscopy.com} 14, 20 -- Mime-Version: 1.0 14, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
First of all be careful when taking about signal intensity in fluorescence, especially immunofluorescence, since the amount of signal is not directly related to the concentration of the target. You may however compare the intensity of 2 samples treated exactly the same way, still being careful not to draw too precise conclusions. Microscopy is not the method of choice for quantification. For your special need, I suppose that using a confocal microscope you could draw a profile of fluorescence or define a ROI (region of interest) and let the computer calculate the amount of fluorescence. Don't forget to use the same settings for all samples otherwise no comparison is possible! (manipulation of signal is so easy on a confocal!)
Stephane-without-i
--- ycn1-at-psu.edu wrote:
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Tue Mar 28 00:31:16 2006 9, 20 -- Received: from web37401.mail.mud.yahoo.com (web37401.mail.mud.yahoo.com [209.191.87.54]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S6VG8g012752 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 00:31:16 -0600 9, 20 -- Received: (qmail 24141 invoked by uid 60001); 28 Mar 2006 06:31:15 -0000 9, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 20 -- s=s1024; d=yahoo.com; 9, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 20 -- b=nuWJ184L0ZuIRfCk4NQVTL/5DM1hFCHLEt4345+F3T0KpaNJGXrKBvp0I7X3zFAmIr0Y8o7NqY/GrcEt7FaqEOONq999+UYTHM/8zgtmpCE++HkveAay0WwToTVDeuk/Yvm6u7o22XeU/I1VOiOwOg3x2cjFwH127a2GqqcQ7Os= ; 9, 20 -- Message-ID: {20060328063115.24139.qmail-at-web37401.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37401.mail.mud.yahoo.com via HTTP; Mon, 27 Mar 2006 22:31:15 PST 9, 20 -- Date: Mon, 27 Mar 2006 22:31:15 -0800 (PST) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] viaWWW: quantitation of immunofluorescence 9, 20 -- To: ycn1-at-psu.edu 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200603280101.k2S11SjV020852-at-ns.microscopy.com} 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; charset=iso-8859-1 9, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I have never made a detailed study about superglue outgassing, but when used as a crevice and pore back-fill on a polished specimens (and suitably cured) I have not noticed a vacuum problem. This is in the area of 5E-5 to 5E-6 Torr. This is not to say it does not outgas, but only that my pumping system has no trouble keeping up.
OTOH, it is NOT beam stable. I have seen it boil under the beam at anything less than very low kV and current.
Regards, Woody White BWXT Services
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Dear Colleagues,
I would appreciate it if some of you could give me your experience of how the widely available Superglue behaves under vacuum in the SEM and TEM.
Is it a suitable adhesive?
Is there too much de-gassing etc.
Many thanks,
Ted Dunn The EMscope Company Ltd.
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==============================Original Headers============================== 10, 19 -- From drteddunne-at-yahoo.com Tue Mar 28 03:45:05 2006 10, 19 -- Received: from web33407.mail.mud.yahoo.com (web33407.mail.mud.yahoo.com [68.142.206.139]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S9j4Qn026844 10, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 03:45:04 -0600 10, 19 -- Received: (qmail 73056 invoked by uid 60001); 28 Mar 2006 09:45:04 -0000 10, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 19 -- s=s1024; d=yahoo.com; 10, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Cont ent-Type:Content-Transfer-Encoding; 10, 19 -- b=qc0c3Nz3PGKgGIW9L3FWLolqeL2G4o9CtPkFC+5Zckimur+GWWIvWx9YYEy7qpRMWlAQAq QAbcOIj7cNINrfdTJti374bhqp+etRhqMY3hXdHKg7AaggEpXQCX3TvlXm+kRRetbpQOK3ix qg58Ocryz8g0IcMkqPmgjr/RKr9JQ= ; 10, 19 -- Message-ID: {20060328094504.73054.qmail-at-web33407.mail.mud.yahoo.com} 10, 19 -- Received: from [202.47.247.116] by web33407.mail.mud.yahoo.com via HTTP; Tue, 28 Mar 2006 01:45:04 PST 10, 19 -- Date: Tue, 28 Mar 2006 01:45:04 -0800 (PST) 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 19 -- Subject: Superglue 10, 19 -- To: microscopy-at-microscopy.com 10, 19 -- In-Reply-To: {200603272125.k2RLPJn3019014-at-ns.microscopy.com} 10, 19 -- MIME-Version: 1.0 10, 19 -- Content-Type: text/plain; charset=iso-8859-1 10, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 19, 27 -- From nwwhite-at-bwxt.com Tue Mar 28 07:04:34 2006 19, 27 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 19, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SD4XQC016566 19, 27 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Mar 2006 07:04:33 -0600 19, 27 -- Received: from ([131.184.13.224]) 19, 27 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.545394; 19, 27 -- Tue, 28 Mar 2006 08:04:20 -0500 19, 27 -- Received: from bwxslynpo01.BWXS.BWXTECH.NET ([131.184.13.4]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 19, 27 -- Tue, 28 Mar 2006 08:04:19 -0500 19, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 19, 27 -- Content-class: urn:content-classes:message 19, 27 -- MIME-Version: 1.0 19, 27 -- Content-Type: text/plain; 19, 27 -- charset="US-ASCII" 19, 27 -- Subject: RE: [Microscopy] Superglue 19, 27 -- Date: Tue, 28 Mar 2006 08:04:19 -0500 19, 27 -- Message-ID: {70E4EA352EC987489773D55B0FF83E1C1DF9C4-at-bwxslynpo01.BWXS.BWXTECH.NET} 19, 27 -- X-MS-Has-Attach: 19, 27 -- X-MS-TNEF-Correlator: 19, 27 -- Thread-Topic: [Microscopy] Superglue 19, 27 -- Thread-Index: AcZSTH+0uE/ojL+RRHKVxLWBujyaFgAGnfog 19, 27 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 19, 27 -- To: {drteddunne-at-yahoo.com} , 19, 27 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 19, 27 -- X-OriginalArrivalTime: 28 Mar 2006 13:04:19.0972 (UTC) FILETIME=[205D9C40:01C65268] 19, 27 -- Content-Transfer-Encoding: 8bit 19, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2SD4XQC016566 ==============================End of - Headers==============================
I am posting message for Dr. Milos Kalab who is not a subscriber. His web pages have been recommended from time to time on the list.
His messages are as following: Milos Kalab, Honorary Research Associate at Agriculture and Agri-Food Canada in Ottawa, who has many websites on electron microscopy of foods and microorganisms hosted by the generosity of the University of Lund in Sweden on their server with URL either http://distans.livstek.lth.se:2080/ or http://anka.livstek.lth.se:2080/ wants to apologize to their visitors. The server in Sweden has been out of order and it is not known when it will be repaired. The starting point with links to those websites is in Canada at http://www.magma.ca/~scimat/ where new information may be found. Milos has been transferring some of the websites to active addresses to make them accessible again. He may be contacted at scimat-at-magma.ca . Thank you.
Ann Fook Yang Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 Room 2097, K.W. Neatby Bldg., CEF , Ottawa, ON, Canada K1A 0C6 yanga-at-agr.gc.ca
==============================Original Headers============================== 3, 26 -- From YANGA-at-AGR.GC.CA Tue Mar 28 09:22:55 2006 3, 26 -- Received: from agrpazsmtp2.agr.gc.ca (agrpazsmtp2.agr.gc.ca [192.197.71.137]) 3, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFMtF3031662 3, 26 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 09:22:55 -0600 3, 26 -- Received: from onncrxcn1.AGR.GC.CA ([192.168.122.1]) 3, 26 -- by agrpazsmtp2.agr.gc.ca (8.13.6/8.13.3) with ESMTP id k2SFJ0mu023222 3, 26 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:19:00 -0500 3, 26 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn1.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 3, 26 -- Tue, 28 Mar 2006 10:22:55 -0500 3, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 3, 26 -- content-class: urn:content-classes:message 3, 26 -- MIME-Version: 1.0 3, 26 -- Content-Type: text/plain; 3, 26 -- charset="iso-8859-1" 3, 26 -- Subject: web problem 3, 26 -- Date: Tue, 28 Mar 2006 10:22:54 -0500 3, 26 -- Message-ID: {E035A9C87303AE4AB9BA10FD8324DFB1024334B6-at-onncrxms3.agr.gc.ca} 3, 26 -- X-MS-Has-Attach: 3, 26 -- X-MS-TNEF-Correlator: 3, 26 -- Thread-Topic: web problem 3, 26 -- Thread-Index: AcZR2hZo/1G+jdJlQ9e2aCztWikm7Q== 3, 26 -- From: "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 3, 26 -- To: {microscopy-at-microscopy.com} 3, 26 -- X-OriginalArrivalTime: 28 Mar 2006 15:22:55.0132 (UTC) FILETIME=[7C9655C0:01C6527B] 3, 26 -- Content-Transfer-Encoding: 8bit 3, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2SFMtF3031662 ==============================End of - Headers==============================
Just a short note to thanks everyone who e-mailed or called me to provide assistance with the identification of titanium from titanium oxide in an organic binder. Your assistance was much appreciated.
If your interested, I ashed my sample to reduce the organic fraction and fluxed the remaining material in sodium borate with a platinum wire and alcohol lamp (How many labs still have alcohol lamps…). The bead was removed, crushed and dissolved in concentrated H2SO4. Both the hydrogen peroxide and chromotropic acid tests as described as per Feigl in “Qualitative Analysis of Spot Tests†were used. I tried both tests as a spot test on filter paper, in a capillary tube and in a white spot plate. The spot test worked best for me.
Simply heating a sample of TiO2 in concentrated H2SO4 did not convert much of the relatively inert TiO2 into a detectable form, but fluxing did.
Thanks again!!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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==============================Original Headers============================== 10, 19 -- From frank.karl-at-degussa.com Tue Mar 28 09:38:42 2006 10, 19 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFcgoh008854 10, 19 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Mar 2006 09:38:42 -0600 10, 19 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 10, 19 -- by framailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2SFcdoH024564 10, 19 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Mar 2006 17:38:40 +0200 10, 19 -- Subject: Yes, my sample contained titanium 10, 19 -- To: microscopy-at-msa.microscopy.com 10, 19 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 10, 19 -- Message-ID: {OF668E6ACA.056DB750-ON8525713F.0055C11D-8525713F.0055EC98-at-degussa.com} 10, 19 -- From: frank.karl-at-degussa.com 10, 19 -- Date: Tue, 28 Mar 2006 10:38:31 -0500 10, 19 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 10, 19 -- 03/28/2006 09:38:41 AM 10, 19 -- MIME-Version: 1.0 10, 19 -- Content-type: text/plain; charset=UTF-8 10, 19 -- Content-Transfer-Encoding: 8bit 10, 19 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k2SFcgoh008854 ==============================End of - Headers==============================
I use Gatan's G-1 epoxy for all my cross sections. (You can get the same product from Epo-tek as well, I believe they are the original manufacturer). I have not tried it with Ti substrates, but I routinely do silicon, glass, MgO, SrTiO3, etc. I wouldn't call it low viscosity, but I regularly get glue lines less than 10 nm as measured in the TEM. I cure it on a hotplate around 100C in a vice with as much pressure as I can get. Hope that helps!
Leslie
Leslie Krupp (Thompson) Engineer/Scientiest IBM Almaden Research 650 Harry Road, K19/D2 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 4, 27 -- From lkrupp-at-us.ibm.com Tue Mar 28 09:44:33 2006 4, 27 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) 4, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFiX1O018381 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 09:44:33 -0600 4, 27 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 4, 27 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k2SFiWIs016723 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:33 -0500 4, 27 -- Received: from d01av01.pok.ibm.com (d01av01.pok.ibm.com [9.56.224.215]) 4, 27 -- by d01relay04.pok.ibm.com (8.12.10/NCO/VER6.8) with ESMTP id k2SFiMFj176842 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:22 -0500 4, 27 -- Received: from d01av01.pok.ibm.com (loopback [127.0.0.1]) 4, 27 -- by d01av01.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k2SFiMvs012415 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:22 -0500 4, 27 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 4, 27 -- by d01av01.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k2SFiM2N012390 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:22 -0500 4, 27 -- To: microscopy-at-microscopy.com 4, 27 -- MIME-Version: 1.0 4, 27 -- Subject: Re: Adehsive for Titanium 4, 27 -- X-Mailer: Lotus Notes Release 6.0.2CF1 June 9, 2003 4, 27 -- Message-ID: {OF017C7D97.579B9454-ON8525713F.0055E41F-8825713F.00567582-at-us.ibm.com} 4, 27 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 4, 27 -- Date: Tue, 28 Mar 2006 07:44:21 -0800 4, 27 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF18 | February 28, 2006) at 4, 27 -- 03/28/2006 10:44:22, 4, 27 -- Serialize complete at 03/28/2006 10:44:22 4, 27 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
While outgassing may not be a problem I have frequently had issues with bond failure when exposed to temperature excursions. Thus I do not use superglue for anything I want permanently bonded.
Much better are the epoxies, such as Gatan's G-1 or the M-Bond brand. These are stable under the e-beam and do not outgas.
-- Roger A. Ristau, PhD TEM Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269 vox: 860-486-5379 fax: 860-486-4745
==============================Original Headers============================== 6, 19 -- From raristau-at-ims.uconn.edu Tue Mar 28 09:54:58 2006 6, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFsvkG027967 6, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 09:54:58 -0600 6, 19 -- Received: from [137.99.44.251] (d44h251.public.uconn.edu [137.99.44.251]) 6, 19 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k2SFsZXC022431 6, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:54:35 -0500 6, 19 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 19 -- Date: Tue, 28 Mar 2006 10:54:18 -0500 6, 19 -- Subject: re: Superglue 6, 19 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 6, 19 -- To: {microscopy-at-microscopy.com} 6, 19 -- Message-ID: {C04EC65A.AD8%raristau-at-ims.uconn.edu} 6, 19 -- Mime-version: 1.0 6, 19 -- Content-type: text/plain; charset="US-ASCII" 6, 19 -- Content-transfer-encoding: 7bit 6, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 6, 19 -- X-UConn-MailScanner: Found to be clean 6, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
It's not a micro-chemical test, but TAPPI Test Method T627 "Determination of Titanium dioxide" uses ammonium sulfate in sulfuric acid. They also list an XRF test, method T554. Methods can be purchased individually from www.tappi.org.
I have no financial intesest in TAPPI, a paper industry association.
David Rothbard Bureau of Engraving and Printing
frank.karl-at-degussa.com wrote:
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==============================Original Headers============================== 11, 20 -- From rothbardd-at-netscape.net Tue Mar 28 11:38:47 2006 11, 20 -- Received: from imo-d01.mx.aol.com (imo-d01.mx.aol.com [205.188.157.33]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SHckpL006710 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 11:38:46 -0600 11, 20 -- Received: from rothbardd-at-netscape.net 11, 20 -- by imo-d01.mx.aol.com (mail_out_v38_r7.3.) id w.13b.13377e91 (16237) 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 12:38:42 -0500 (EST) 11, 20 -- Received: from netscape.net (mow-d20.webmail.aol.com [205.188.139.161]) by air-in03.mx.aol.com (v108_r3.6) with ESMTP id MAILININ31-3f6d442974a2233; Tue, 28 Mar 2006 12:38:42 -0500 11, 20 -- Date: Tue, 28 Mar 2006 12:38:42 -0500 11, 20 -- From: rothbardd-at-netscape.net 11, 20 -- To: microscopy-at-microscopy.com 11, 20 -- Subject: RE: Question Microchemical test for Ti 11, 20 -- MIME-Version: 1.0 11, 20 -- Message-ID: {38C1CBEB.7C245EF9.034D9A6A-at-netscape.net} 11, 20 -- X-Mailer: Atlas Mailer 2.0 11, 20 -- X-AOL-IP: 199.196.144.13 11, 20 -- X-AOL-Language: english 11, 20 -- Content-Type: text/plain; charset=iso-8859-1 11, 20 -- Content-Transfer-Encoding: 8bit 11, 20 -- X-Spam-Flag: NO ==============================End of - Headers==============================
Ted: I've used "Super Glue" (the Loctite variety, mostly) for years to mount samples for cross-sectioning and then imaging in the SEM, both thermionic and cold FE types. I've had no problems with outgassing, even with my JEOL 6600FXV, which is very sensitive to any outgassing.
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 } ---------------------------------------------------------------------------- } } Dear Colleagues, } } I would appreciate it if some of you could give me } your experience of how the widely available Superglue } behaves under vacuum in the SEM and TEM. } } Is it a suitable adhesive? } } Is there too much de-gassing etc. } } Many thanks, } } } Ted Dunn } The EMscope Company Ltd. } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 10, 19 -- From drteddunne-at-yahoo.com Tue Mar 28 03:45:05 2006 } 10, 19 -- Received: from web33407.mail.mud.yahoo.com (web33407.mail.mud.yahoo.com [68.142.206.139]) } 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S9j4Qn026844 } 10, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 03:45:04 -0600 } 10, 19 -- Received: (qmail 73056 invoked by uid 60001); 28 Mar 2006 09:45:04 -0000 } 10, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 10, 19 -- s=s1024; d=yahoo.com; } 10, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 10, 19 -- b=qc0c3Nz3PGKgGIW9L3FWLolqeL2G4o9CtPkFC+5Zckimur+GWWIvWx9YYEy7qpRMWlAQAqQAbcOIj7cNINrfdTJti374bhqp+etRhqMY3hXdHKg7AaggEpXQCX3TvlXm+kRRetbpQOK3ixqg58Ocryz8g0IcMkqPmgjr/RKr9JQ= ; } 10, 19 -- Message-ID: {20060328094504.73054.qmail-at-web33407.mail.mud.yahoo.com} } 10, 19 -- Received: from [202.47.247.116] by web33407.mail.mud.yahoo.com via HTTP; Tue, 28 Mar 2006 01:45:04 PST } 10, 19 -- Date: Tue, 28 Mar 2006 01:45:04 -0800 (PST) } 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} } 10, 19 -- Subject: Superglue } 10, 19 -- To: microscopy-at-microscopy.com } 10, 19 -- In-Reply-To: {200603272125.k2RLPJn3019014-at-ns.microscopy.com} } 10, 19 -- MIME-Version: 1.0 } 10, 19 -- Content-Type: text/plain; charset=iso-8859-1 } 10, 19 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers============================== } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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============================== 4, 22 -- From r-holdford-at-ti.com Tue Mar 28 14:01:56 2006 4, 22 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SK1tsJ018441 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 28 Mar 2006 14:01:55 -0600 4, 22 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 22 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k2SK1jHk027781; 4, 22 -- Tue, 28 Mar 2006 14:01:55 -0600 (CST) 4, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 22 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k2SK1jqq026509; 4, 22 -- Tue, 28 Mar 2006 14:01:45 -0600 (CST) 4, 22 -- Message-ID: {44299628.5050003-at-ti.com} 4, 22 -- Date: Tue, 28 Mar 2006 14:01:44 -0600 4, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 22 -- Organization: SC Packaging Development -- FA Development 4, 22 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 4, 22 -- MIME-Version: 1.0 4, 22 -- To: drteddunne-at-yahoo.com, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Re: [Microscopy] Superglue 4, 22 -- References: {200603280945.k2S9jRoG027186-at-ns.microscopy.com} 4, 22 -- In-Reply-To: {200603280945.k2S9jRoG027186-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I've also used Loctite 460 to attach samples to a copper grid for the TEM (using the least amount of glue possible). Prior to putting in the TEM, I cure the glue in our lab vacuum desiccator for ~1 hour. Again, never had a problem.
-----Original Message----- X-from: drteddunne-at-yahoo.com [mailto:drteddunne-at-yahoo.com] Sent: Tuesday, March 28, 2006 3:51 AM To: wgstratton-at-wisc.edu
Dear Colleagues,
I would appreciate it if some of you could give me your experience of how the widely available Superglue behaves under vacuum in the SEM and TEM.
Is it a suitable adhesive?
Is there too much de-gassing etc.
Many thanks,
Ted Dunn The EMscope Company Ltd.
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==============================Original Headers============================== 10, 19 -- From drteddunne-at-yahoo.com Tue Mar 28 03:45:05 2006 10, 19 -- Received: from web33407.mail.mud.yahoo.com (web33407.mail.mud.yahoo.com [68.142.206.139]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S9j4Qn026844 10, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 03:45:04 -0600 10, 19 -- Received: (qmail 73056 invoked by uid 60001); 28 Mar 2006 09:45:04 -0000 10, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 19 -- s=s1024; d=yahoo.com; 10, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Cont ent-Type:Content-Transfer-Encoding; 10, 19 -- b=qc0c3Nz3PGKgGIW9L3FWLolqeL2G4o9CtPkFC+5Zckimur+GWWIvWx9YYEy7qpRMWlAQAq QAbcOIj7cNINrfdTJti374bhqp+etRhqMY3hXdHKg7AaggEpXQCX3TvlXm+kRRetbpQOK3ix qg58Ocryz8g0IcMkqPmgjr/RKr9JQ= ; 10, 19 -- Message-ID: {20060328094504.73054.qmail-at-web33407.mail.mud.yahoo.com} 10, 19 -- Received: from [202.47.247.116] by web33407.mail.mud.yahoo.com via HTTP; Tue, 28 Mar 2006 01:45:04 PST 10, 19 -- Date: Tue, 28 Mar 2006 01:45:04 -0800 (PST) 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 19 -- Subject: Superglue 10, 19 -- To: microscopy-at-microscopy.com 10, 19 -- In-Reply-To: {200603272125.k2RLPJn3019014-at-ns.microscopy.com} 10, 19 -- MIME-Version: 1.0 10, 19 -- Content-Type: text/plain; charset=iso-8859-1 10, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 18, 24 -- From wgstratton-at-wisc.edu Tue Mar 28 14:31:54 2006 18, 24 -- Received: from mail.cae.wisc.edu (mail.cae.wisc.edu [144.92.13.31]) 18, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SKVsak028699 18, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 14:31:54 -0600 18, 24 -- Received: from trogdor (trogdor.msae.wisc.edu [128.104.200.76]) 18, 24 -- by mail.cae.wisc.edu (8.13.3+Sun/8.13.4) with ESMTP id k2SKVqEM009719; 18, 24 -- Tue, 28 Mar 2006 14:31:52 -0600 (CST) 18, 24 -- From: "William Stratton" {wgstratton-at-wisc.edu} 18, 24 -- To: {drteddunne-at-yahoo.com} , {Microscopy-at-microscopy.com} 18, 24 -- Subject: RE: [Microscopy] Superglue 18, 24 -- Date: Tue, 28 Mar 2006 14:31:46 -0600 18, 24 -- Message-ID: {000c01c652a6$a748c250$4cc86880-at-trogdor} 18, 24 -- MIME-Version: 1.0 18, 24 -- Content-Type: text/plain; 18, 24 -- charset="us-ascii" 18, 24 -- Content-Transfer-Encoding: 7bit 18, 24 -- X-Priority: 3 (Normal) 18, 24 -- X-MSMail-Priority: Normal 18, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 18, 24 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2527 18, 24 -- Importance: Normal 18, 24 -- In-Reply-To: {200603280950.k2S9oYIc031851-at-ns.microscopy.com} 18, 24 -- X-CAE-MailScanner-Information: Please contact security-at-engr.wisc.edu if this message contains a virus or has been corrupted in delivery. 18, 24 -- X-CAE-MailScanner: Found to be clean (benji) ==============================End of - Headers==============================
yes, I used G-1 epoxy from Gatan. I usually leave it in air for about 10minutes before heating/curing the epoxy in oven, so I can have very thin glue line.
I am pretty sure that G-1 epoxy works for Ti. I have prapared some bio-samples grown on Ti-substrates. It worked very well. My colleague also used the G-1 epoxy to prepare the samples (they put samples in a Ti-clip supplied by a Hungary comapny, I can nor remember the name), it works well.
---- Original message ---- } Date: Tue, 28 Mar 2006 09:45:25 -0600 } From: lkrupp-at-us.ibm.com } Subject: [Microscopy] Re: Adehsive for Titanium } To: clei-at-uiuc.edu
} Basil- } } I use Gatan's G-1 epoxy for all my cross sections. (You can get the same } product from Epo-tek as well, I believe they are the original } manufacturer). I have not tried it with Ti substrates, but I routinely do } silicon, glass, MgO, SrTiO3, etc. I wouldn't call it low viscosity, but I } regularly get glue lines less than 10 nm as measured in the TEM. I cure } it on a hotplate around 100C in a vice with as much pressure as I can get. } Hope that helps! } } Leslie } } Leslie Krupp (Thompson) } Engineer/Scientiest } IBM Almaden Research } 650 Harry Road, K19/D2 } San Jose, CA 95120-6099 } (408) 927-3856 } } ==============================Original Changhui LEI ******************************* Research Electron Microscopist Center for Microanalysis of Materials Frederick Seitz Materials Research Laboratory 104 S. Goodwin Avenue Urbana, IL 61801 USA email: clei-at-uiuc.edu tel: 1-217-244-6177 fax: 1-217-244-2178
==============================Original Headers============================== 6, 26 -- From clei-at-uiuc.edu Tue Mar 28 15:52:40 2006 6, 26 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu [128.174.5.96]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SLqeOK006831 6, 26 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 15:52:40 -0600 6, 26 -- Received: from expms6.cites.uiuc.edu (expms6.cites.uiuc.edu [128.174.5.43]) 6, 26 -- by expredir5.cites.uiuc.edu (8.13.6/8.13.6) with ESMTP id k2SLqdVP013360; 6, 26 -- Tue, 28 Mar 2006 15:52:39 -0600 (CST) 6, 26 -- Received: from expms6.cites.uiuc.edu (localhost.cites.uiuc.edu [127.0.0.1]) 6, 26 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 6, 26 -- with ESMTP id BGR26023; 6, 26 -- Tue, 28 Mar 2006 15:52:39 -0600 (CST) 6, 26 -- Received: from 128.174.5.212 6, 26 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 6, 26 -- with HTTP/1.1; 6, 26 -- Tue, 28 Mar 2006 14:52:38 -0700 6, 26 -- Date: Tue, 28 Mar 2006 14:52:38 -0700 6, 26 -- From: Changhui LEI {clei-at-uiuc.edu} 6, 26 -- Subject: Re: [Microscopy] Re: Adehsive for Titanium 6, 26 -- To: lkrupp-at-us.ibm.com 6, 26 -- Cc: microscopy-at-microscopy.com 6, 26 -- Reply-To: clei-at-uiuc.edu 6, 26 -- X-Mailer: Webmail Mirapoint Direct 3.4.8-GR 6, 26 -- MIME-Version: 1.0 6, 26 -- Message-Id: {1c12a1f6.a048ceea.8238100-at-expms6.cites.uiuc.edu} 6, 26 -- Content-Type: text/plain; charset=us-ascii 6, 26 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I assume that the time is in hours and that the notation hours.minutes. If so, over a three hour period, your current has varied within a couple percent of its initial value. The "worst" changes came in the first forty minutes, from 14.15 to 14.55, where the intensity fell by 2.1%. After that, it seems that you did something to optimize the column/filament because the intensity came back up and stabilized to within one percent.
Is that good enough for your analyses? I don't know. It probably depends on the measurements you intend to make. It should be good enough for standards. You can go back and re-perform a "quant calibration" more often as you collect standards. That way you should have a good intensity reference for each standard even without a Faraday cup. (For non-ISIS users, I understand that Oxford chose cobalt as a reference material since it was readily available, had K and L peaks detectable in the spectrum, and was not readily subject to oxidation. A "quant calibration" is performed prior to analysis to provide a reference of beam intensity across sessions.)
Another question to consider is that of peak integrals. You have almost 50,000 counts in a peak for a pure element. The standard deviation in that count is 223 or 0.44%, relative. That is roughly on the same order of your beam stability.
However, consider 5% cobalt in an alloy matrix. Your net integral would be 2500 counts with a standard deviation of 50 counts or 2%, relative. That indicates more variability will come from your finite counts than from beam variation. There are also issues of background count.
More detailed and thorough treatments of the error are in the texts on microbeam analysis. However, those texts cannot answer what level of precision or accuracy you require for your analyses.
There are many other issues to be considered before embarking on quantitative analyses. I claim to have performed decent quantitative analysis for years by EDS. Still, I ran into difficulty a few years ago when trying to prepare my own standards to complement our commercially obtained standards. I coated my own standards with carbon and found my totals were off by a couple of percent. It turned out that I had different thicknesses of carbon on my commercial and homemade standards. It made a measurable difference. There are many other issues as well. Be sure to internally check the results of your work to develop confidence in your results.
Warren Straszheim
-----Original Message----- X-from: p.arico-at-email.it [mailto:p.arico-at-email.it] Sent: Monday, March 27, 2006 6:54 PM To: wesaia-at-iastate.edu
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Title-Subject: [Filtered] Lift-out Glass rods
Question: Dear Listeres,
Does anyone of you know the suppliers of lift-out galss rods (1.0 mm in dimameter and solid type)? It will be appreciated if you could kindly share the information.
Free to a good home: LKB Histo Knife maker # 2078. I assume that this is a Ralph knife maker. Some glass too. Very heavy. Yours for he cost of shipping.
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 29 08:56:01 2006 4, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2TEu1wL016916 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Mar 2006 08:56:01 -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 (8.13.6/8.13.6/2.5.1) with ESMTP id k2TEtxvL2998398 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Mar 2006 09:55:59 -0500 4, 22 -- Message-ID: {442A9FFE.5080603-at-ufl.edu} 4, 22 -- Date: Wed, 29 Mar 2006 09:55:58 -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: Ralph knife maker 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 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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Email: dgmorgan-at-ucdavis.edu Name: David Morgan
Organization: UC Davis
Title-Subject: [Filtered] LKB type 7801B Knife Maker
Question: We have inherited an LKB type 7801B glass knife maker that doesn't seem to be working. As a rather general question, can anyone tell me whether it is possible and/or practical to have such an instrument fixed? Any help or suggestions would be welcome, and thanks in advance.
I received many useful answers and observations about Co calibration in Oxford ISIS 300 EDS system. I wish to thank everyone who answered me on this list and on private email, now I am able to do some considerations on the stability of our system and quality of our analyses Pietro
-- ********************************************* Pietro Arico' Dipartimento di Chimica e fisica della Terra ed Applicazioni alle Georisorse ed ai Rischi Naturali (CFTA) University of Palermo VIa Archirafi, 36 Palermo, 90123 - Italy email: p.arico-at-email.it Tel. +390916161574 (ext. 139) *********************************************
==============================Original Headers============================== 4, 18 -- From p.arico-at-email.it Thu Mar 30 00:33:53 2006 4, 18 -- Received: from www.unipa.it (www.unipa.it [147.163.1.5]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2U6Xqe4014656 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 30 Mar 2006 00:33:52 -0600 4, 18 -- Received: from [147.163.124.22] ([147.163.124.22]) 4, 18 -- by www.unipa.it (8.13.1/8.13.1) with ESMTP id k2U6UFnU019707 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 30 Mar 2006 08:30:15 +0200 4, 18 -- Message-ID: {442B7B38.5090909-at-email.it} 4, 18 -- Date: Thu, 30 Mar 2006 08:31:20 +0200 4, 18 -- From: "Pietro Arico'" {p.arico-at-email.it} 4, 18 -- User-Agent: Mozilla Thunderbird 1.5 (Windows/20051201) 4, 18 -- MIME-Version: 1.0 4, 18 -- To: MSA {Microscopy-at-msa.microscopy.com} 4, 18 -- Subject: EDS - Co calibration - thank you! 4, 18 -- Content-Type: text/plain; charset=ISO-8859-15; format=flowed 4, 18 -- Content-Transfer-Encoding: 7bit 4, 18 -- X-Virus-Scanned: ClamAV version 0.88, clamav-milter version 0.87 on www.unipa.it 4, 18 -- X-Virus-Status: Clean ==============================End of - Headers==============================
Hi, we are still using old LKB 7801A Knife Maker and it is still working well. It is a solid device.
Oldrich
Institute of Microbiology Acad. Sci. CR EM Lab Czech Republic
} Email: dgmorgan-at-ucdavis.edu } Name: David Morgan } } Organization: UC Davis } } Title-Subject: [Filtered] LKB type 7801B Knife Maker } } Question: We have inherited an LKB type 7801B glass knife maker that } doesn't seem to be working. As a rather general question, can anyone tell } me whether it is possible and/or practical to have such an instrument } fixed? Any help or suggestions would be welcome, and thanks in advance. }
==============================Original Headers============================== 7, 25 -- From benada-at-biomed.cas.cz Thu Mar 30 01:26:15 2006 7, 25 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2U7QE7Z024694 7, 25 -- for {microscopy-at-microscopy.com} ; Thu, 30 Mar 2006 01:26:14 -0600 7, 25 -- Received: from mail2.biomed.cas.cz (localhost [127.0.0.1]) 7, 25 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k2U7PxJx023232; 7, 25 -- Thu, 30 Mar 2006 09:25:59 +0200 (CEST) 7, 25 -- Received: from 147.231.44.104 7, 25 -- (SquirrelMail authenticated user benada) 7, 25 -- by mail2.biomed.cas.cz with HTTP; 7, 25 -- Thu, 30 Mar 2006 09:25:59 +0200 (CEST) 7, 25 -- Message-ID: {1260.147.231.44.104.1143703559.squirrel-at-mail2.biomed.cas.cz} 7, 25 -- In-Reply-To: {200603300009.k2U09Ynb023609-at-ns.microscopy.com} 7, 25 -- References: {200603300009.k2U09Ynb023609-at-ns.microscopy.com} 7, 25 -- Date: Thu, 30 Mar 2006 09:25:59 +0200 (CEST) 7, 25 -- Subject: Re: [Microscopy] viaWWW: LKB type 7801B Knife Maker 7, 25 -- From: =?iso-8859-2?Q?Old=F8ich_Benada?= {benada-at-biomed.cas.cz} 7, 25 -- To: dgmorgan-at-ucdavis.edu 7, 25 -- Cc: microscopy-at-microscopy.com 7, 25 -- User-Agent: SquirrelMail/1.4.6 7, 25 -- MIME-Version: 1.0 7, 25 -- Content-Type: text/plain;charset=iso-8859-2 7, 25 -- Content-Transfer-Encoding: 8bit 7, 25 -- X-Priority: 3 (Normal) 7, 25 -- Importance: Normal ==============================End of - Headers==============================
We have an (ancient) Hummer 1 coater in our lab that is in good working order, but the argon valve is showing signs of age. It has been replaced in the past, but we do not know of a good source for needle valves of this type. Does anyone have a good source for a replacement, or a salvaged needle valve in a drawer somewhere?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 3, 23 -- From hagglundk1-at-nku.edu Thu Mar 30 12:56:10 2006 3, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2UIuAol019833 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 30 Mar 2006 12:56:10 -0600 3, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 3, 23 -- Thu, 30 Mar 2006 13:56:10 -0500 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 3, 23 -- Content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="us-ascii" 3, 23 -- Subject: Replacement Argon Valve (Hummer 1) 3, 23 -- Date: Thu, 30 Mar 2006 13:56:09 -0500 3, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3586CB-at-mailfac1.hh.nku.edu} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: Replacement Argon Valve (Hummer 1) 3, 23 -- Thread-Index: AcZUK5tAXzEUCjwHSM+7k5PAG7XC1A== 3, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 3, 23 -- To: {microscopy-at-microscopy.com} 3, 23 -- X-OriginalArrivalTime: 30 Mar 2006 18:56:10.0165 (UTC) FILETIME=[9BD8EE50:01C6542B] 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2UIuAol019833 ==============================End of - Headers==============================
You might try swagelok: http://www.swagelok.com/search/find_products.aspx
Search for: metering valve
Regards, Woody White BWXT Services
-----Original Message----- X-from: hagglundk1-at-nku.edu [mailto:hagglundk1-at-nku.edu] Sent: Thursday, March 30, 2006 1:57 PM To: White, Woody N.
We have an (ancient) Hummer 1 coater in our lab that is in good working order, but the argon valve is showing signs of age. It has been replaced in the past, but we do not know of a good source for needle valves of this type. Does anyone have a good source for a replacement, or a salvaged needle valve in a drawer somewhere?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 3, 23 -- From hagglundk1-at-nku.edu Thu Mar 30 12:56:10 2006 3, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2UIuAol019833 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 30 Mar 2006 12:56:10 -0600 3, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 3, 23 -- Thu, 30 Mar 2006 13:56:10 -0500 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 3, 23 -- Content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="us-ascii" 3, 23 -- Subject: Replacement Argon Valve (Hummer 1) 3, 23 -- Date: Thu, 30 Mar 2006 13:56:09 -0500 3, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3586CB-at-mailfac1.hh.nku.edu} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: Replacement Argon Valve (Hummer 1) 3, 23 -- Thread-Index: AcZUK5tAXzEUCjwHSM+7k5PAG7XC1A== 3, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 3, 23 -- To: {microscopy-at-microscopy.com} 3, 23 -- X-OriginalArrivalTime: 30 Mar 2006 18:56:10.0165 (UTC) FILETIME=[9BD8EE50:01C6542B] 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2UIuAol019833 ==============================End of - Headers==============================
==============================Original Headers============================== 15, 27 -- From nwwhite-at-bwxt.com Thu Mar 30 13:23:22 2006 15, 27 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 15, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2UJNMvm029873 15, 27 -- for {microscopy-at-msa.microscopy.com} ; Thu, 30 Mar 2006 13:23:22 -0600 15, 27 -- Received: from ([131.184.13.224]) 15, 27 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.593521; 15, 27 -- Thu, 30 Mar 2006 14:23:11 -0500 15, 27 -- Received: from bwxslynpo01.BWXS.BWXTECH.NET ([131.184.13.4]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 15, 27 -- Thu, 30 Mar 2006 14:23:11 -0500 15, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 27 -- Content-class: urn:content-classes:message 15, 27 -- MIME-Version: 1.0 15, 27 -- Content-Type: text/plain; 15, 27 -- charset="US-ASCII" 15, 27 -- Subject: RE: [Microscopy] Replacement Argon Valve (Hummer 1) 15, 27 -- Date: Thu, 30 Mar 2006 14:23:11 -0500 15, 27 -- Message-ID: {70E4EA352EC987489773D55B0FF83E1C1DF9CB-at-bwxslynpo01.BWXS.BWXTECH.NET} 15, 27 -- X-MS-Has-Attach: 15, 27 -- X-MS-TNEF-Correlator: 15, 27 -- Thread-Topic: [Microscopy] Replacement Argon Valve (Hummer 1) 15, 27 -- Thread-Index: AcZUK8l3Kub/AeK/Q+Sz3HLhe16bdAAA1itA 15, 27 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 15, 27 -- To: {hagglundk1-at-nku.edu} , 15, 27 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 15, 27 -- X-OriginalArrivalTime: 30 Mar 2006 19:23:11.0738 (UTC) FILETIME=[626155A0:01C6542F] 15, 27 -- Content-Transfer-Encoding: 8bit 15, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2UJNMvm029873 ==============================End of - Headers==============================
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Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Filtered] Postdoctoral Position
Question: POST-DOCTORAL POSITION in Transmission Electron Microscopy of Amorphous Thin Films for IR Sensing at The Pennsylvania State University
A postdoctoral position is available in the area of transmission electron microscopy beginning May 1, 2006. The research project focuses on understanding structure and chemistry of amorphous metal-oxides and semiconducting materials for IR sensor applications. Through a variety of electron imaging, spectroscopy and diffraction (e.g. fluctuation EM) techniques we aim to quantify structure and chemistry of amorphous thin films to establish processing/structure/property relationships for this class of materials. Furthermore, we aim to understand the microstructural and microchemical evolution under thermal and electrical stress. Most of the research will be conducted on a JEOL 2010F Field Emission TEM/STEM outfitted with an EDAX energy dispersive x-ray spectrometer (EDS), Gatan Enfina electron energy loss spectrometer (EELS), high-angle annular dark field STEM detector, and acquisition hardware and software for spectrum imaging. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
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Email: dm24-at-cornell.edu Name: David A. Muller
Organization: Cornell university
Title-Subject: [Filtered] Cornell Workshop & Summer School July 13-20 , 2006
Question: On the thirtieth anniversary of the first Cornell workshop on analytical electron microscopy, as well as John Silcox's 45th anniversary at Cornell, the Kavli Institute at Cornell will host a third summer school and workshop from July 13, to July 20, 2006. The conference website is http://www.research.cornell.edu/kic/events/em2006 and a poster is attached.
Electron microscopy has been at the forefront of nanoscale studies of materials, directly probing both physical and electronic properties. From the imaging and spectroscopy of individual dopant atoms and clusters buried inside a semiconductor host to the three-dimensional tomography of nanoparticles and biological structures, and the in-situ observations of nanomechanical deformations and electrodeposition, advances in instrumentation and algorithms have dramatically changed the field of electron microscopy. Early results in sub-angstrom resolution and millivolt spectroscopy are now being applied to materials problems, and international initiatives in aberration-corrected instruments should make such facilities available to the wider community.
The Summer School (July 13-15) will explore general theory of imaging, multislice and Bloch-wave simulations, and the theory of electron energy loss spectroscopy; computer labs are included. The workshop (July 16-20) will assess the present state of analytical electron microscopy and its impact on the physical and biological sciences, and identify the fundamental limits and the new science that next-generation technology should make possible.
Workshop invited speakers include:
Les Allen, University of Melbourne Phil Batson, IBM T.J. Watson Research Center Gianluigi Botton, McMaster University Nigel Browning, LLNL/UC Davis Christian Colliex, CNRS, UniversitÈ Paris Sud John Cummings, University of Maryland Joachim Frank, HHMI, Wadsworth Center Archie Howie, University of Cambridge Ondrej Krivanek, NION Co. Richard Leapman, National Institutes of Health David Muller, Cornell University Harald Rose, Technical University of Darmstadt Frances Ross, IBM, T.J. Watson Research Center John Spence, Arizona State University Suzanne Stemmer, UC Santa Barbara Akira Tonomura, Hitachi Advanced Research Laboratory Maria Varela, Oak Ridge National Laboratory David Williams, Lehigh University Nestor Zaluzec, Argonne National Laboratory
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Email: eoptics-at-mcmaster.ca Name: Fred Pearson
Organization: McMaster University
Title-Subject: [Filtered] TEM-- Double Extraction Replicas for SuperAlloys
Question: I have a user who requires a procedure for looking at super alloy gamma(prime)phase using the double extraction replica method. An procedure from text or from an article would be appreciated. The method involved plastic replica and wax, dissolved in toluene, then carbon coating and shadow casting with Cr.
We are viewing bacteria plated out on filter membranes in the SEM and are having the standard problem of the biofilm/extracellular polysaccharide/mucus/slime around the organisms curdling up during specimen preparation. Normally it's not a huge problem, but this batch of bugs seems to be exceptionally slimy and the stuff is obscuring the structures we want to see.
My question is: are there any techniques for eliminating this substance before or during processing in order to see unBlemished Bugs. As usual, I'm searching for this information in all the usual places, but want to check to see if you all have any pet techniques.
Thanks! Enjoy the weekend.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 11, 23 -- From TindallR-at-missouri.edu Fri Mar 31 13:05:35 2006 11, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 11, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2VJ5ZDo022885 11, 23 -- for {microscopy-at-microscopy.com} ; Fri, 31 Mar 2006 13:05:35 -0600 11, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 11, 23 -- Fri, 31 Mar 2006 13:05:34 -0600 11, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 11, 23 -- Content-class: urn:content-classes:message 11, 23 -- MIME-Version: 1.0 11, 23 -- Content-Type: text/plain; 11, 23 -- charset="us-ascii" 11, 23 -- Subject: SEM: Bacterial Biofilm Blues 11, 23 -- Date: Fri, 31 Mar 2006 13:05:33 -0600 11, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849E69-at-UM-EMAIL09.um.umsystem.edu} 11, 23 -- X-MS-Has-Attach: 11, 23 -- X-MS-TNEF-Correlator: 11, 23 -- Thread-Topic: SEM: Bacterial Biofilm Blues 11, 23 -- thread-index: AcZU9hZwLwb+4wqzTKq02m0f0Q5MmQ== 11, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 11, 23 -- To: {microscopy-at-microscopy.com} 11, 23 -- X-OriginalArrivalTime: 31 Mar 2006 19:05:34.0295 (UTC) FILETIME=[16821670:01C654F6] 11, 23 -- Content-Transfer-Encoding: 8bit 11, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2VJ5ZDo022885 ==============================End of - Headers==============================