We plan to buy a cryo-ultramicrotome for polymer TEM sample preparation. Is anybody familiar with RMC MT-X cryo-ultramicrotome? Any suggestion or any comments for other cryo-ultramicrotome would be greatly appreciated.
Thanks,
Zhiqiang Chen Institute of Advance Materials and New Energy University of Louiville Louisville, KY 40292
==============================Original Headers============================== 4, 26 -- From z0chen09-at-louisville.edu Sun Oct 1 08:33:07 2006 4, 26 -- Received: from erouter2.it-datacntr.louisville.edu (erouter2.it-datacntr.louisville.edu [136.165.237.35]) 4, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k91DX794006592 4, 26 -- for {Microscopy-at-microscopy.com} ; Sun, 1 Oct 2006 08:33:07 -0500 4, 26 -- Received: from PostX2 (postx2.it-servers.louisville.edu [136.165.233.129]) 4, 26 -- by erouter2.it-datacntr.louisville.edu (Postfix) with ESMTP id 8EE957244D 4, 26 -- for {Microscopy-at-microscopy.com} ; Sun, 1 Oct 2006 09:30:25 -0400 (EDT) 4, 26 -- Received: from savgw-out.louisville.edu ([136.165.234.42]) 4, 26 -- by PostX2 (PostX Enterprise 6.0.1 SMTP Adapter) with SMTP ID 421 4, 26 -- for {Microscopy-at-microscopy.com} ; 4, 26 -- Sun, 1 Oct 2006 09:18:18 -0400 (EDT) 4, 26 -- Received: from gwise.louisville.edu ([136.165.232.131]) 4, 26 -- by savgw-out.louisville.edu (SAVSMTP 3.1.7.47) with SMTP id M2006100109294129692 4, 26 -- for {Microscopy-at-microscopy.com} ; Sun, 01 Oct 2006 09:29:41 -0400 4, 26 -- Received: from sgate-MTA by gwise.louisville.edu 4, 26 -- with Novell_GroupWise; Sun, 01 Oct 2006 09:30:25 -0400 4, 26 -- Message-Id: {451F8AA90200005C0000270E-at-gwise.louisville.edu} 4, 26 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 4, 26 -- Date: Sun, 01 Oct 2006 09:30:17 -0400 4, 26 -- From: "Zhiqiang Chen" {z0chen09-at-louisville.edu} 4, 26 -- To: {Microscopy-at-microscopy.com} 4, 26 -- Subject: cryo-ultramicrotome 4, 26 -- Mime-Version: 1.0 4, 26 -- Content-Type: text/plain; charset=US-ASCII 4, 26 -- Content-Transfer-Encoding: 7bit 4, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
X-from: Ken Converse [mailto:kenconverse-at-qualityimages.biz] Sent: Monday, October 02, 2006 11:47 AM To: 'plarson-at-ou.edu'
Hi everyone,
I was wondering if anyone on the board had any advice for venting electron microscope columns/sample chambers. In particular, we want a system that allows the column or sample chamber to reach atmospheric pressure by venting with dry N2 but does not allow the chamber to over pressurize above atmosphere to protect seals, thin window on EDS detectors, etc.
I'm aware that there are on-demand gas regulators that can control this sort of thing. Does anyone have any suggestions for a good vendor or model number for this sort of regulator?
Alternatively, I've also heard that demand valves that divers use are good for this sort of application as it only supplies gas when the diver
breathes or sucks through the mouthpiece (or in this case when the chamber is still under vacuum). Does anyone know a particular brand or type of diver demand valve that works well for this application and is easy to modify (i.e. relatively easy to install standard fittings on the
ends like NPT, etc.)?
Any comments or suggestions would be greatly appreciated. Thanks in advance,
preston
-- Preston Larson Research Scientist Samuel Roberts Noble Electron Microscopy Laboratory 770 Van Vleet Oval Phone: (405) 325-4391
==============================Original Headers============================== 9, 17 -- From plarson-at-ou.edu Wed Sep 27 10:29:15 2006 9, 17 -- Received: from c3p0.ou.edu (mail.zero.ou.edu [129.15.0.75]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8RFTFwv029604 9, 17 -- for {Microscopy-at-Microscopy.Com} ; Wed, 27 Sep 2006 10:29:15 -0500 9, 17 -- Received: from [127.0.0.1] ([129.15.39.56]) 9, 17 -- by c3p0.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 9, 17 -- 2005)) with ESMTP id {0J690035JCCPEI80-at-c3p0.ou.edu} for 9, 17 -- Microscopy-at-Microscopy.Com; Wed, 27 Sep 2006 10:29:13 -0500 (CDT) 9, 17 -- Date: Wed, 27 Sep 2006 10:29:11 -0500 9, 17 -- From: Preston Larson {plarson-at-ou.edu} 9, 17 --
DJ, In November 2000 I donated an ETEC Autoscan to the Red Lion Area Sr. High School in Red Lion, PA along with a very old (vacuum tube logic) Varian vacuum evaporator. They have been using it ever since and I have been providing free service.
This was the school system that my kids went to and the deal was that I would treat them as a full service contract for at least as long as my kids were in the school system. They're all graduated and I've since moved to Maine, but continue to service the scope, just not promising as prompt service due to the current distance involved.
IMHO the problem isn't getting the equipment. It's really not all that unusual for this stuff to come up free. The problem is service. I would encourage anyone who is familiar with servicing SEMs, TEMs and other "high tech" equipment to adopt a school, find them some free equipment and give them free service. If you want to get kids interested in the sciences then they need to have the opportunity to play with the toys that scientists play with. After all, many of us were drawn to our areas of interest by the cool toys.
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: dljones-at-bestweb.net [mailto:dljones-at-bestweb.net] Sent: Thursday, September 28, 2006 10:18 AM To: kenconverse-at-qualityimages.biz
I've been asked several times the following question:
How many high schools in the world actually have equipment such as SEM's
or other fairly sophisticated scientific equipment for use in the classroom?
If you are a high school, or know of a high school that has this kind of
capability, can you please post that information. I would like to know where you are, contact information, and what equipment you use.
I think it will be of interest for our whole scientific community to know the answer to that question, so I suggest that you post the answer to the group as a whole.
If you prefer to answer me directly, please indicate if you prefer information to be shared or not as the case may be.
Thank you,
dj
==============================Original Headers============================== 10, 16 -- From dljones-at-bestweb.net Thu Sep 28 09:15:24 2006 10, 16 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 10, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8SEFONA006752 10, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 09:15:24 -0500 10, 16 -- Received: from localhost ([71.247.114.109]) 10, 16 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 10, 16 -- 9 2005)) with ESMTPA id {0J6B00G7F3LEFSQ0-at-vms044.mailsrvcs.net} for 10, 16 -- Microscopy-at-microscopy.com; Thu, 28 Sep 2006 09:15:18 -0500 (CDT) 10, 16 -- Date: Thu, 28 Sep 2006 10:27:44 -0400 (Eastern Standard Time) 10, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} 10, 16 -- Subject: Perspectives: High School Microscopy 10, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 10, 16 -- To: Microscopy-at-microscopy.com 10, 16 -- Message-id: {Pine.WNT.4.64.0609281008410.3292-at-H-F1} 10, 16 -- MIME-version: 1.0 10, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 26, 23 -- From kenconverse-at-qualityimages.biz Mon Oct 2 11:42:16 2006 26, 23 -- Received: from mta9.adelphia.net (mta9.adelphia.net [68.168.78.199]) 26, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k92GgG6l023112 26, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 2 Oct 2006 11:42:16 -0500 26, 23 -- Received: from Ken ([24.53.5.33]) by mta9.adelphia.net 26, 23 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with ESMTP 26, 23 -- id {20061002164216.XZTT8494.mta9.adelphia.net-at-Ken} ; 26, 23 -- Mon, 2 Oct 2006 12:42:16 -0400 26, 23 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 26, 23 -- To: {dljones-at-bestweb.net} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 26, 23 -- Subject: RE: [Microscopy] Perspectives: High School Microscopy 26, 23 -- Date: Mon, 2 Oct 2006 12:42:03 -0400 26, 23 -- Message-ID: {003801c6e641$b0b59230$6401a8c0-at-Ken} 26, 23 -- MIME-Version: 1.0 26, 23 -- Content-Type: text/plain; 26, 23 -- charset="us-ascii" 26, 23 -- Content-Transfer-Encoding: 7bit 26, 23 -- X-Priority: 3 (Normal) 26, 23 -- X-MSMail-Priority: Normal 26, 23 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 26, 23 -- In-Reply-To: {200609281417.k8SEHkNk011616-at-ns.microscopy.com} 26, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 26, 23 -- Importance: Normal ==============================End of - Headers==============================
a: find an exhaustive listing of the Smithsonian reference materials (and their reference compositions)
b: find out exactly how I can lay my hands, no, my tweezers, on some of them?
I have searched the Smithsonian website but to no avail.
all the best
Ritchie
-- 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, 26 -- From r.sims-at-auckland.ac.nz Mon Oct 2 16:30:31 2006 10, 26 -- Received: from groucho.itss.auckland.ac.nz (mailhost.auckland.ac.nz [130.216.190.11]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k92LUUJN006602 10, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 2 Oct 2006 16:30:31 -0500 10, 26 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) 10, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 6076F352AB 10, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 3 Oct 2006 10:30:29 +1300 (NZDT) 10, 26 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) 10, 26 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 26 -- with ESMTP id 13659-22 for {Microscopy-at-microscopy.com} ; 10, 26 -- Tue, 3 Oct 2006 10:30:29 +1300 (NZDT) 10, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 325CC350CA 10, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 3 Oct 2006 10:30:28 +1300 (NZDT) 10, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 26 -- To: Microscopy-at-microscopy.com 10, 26 -- Date: Tue, 03 Oct 2006 10:35:08 +1300 10, 26 -- MIME-Version: 1.0 10, 26 -- Subject: Smithsonian Microbeam Reference Materials 10, 26 -- Message-ID: {45223CDC.30160.A3CF8C-at-localhost} 10, 26 -- Priority: normal 10, 26 -- X-mailer: Pegasus Mail for Windows (4.21c) 10, 26 -- Content-type: text/plain; charset=US-ASCII 10, 26 -- Content-transfer-encoding: 7BIT 10, 26 -- Content-description: Mail message body 10, 26 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
Gene Jarosowich wrote a short article on the history of the Smithsonian standards in the NIST Journal of Research (proceedings of a NIST-MAS topical workshop that is well worth looking at, about accuracy in epma) in 2002 -- that has an appendix with a list of the standards. You can download that article and others from my probe class web site www.geology.wisc.edu/~johnf/g777/777NISTarticles.html the fifth article down.
John
-- ======================================================== 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, 25 -- From johnf-at-geology.wisc.edu Mon Oct 2 16:57:51 2006 7, 25 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k92Lvobj017480 7, 25 -- for {microscopy-at-microscopy.com} ; Mon, 2 Oct 2006 16:57:50 -0500 7, 25 -- Received: from localhost (localhost [127.0.0.1]) 7, 25 -- by localhost (Postfix) with ESMTP id 7A99B20D08; 7, 25 -- Mon, 2 Oct 2006 16:57:50 -0500 (CDT) 7, 25 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 7, 25 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 7, 25 -- with ESMTP id 05770-09; Mon, 2 Oct 2006 16:57:47 -0500 (CDT) 7, 25 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 7, 25 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 7, 25 -- (No client certificate requested) 7, 25 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 149A320D1C; 7, 25 -- Mon, 2 Oct 2006 16:57:47 -0500 (CDT) 7, 25 -- Mime-Version: 1.0 7, 25 -- Message-Id: {p0623090ac1473a74bfd4-at-[144.92.206.57]} 7, 25 -- In-Reply-To: {200610022136.k92LaZWI014959-at-ns.microscopy.com} 7, 25 -- References: {200610022136.k92LaZWI014959-at-ns.microscopy.com} 7, 25 -- Date: Mon, 2 Oct 2006 16:56:42 -0500 7, 25 -- To: r.sims-at-auckland.ac.nz, microscopy-at-microscopy.com 7, 25 -- From: John Fournelle {johnf-at-geology.wisc.edu} 7, 25 -- Subject: Re: [Microscopy] Smithsonian Microbeam Reference Materials 7, 25 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 25 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
To Owners/Users of Sorvall MT2B or RMC MT2C ultramicrotomes:
Does anyone have any current information regarding third-party repair services in the St. Louis/Kansas City/Chicago area? We need someone that will do the service onsite as we have three or four machines requiring maintenance.
Thank you so much,
Jaclynn Lett Senior Research Technician Research Center for Auditory and Vestibular Studies Department of Otolaryngology Washington University School of Medicine 660 S. Euclid Ave., Campus Box 8115 St. Louis, MO 63110
Voice: 314-747-7257 Fax: 314-747-7230 Email: lettj-at-ent.wustl.edu http://www.otocore.org/ {br/} The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail.
==============================Original Headers============================== 7, 23 -- From LettJ-at-wusm-pcf.wustl.edu Tue Oct 3 11:43:16 2006 7, 23 -- Received: from MAIL4.wusm-pcf.wustl.edu (ip-17-171.wustl.edu [128.252.17.171]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k93GhGPY020654 7, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 3 Oct 2006 11:43:16 -0500 7, 23 -- Received: from EX02.wusm-pcf.wustl.edu ([10.39.162.137]) by MAIL4.wusm-pcf.wustl.edu with Microsoft SMTPSVC(6.0.3790.1830); 7, 23 -- Tue, 3 Oct 2006 11:43:16 -0500 7, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 23 -- Content-class: urn:content-classes:message 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-Type: text/plain; 7, 23 -- charset="us-ascii" 7, 23 -- Subject: EQUIP: ultramicrotome repair 7, 23 -- Date: Tue, 3 Oct 2006 11:43:15 -0500 7, 23 -- Message-ID: {8AADB33205E9CE49BB22275C309FABDA0B5428-at-EX02.wusm-pcf.wustl.edu} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: EQUIP: ultramicrotome repair 7, 23 -- Thread-Index: AcbnCwWoAXNN06/PQuWGMhqvIi2ygQ== 7, 23 -- From: "Lett, Jaclynn" {LettJ-at-ent.wustl.edu} 7, 23 -- To: {Microscopy-at-MSA.Microscopy.Com} 7, 23 -- X-OriginalArrivalTime: 03 Oct 2006 16:43:16.0273 (UTC) FILETIME=[0648D210:01C6E70B] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k93GhGPY020654 ==============================End of - Headers==============================
Colin, I suspect you can get strong thin-section staining of cellulose and other polysaccharides with our favorite, KMnO4 followed by Pb (citrate ok, but we prefer Sato's Lead formula; Essential to avoid NMA (MNA?) in your epoxy mixture; best to use Araldite; my favorite is Araldite 506). I know this Mn-Pb sequence strongly stains the spiral chitin rib in micron and submicron tracheoles of insect muscle, and also stains the dextran (MW 500,000; lovely little wormy threads excluded to the extra-fibrillar space) we've used at 3% to osmotically squeeze the myofilament lattice in the glycerol-detergent-permeabilized muscle fibers. During aqueous fixation, the dextran is lost, rinsed away, but is nicely preserved in situ by freeze-substitution. I think it may even stain, though with less contrast, using the lead stain alone, come to think of it. I seem to recall that lead citrate alone stains glycogen, something I never see any more when working with permeabilized muscle.
Part of our staining success (and certainly our ultimate high contrast) might also be due to our preferred 2-step fixation (works for both aqueous and in -80 acetone for freeze-substitution) using tannic acid followed by uranyl acetate. No additional fix is needed for extracellular structures or well-anchored internal components of permeabilized cells; but you can use OsO4 after TA instead of UrAc. However, OsO4 toxicity and vapor makes it less popular with us than the equally effective UrAc.
I tried Googling got 60 seconds on permanganate staining of cellulose and it looks promising; hit #7 includes "becomes visible as permanganate is a stain for cellulose fibrils ... ". this 1979 reference attributes a 1975 paper DESHPANDE, B. P. (1976). Observations on the fine structure of plant cell walls. I. Use of permanganate staining. Ann. Bot. 40, 433-437.
I leave you to sift through the rest.
-mike reedy-
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- -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
I'm sorry I didn't reply earlier, but if your model of LKB 7801A works like mine then I'm sure this will solve it.
A1. I assume that you have the black handled locking lever to 45 deg forward from vertical and the score selector to || //.
If this doesn't work then it's probably because the clamping head needs loosening off: I hope you have the same diagrams as me so (if not - get back to me): B1. Undo the silver cover (domed flat blade screw head) screw - labelled 7 in Fig 1. B2. using the correct hexagonal allen wrench loosen the screw inside the hole. B3. Now you can loosen the barrel of the locking head by putting something in one of 4 holes around the cylinder at the base of the shaft for the black handled locking lever. Turn it one or two rotations clockwise and you should see it loosen a bit.
Now if you go back to A1 above it should be possible to remove the head easily because it is not clamped so tightly.
If this works remember to re-adjust the barrel tension (B3 above) so that the black handled locking lever is clamped at about 90 to 100 deg from vertical. Effectively do B3, B2 and B1 in reverse.
Please come back to me if my instructions are too vague or vary from your model.
I have copied this to the microscopy list just in case anyone else has this problem.
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: benada-at-biomed.cas.cz
Hi, All: Please let me know if any body has a second TEM with CCD for sale. Thank you for your attention.
Zhiqiang Chen Institute for Advance Materials and Renewable Energy University of Louisville Louisville, KY 40292
==============================Original Headers============================== 2, 26 -- From z0chen09-at-louisville.edu Wed Oct 4 16:01:44 2006 2, 26 -- Received: from erouter2.it-datacntr.louisville.edu (erouter2.it-datacntr.louisville.edu [136.165.237.35]) 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k94L1iVY017989 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 4 Oct 2006 16:01:44 -0500 2, 26 -- Received: from PostX2 (postx2.it-servers.louisville.edu [136.165.233.129]) 2, 26 -- by erouter2.it-datacntr.louisville.edu (Postfix) with ESMTP id 3E8BA71057 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 4 Oct 2006 16:57:22 -0400 (EDT) 2, 26 -- Received: from savgw-out.louisville.edu ([136.165.234.42]) 2, 26 -- by PostX2 (PostX Enterprise 6.0.1 SMTP Adapter) with SMTP ID 840 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; 2, 26 -- Wed, 4 Oct 2006 16:45:06 -0400 (EDT) 2, 26 -- Received: from gwise.louisville.edu ([136.165.232.131]) 2, 26 -- by savgw-out.louisville.edu (SAVSMTP 3.1.7.47) with SMTP id M2006100416564801610 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 04 Oct 2006 16:56:48 -0400 2, 26 -- Received: from sgate-MTA by gwise.louisville.edu 2, 26 -- with Novell_GroupWise; Wed, 04 Oct 2006 16:57:22 -0400 2, 26 -- Message-Id: {4523E7F5.B28C.005C.0-at-gwise.louisville.edu} 2, 26 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 2, 26 -- Date: Wed, 04 Oct 2006 16:57:08 -0400 2, 26 -- From: "Zhiqiang Chen" {z0chen09-at-louisville.edu} 2, 26 -- To: {Microscopy-at-MSA.Microscopy.Com} 2, 26 -- Subject: looking for second-hand TEM 2, 26 -- Mime-Version: 1.0 2, 26 -- Content-Type: text/plain; charset=US-ASCII 2, 26 -- Content-Transfer-Encoding: 7bit 2, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
I would like to look at actin structure in fish skin epithelial cells. I usually use 2%GA-2%PFA fixative and spurr's resin, but I think actin structure is not clear enough. If anyone knows or experienced well preserved morphology, please give me any advice.
Thank you so much.
--------------------- Fumi KATOH post doctoral research fellow St. Francis Xavier University fkatoh-at-stfx.ca
==============================Original Headers============================== 5, 20 -- From fkatoh-at-stfx.ca Wed Oct 4 18:34:47 2006 5, 20 -- Received: from xmail1.stfx.ca (xmail1.stfx.ca [141.109.221.24]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k94NYjUB031545 5, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 4 Oct 2006 18:34:46 -0500 5, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 20 -- Content-class: urn:content-classes:message 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="iso-8859-1" 5, 20 -- Subject: TEM-fixation for actin 5, 20 -- Date: Wed, 4 Oct 2006 20:32:43 -0300 5, 20 -- Message-ID: {D1DA484CA5D7D5499AB6BBB4BCA429175CA4EC-at-demetrius.ad.stfx.ca} 5, 20 -- X-MS-Has-Attach: 5, 20 -- X-MS-TNEF-Correlator: 5, 20 -- Thread-Topic: TEM-fixation for actin 5, 20 -- Thread-Index: AcboDWPzpDtFkkVpQOiZMlr4+eES/g== 5, 20 -- From: "Fumi Katoh" {fkatoh-at-stfx.ca} 5, 20 -- To: {Microscopy-at-Microscopy.Com} 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k94NYjUB031545 ==============================End of - Headers==============================
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 marie.johnson1-at-att.net as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: marie.johnson1-at-att.net Name: Marie
Organization: Continental Carbon Company
Title-Subject: [Filtered] CamScan Model Cs44
Question: Does anyone have info on this SEM? Specs/year of manufacture? Thank you, Marie
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Email: burrmich-at-msu.edu Name: Mike
Title-Subject: [Filtered] SEM (filament/image issues)
Question: Please help a newbie out a little...
Our regular SEM operator isn't here anymore (can't contact them) so i'm now in charge of running it. (used it sparingly in college)
We have a Hitachi S-570 (yeah a very old one). When i first started it up after about a month downtime, the filament was burnt. So i replaced the filament according to the settings in the manual. Well after starting it up again, i used the filament image mode to find the saturation point and everything. my settings were 20KV and WD of 30mm. I have a fairly circular filament image when the current gets up to only 60 micro amps... then there is a totally seperate half circle pertruding out from the side of the screen. When i turn the fillament knob to get a current above about 80, the image stays the same shape but starts to dissapear.
So when i go back to the best filament image (only 50 micro amps) and switch to normal mode to see an image, i can only see a very faint outline of my specimen. Focusing and gun allignments do nothing.
I'm pretty sure my problem has to do with when i set the new filament, but it was done exactly by the manual.
Electron/Ion Beam Instrument Engineer The University of Oregon's Center for Advanced Materials Characterization in Oregon (CAMCOR) is seeking applications for a full time staff position to begin January 2007. A strong background in maintaining, trouble shooting and upgrading electron/ion beam instruments and associated high voltage, vacuum, mechanical and electrical systems is required. Experience x-ray diffraction instrumentation is also desirable.
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The successful candidate with have a BS in a beam microscopy related field and an extensive background in instrument field service with significant practical experience troubleshooting high vacuum electron and ion beam instrumentation at both the system and PC board levels. Must be able to read and understand schematics for electronic circuits and systems. The successful applicant will be involved in modifying/improving instrumentation capabilities to enable the equipment to more fully support unique research needs and will be expected to work intimately with the scientific staff and research faculty. We seek candidates with a demonstrated commitment to working effectively with students, faculty and staff from diverse backgrounds.
Interested persons should send a resume with a detailed description of work experience and skills, and arrange for two letters of recommendation to be sent to: CAMCOR Instrument Engineer Search Committee, 1253 University of Oregon, Eugene, OR, 97403-1253. To be assured of full consideration, application materials must be received by November 1, 2006, but the search will remain open until the position is filled. For further information, contact John Donovan (donovan-at-uoregon.edu).
University of Oregon is an AA/EEO employer committed to cultural diversity.
==============================Original Headers============================== 6, 21 -- From donovan-at-uoregon.edu Thu Oct 5 12:31:47 2006 6, 21 -- Received: from smtp.uoregon.edu (mserv3.uoregon.edu [128.223.142.101]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95HVkqZ016427 6, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 12:31:47 -0500 6, 21 -- Received: from Probev.uoregon.edu (probev.uoregon.edu [128.223.92.75]) 6, 21 -- (authenticated bits=0) 6, 21 -- by smtp.uoregon.edu (8.13.7/8.13.7) with ESMTP id k95HVkVn032595 6, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 6, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 10:31:46 -0700 6, 21 -- Message-Id: {7.0.1.0.0.20061005103033.00f89840-at-uoregon.edu} 6, 21 -- Message-Id: {7.0.1.0.0.20061004112624.039222b0-at-uoregon.edu} 6, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 6, 21 -- Date: Thu, 05 Oct 2006 10:30:39 -0700 6, 21 -- To: microscopy-at-microscopy.com 6, 21 -- From: John Donovan {donovan-at-uoregon.edu} 6, 21 -- Subject: Full Time Instrument Engineer Position at the University of 6, 21 -- Oregon 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 21 -- X-Virus-Scanned: ClamAV 0.88.4/1998/Thu Oct 5 07:55:28 2006 on mserv3 6, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I am happy to announce that DVDs of the 2006 MSA tutorials are now available. John Mansfield and his crew have done a great job once again. For full information about purchasing these DVDs or any others in the collection, please go to the MSA web site (www.microscopy.org) and look under REFERENCE for the video library.
Greg Erdos
==============================Original Headers============================== 3, 21 -- From gwe-at-ufl.edu Thu Oct 5 13:31:00 2006 3, 21 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 3, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95IUx4d027920 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 13:31:00 -0500 3, 21 -- Received: from [10.228.0.31] (ssrb-vpn1-0-31.vpn.ufl.edu [10.228.0.31]) 3, 21 -- (authenticated bits=0) 3, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k95IUvCq3047440 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 14:30:58 -0400 3, 21 -- Message-ID: {4525798A.8090607-at-ufl.edu} 3, 21 -- Date: Thu, 05 Oct 2006 14:30:50 -0700 3, 21 -- From: greg Erdos {gwe-at-ufl.edu} 3, 21 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 3, 21 -- MIME-Version: 1.0 3, 21 -- To: microscopy-at-microscopy.com 3, 21 -- Subject: 2006 Tutorials Available 3, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 21 -- Content-Transfer-Encoding: 7bit 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==============================
I am interested in gathering advice from members of the list serve about lighting for our new FE TEM. We currently have a 3X3 grid of fluorescent lights for general lighting, and servicing the instrument etc. We've installed clean electrical service - back to the building ground with good wiring and checked for fields. In particular, I'd be interested in hearing from members about their experience with spot lighting installations to illuminate the TEM and workstations while in operation. In the past we have just installed track lighting with high intensity halogen spotlights mounted in movable heads to direct the light where we need the illumination. The spotlights were controlled with a dimmer switch to adjust the intensity to the desired level. I understand that mounting the dimmer switch to the microscope can lead to ground loop problems. What I'd like to hear is if anybody has experience with new installations, things to watch out for (ground loops, fields that effect the microscope etc.), and also any recomendations for lighting systems and or components. Thanks in advance,
Greg
-- -- ================================================================== Greg Strout Research Scientist, University of Oklahoma Past President, Oklahoma Microscopy Society (OMS) e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
==============================Original Headers============================== 6, 19 -- From gstrout-at-ou.edu Thu Oct 5 13:53:43 2006 6, 19 -- Received: from fourlom.ou.edu (mail.zero.ou.edu [129.15.0.75]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95IrhfC006506 6, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 13:53:43 -0500 6, 19 -- Received: from [127.0.0.1] ([129.15.38.202]) 6, 19 -- by fourlom.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 6, 19 -- 28 2005)) with ESMTP id {0J6O0050OF5G6W10-at-fourlom.ou.edu} for 6, 19 -- Microscopy-at-microscopy.com; Thu, 05 Oct 2006 13:53:41 -0500 (CDT) 6, 19 -- Date: Thu, 05 Oct 2006 13:53:37 -0500 6, 19 -- From: Greg Strout {gstrout-at-ou.edu} 6, 19 -- Subject: Room lighting and new microscope installation 6, 19 -- To: Microscopy-at-microscopy.com 6, 19 -- Message-id: {452554B1.40509-at-ou.edu} 6, 19 -- MIME-version: 1.0 6, 19 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-transfer-encoding: 7BIT 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 19 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
On Oct 5, 2006, at 11:53 AM, gstrout-at-ou.edu wrote:
} I am interested in gathering advice from members of the list serve } about } lighting for our new FE TEM. } We currently have a 3X3 grid of fluorescent lights for general } lighting, } and servicing the instrument etc. We've installed clean electrical } service - back to the building ground with good wiring and checked for } fields. } In particular, I'd be interested in hearing from members about their } experience with spot lighting installations to illuminate the TEM and } workstations while in operation. In the past we have just installed } track lighting with high intensity halogen spotlights mounted in } movable } heads to direct the light where we need the illumination. The } spotlights were controlled with a dimmer switch to adjust the intensity } to the desired level. } I understand that mounting the dimmer switch to the microscope can lead } to ground loop problems. } What I'd like to hear is if anybody has experience with new } installations, things to watch out for (ground loops, fields that } effect } the microscope etc.), and also any recomendations for lighting systems } and or components. } Dear Greg, We have two fixtures each with two 4' fluorescent lights and six track lighting fixtures with 50 W halogen flood lights. The fluorescents are controlled by either a wall switch or a switch mounted on the scope, and the halogens are controlled by a dimmer mounted on the scope--the wires to the switches are run through conduit along the scope frame. We have not seen any adverse effect from the scope-mounted switches on either our FEI T12 or FEI TF30H. 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 Oct 5 15:05:13 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 (8.12.11.20060308/8.12.8) with ESMTP id k95K5DJn018514 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 15:05:13 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 835D835456 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 13:05:12 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 420D210A0D4 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 13:05:11 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200610051853.k95IrnNq006665-at-ns.microscopy.com} 4, 22 -- References: {200610051853.k95IrnNq006665-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {37d161280f0a3ff39290d89266cf039a-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Room lighting and new microscope installation 4, 22 -- Date: Thu, 5 Oct 2006 13:09:08 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
My lab is setup somewhat like yours, with fluorescents and incandescent track lighting.
I don't have a dimmer, but the track lighting is low brightness. When I need to see something under high light conditions, I use the fluorescents. To make this easy, I have a switch at the operating position (RF/wireless) to control the fluorescents. The track lights are on all the time.
Dimmers should not generate ground loops. However, most are pretty good at generating electrical noise that can be radiated, conveyed over the power line, or both. This may or may not be a problem depending on the dimmer quality, the lab layout and the susceptibility of the TEM. A noise free dimmer is a variable autotransformer (Variac TM). Nothing is perfect... They can generate magnetic fields. Fields tend to be small, however, due to the steel shielding and the torrodial core.
Another caveat... New high efficiency fluorescents use solid state switching devices instead of the old saturatable transformer "ballasts". If poorly designed, these can also generate electrical noise. FWIW, The ones I have (brand unknown) do not affect my SEM.
Regards, Woody BWXT Services
-----Original Message----- X-from: gstrout-at-ou.edu [mailto:gstrout-at-ou.edu] Sent: Thursday, October 05, 2006 2:54 PM To: White, Woody N.
Hi everyone,
I am interested in gathering advice from members of the list serve about
lighting for our new FE TEM. We currently have a 3X3 grid of fluorescent lights for general lighting,
and servicing the instrument etc. We've installed clean electrical service - back to the building ground with good wiring and checked for fields. In particular, I'd be interested in hearing from members about their experience with spot lighting installations to illuminate the TEM and workstations while in operation. In the past we have just installed track lighting with high intensity halogen spotlights mounted in movable
heads to direct the light where we need the illumination. The spotlights were controlled with a dimmer switch to adjust the intensity to the desired level. I understand that mounting the dimmer switch to the microscope can lead to ground loop problems. What I'd like to hear is if anybody has experience with new installations, things to watch out for (ground loops, fields that effect
the microscope etc.), and also any recomendations for lighting systems and or components. Thanks in advance,
Greg
-- -- ================================================================== Greg Strout Research Scientist, University of Oklahoma Past President, Oklahoma Microscopy Society (OMS) e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
==============================Original Headers============================== 6, 19 -- From gstrout-at-ou.edu Thu Oct 5 13:53:43 2006 6, 19 -- Received: from fourlom.ou.edu (mail.zero.ou.edu [129.15.0.75]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95IrhfC006506 6, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 13:53:43 -0500 6, 19 -- Received: from [127.0.0.1] ([129.15.38.202]) 6, 19 -- by fourlom.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 6, 19 -- 28 2005)) with ESMTP id {0J6O0050OF5G6W10-at-fourlom.ou.edu} for 6, 19 -- Microscopy-at-microscopy.com; Thu, 05 Oct 2006 13:53:41 -0500 (CDT) 6, 19 -- Date: Thu, 05 Oct 2006 13:53:37 -0500 6, 19 -- From: Greg Strout {gstrout-at-ou.edu} 6, 19 -- Subject: Room lighting and new microscope installation 6, 19 -- To: Microscopy-at-microscopy.com 6, 19 -- Message-id: {452554B1.40509-at-ou.edu} 6, 19 -- MIME-version: 1.0 6, 19 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-transfer-encoding: 7BIT 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 19 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
==============================Original Headers============================== 24, 26 -- From nwwhite-at-bwxt.com Thu Oct 5 15:15:51 2006 24, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 24, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95KFo7O029126 24, 26 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 15:15:51 -0500 24, 26 -- Received: from ([131.184.13.224]) 24, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2375910; 24, 26 -- Thu, 05 Oct 2006 16:15:32 -0400 24, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 24, 26 -- Thu, 5 Oct 2006 16:15:32 -0400 24, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 24, 26 -- Content-class: urn:content-classes:message 24, 26 -- MIME-Version: 1.0 24, 26 -- Content-Type: text/plain; 24, 26 -- charset="US-ASCII" 24, 26 -- Subject: RE: [Microscopy] Room lighting and new microscope installation 24, 26 -- Date: Thu, 5 Oct 2006 16:15:32 -0400 24, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D098-at-BWXSPO01.BWXS.BWXTECH.NET} 24, 26 -- X-MS-Has-Attach: 24, 26 -- X-MS-TNEF-Correlator: 24, 26 -- Thread-Topic: [Microscopy] Room lighting and new microscope installation 24, 26 -- Thread-Index: Acbor7FTkYpsjxQGQq6Azv3EOIESJgAB+2ow 24, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 24, 26 -- To: {gstrout-at-ou.edu} , "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 24, 26 -- X-OriginalArrivalTime: 05 Oct 2006 20:15:32.0570 (UTC) FILETIME=[028903A0:01C6E8BB] 24, 26 -- Content-Transfer-Encoding: 8bit 24, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k95KFo7O029126 ==============================End of - Headers==============================
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Email: ngohad-at-clemson.edu Name: Neeraj V. Gohad
Organization: Dept. Biological Sciences, Clemson University
Title-Subject: [Filtered] Confocal Microscopy
Question: Dear All,
I am going to purchase some DVDs and Tapes from MSA on various topics in microscopy. Could any one please recommend a DVD or tape from MSAs list on Confocal microscopy. There are a buch on confocal microscopy. I am getting to be an advanced user so I want something that will lecture on advanced techniques.
Please let me know,
Thank you in advance,
Neeraj V. Gohad Graduate Research Assistant, Dept. Biological Sciences, Clemson University.
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Email: dang-at-oakland.edu Name: Loan Dang
Organization: Oakland University
Title-Subject: [Filtered] The protocol for fixing the plant sample
Question: I would like to have the standard procedure for fixing the plant material. I need to fix the maple leaf sample. Thanks LD
Hello, I am so very grateful to all who send me advices and hints how to solve our trouble with the knifemaker. Our LKB 7801A is now working fine. Thanking you, again.
My best regards from Prague Oldrich
==============================Original Headers============================== 2, 20 -- From benada-at-biomed.cas.cz Fri Oct 6 01:43:19 2006 2, 20 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k966hI6o007213 2, 20 -- for {microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 01:43:18 -0500 2, 20 -- Received: from [147.231.44.101] (u117ob.mbu.cas.cz [147.231.44.101]) 2, 20 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k966dBI0009934 2, 20 -- for {microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 08:39:11 +0200 (CEST) 2, 20 -- From: benada-at-biomed.cas.cz 2, 20 -- To: microscopy-at-microscopy.com 2, 20 -- Date: Fri, 06 Oct 2006 08:43:14 +0200 2, 20 -- MIME-Version: 1.0 2, 20 -- Subject: LKB knifemaker 7801A problem - Thanks 2, 20 -- Message-ID: {45261722.18601.161A14-at-benada.biomed.cas.cz} 2, 20 -- Priority: normal 2, 20 -- X-mailer: Pegasus Mail for Windows (4.41) 2, 20 -- Content-type: text/plain; charset=US-ASCII 2, 20 -- Content-transfer-encoding: 7BIT 2, 20 -- Content-description: Mail message body 2, 20 -- X-Antivirus: avast! (VPS 0640-3, 05.10.2006), Outbound message 2, 20 -- X-Antivirus-Status: Clean ==============================End of - Headers==============================
Thanks to all those who replied to my query regarding the sectioning of cotton fibres for TEM. As usual the replies were exceptionally useful and have given us a number of leads to follow.
Cheers and have a good weekend.
Colin Veitch
Electron Microscopist CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia. E-mail: colin.veitch-at-csiro.au Web: http://www.tft.csiro.au
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
==============================Original Headers============================== 8, 28 -- From Colin.Veitch-at-csiro.au Fri Oct 6 01:52:44 2006 8, 28 -- Received: from vic-MTAout2.csiro.au (vic-MTAout2.csiro.au [150.229.64.38]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k966qhA9017786 8, 28 -- for {microscopy-at-msa.microscopy.com} ; Fri, 6 Oct 2006 01:52:44 -0500 8, 28 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=MJfZXFYeG9il5Dear5zfD95j+OvuR3g38phqv+vcGJkMUpCIOxR6YB1ge+vn9V59t7fqCU9/UciOFHugqzo8ip/BSkUOsGZNniCYxEoQIUB47oIZ1BUrI/YiwCUVqquE; 8, 28 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 8, 28 -- by vic-MTAout2.csiro.au with ESMTP; 06 Oct 2006 16:52:43 +1000 8, 28 -- X-IronPort-AV: i="4.09,268,1157292000"; 8, 28 -- d="scan'208"; a="97313713:sNHT25809548" 8, 28 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 28 -- Fri, 6 Oct 2006 16:52:36 +1000 8, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 28 -- content-class: urn:content-classes:message 8, 28 -- MIME-Version: 1.0 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="us-ascii" 8, 28 -- Subject: Cotton sectioning thanks 8, 28 -- Date: Fri, 6 Oct 2006 16:52:35 +1000 8, 28 -- Message-ID: {32CDDDAA7161394599F00254949157490248E3-at-exvic5-gex.nexus.csiro.au} 8, 28 -- X-MS-Has-Attach: 8, 28 -- X-MS-TNEF-Correlator: 8, 28 -- Thread-Topic: Cotton sectioning thanks 8, 28 -- Thread-Index: AcbpE/3rE6/8kEF8QQWwfZ6SLEINnw== 8, 28 -- From: {Colin.Veitch-at-csiro.au} 8, 28 -- To: {microscopy-at-msa.microscopy.com} 8, 28 -- X-OriginalArrivalTime: 06 Oct 2006 06:52:36.0202 (UTC) FILETIME=[019864A0:01C6E914] 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k966qhA9017786 ==============================End of - Headers==============================
Hi Dang, Your question is impossible to answer without more information. What do you want to observe in the plant, and with what type of microscopy? The choice of protocol is guided by the answers to those questions.
By the way, to find a lot of information about this in a small space, search your library catalog for "microtechnique". That word sounds like it means delicate surgery or something but in fact it is what books about fixation protocols are often titled. There probably are one or two at your library (try Plant or Botanical Microtechnique).
Good luck, Tobias
} Organization: Oakland University } } Title-Subject: [Filtered] The protocol for fixing the plant sample } } Question: I would like to have the standard procedure for fixing the } plant material. } I need to fix the maple leaf sample. } Thanks } LD } } -
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (brookie_cookie0-at-hotmai.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, October 5, 2006 at 09:19:36 ---------------------------------------------------------------------------
Looking for methods or experiences in measuring the total surface area of 1 gram of mineral (diopside) powder with grain sizes between 150 and 250 microns. I have heard of SEM techniques but not sure how that is done. Any Ideas?
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 6, 23 -- From rbeavers-at-mail.smu.edu Fri Oct 6 15:06:07 2006 6, 23 -- Received: from s31xua.systems.smu.edu (s31xua.systems.smu.edu [129.119.70.72]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k96K66Gd007881 6, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 15:06:07 -0500 6, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xua.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Fri, 6 Oct 2006 15:06:04 -0500 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: Measurement of surface area 6, 23 -- Date: Fri, 6 Oct 2006 15:06:04 -0500 6, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB235E26-at-s31xe7.systems.smu.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: Measurement of surface area 6, 23 -- Thread-Index: AcbpgtoCt0jPMxFLTnOwa3b2evFx2g== 6, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 6, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 06 Oct 2006 20:06:05.0119 (UTC) FILETIME=[DAB89CF0:01C6E982] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k96K66Gd007881 ==============================End of - Headers==============================
The University of Wisconsin Milwaukee is searching for a full time probationary senior instrumentation specialist to train and assist faculty, research and technical staff and students in use of electron and atomic force microscopes, vacuum systems, and specimen preparation instruments; to install, operate, update, maintain and repair scientific instruments; to administer instrument use and enforce lab and chemical safety protocols. Candidates must have a Bachelor's degree with course work in physical or biological sciences, a post-bachelors certificate or degree in electron microscopy; at least four years of experience in microscopy and /or surface science; ability to effectively operate research instruments and computers; strong written and verbal communication skills to work with lab users from diverse backgrounds. Well qualified applicants will have training and/or experience with HRTEM, AFM, SQUID, vacuum leak checkers, preparation of surface and TEM specimen. Minimum salary of $46,191, commensurate with qualifications. UWM is an AA/EEO employer
To apply, send letter of intent and resume with three references, postmarked by Nov. 8, 2006, to: Search and Screen Chair - Microscopy, UWM L&S Holton 249, P. O. Box 413, Milwaukee, WI 53201-0413, For more information please see www.uwm.edu/letsci/jobs
Marija Gajdardziska-Josifovska Associate Dean for Natural Sciences University of Wisconsin Milwaukee tel: 414 229 2925 fax 414 229 6827
==============================Original Headers============================== 4, 24 -- From marijagj-at-gmail.com Fri Oct 6 16:49:53 2006 4, 24 -- Received: from nf-out-0910.google.com (nf-out-0910.google.com [64.233.182.188]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k96Lnq7j019806 4, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 6 Oct 2006 16:49:53 -0500 4, 24 -- Received: by nf-out-0910.google.com with SMTP id c2so848197nfe 4, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 06 Oct 2006 14:49:51 -0700 (PDT) 4, 24 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 24 -- s=beta; d=gmail.com; 4, 24 -- h=received:message-id:date:from:to:subject:cc:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 24 -- b=rs8qh+vc/Pr73QSxeHHY9YaCfsiYJHuwU0K+8cHSLdjrdl1I6kCv15/14qIPaKlYHzPVnIi9tPTs2wiZGkL3wiu6WInnpSXY5Umpbz9cPBlF57zN55x61zkoLykCpo4UugIymVRRqCX9dFwPvTNMUT2wTaTIVyRIrPzD1lihg4M= 4, 24 -- Received: by 10.49.8.1 with SMTP id l1mr6060288nfi; 4, 24 -- Fri, 06 Oct 2006 14:49:51 -0700 (PDT) 4, 24 -- Received: by 10.49.87.12 with HTTP; Fri, 6 Oct 2006 14:49:51 -0700 (PDT) 4, 24 -- Message-ID: {ee4af5560610061449x27fa2469x25e6a2d6292e058b-at-mail.gmail.com} 4, 24 -- Date: Fri, 6 Oct 2006 16:49:51 -0500 4, 24 -- From: "Marija Gajdardziska-Josifovska" {marijagj-at-gmail.com} 4, 24 -- To: Microscopy-at-Microscopy.Com 4, 24 -- Subject: Senior Instrumentation Specialist Position 4, 24 -- Cc: "Linda Daley" {lindaley-at-uwm.edu} , 4, 24 -- "Marija Gajdardziska-Josifovska" {mgj-at-uwm.edu} 4, 24 -- MIME-Version: 1.0 4, 24 -- Content-Type: text/plain; charset=UTF-8; format=flowed 4, 24 -- Content-Transfer-Encoding: 7bit 4, 24 -- Content-Disposition: inline ==============================End of - Headers==============================
Version 2.0.0 of the 2dx software for image-processing for 2D crystals is now available here: http://2dx.org/download/2dx-software
This version contains numerous new features, and has included all the bugfixes, suggestions, and better algorithms that were brought to us from several users.
A 13-minute movie that demonstrates the basic usage of 2dx is available here: http://2dx.org/documentation/2dx-software/tutorial/2dx_image-primer
A description of 2dx in JSB can be found here: http://dx.doi.org/10.1016/j.jsb.2006.07.020
Bryant, Xiangyan and Henning.
Henning Stahlberg, Molecular & Cellular Biology, Briggs Hall 5, University of California at Davis, 1 Shields Ave., Davis, CA 95616, USA Tel: +1-530-752 8282 (office), +1-530-754 8285 (lab) Fax: +1-530-752 3085 mailto:HStahlberg-at-ucdavis.edu http://stahlberglab.ucdavis.edu
==============================Original Headers============================== 11, 18 -- From HStahlberg-at-ucdavis.edu Fri Oct 6 19:15:05 2006 11, 18 -- Received: from pop23.ucdavis.edu (pop23.ucdavis.edu [169.237.105.13]) 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k970F4kc032124 11, 18 -- for {Microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 19:15:05 -0500 11, 18 -- Received: from [169.237.214.65] ([169.237.214.65]) 11, 18 -- by pop23.ucdavis.edu (8.13.7/8.13.1/it-std-5.2.0) with ESMTP id k970Dsxk000955; 11, 18 -- Fri, 6 Oct 2006 17:15:04 -0700 (PDT) 11, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 11, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 11, 18 -- Message-Id: {D71030DD-B067-4DBE-8B42-B998585F0F94-at-ucdavis.edu} 11, 18 -- Cc: "Bryant R. Gipson" {BRGipson-at-ucdavis.edu} , 11, 18 -- Xiangyan Zeng {XZeng-at-ucdavis.edu} 11, 18 -- Content-Transfer-Encoding: 7bit 11, 18 -- From: Henning Stahlberg {HStahlberg-at-ucdavis.edu} 11, 18 -- Subject: 2dx - Image Processing for 2D Crystal Images: Version 2.0.0 now available 11, 18 -- Date: Fri, 6 Oct 2006 17:15:04 -0700 11, 18 -- To: Microscopy-at-microscopy.com 11, 18 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (JawaharMKD-at-aol.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, October 6, 2006 at 17:33:58 ---------------------------------------------------------------------------
Email: JawaharMKD-at-aol.com Name: Sonia
Organization: Queen Mary
Education: Undergraduate College
Location: London, UK
Question: What is the mechanism for simple staining? How does Gram staining actually work? What is the principle behind streaking on a plate? Why do we use selective or differential media?
} Just wanted to ask for your recommendation of a good } brand/model of UPS for an SEM system. } } We have a Phillips/FEI Quanta 200 ESEM and we are thinking of } buying an uninterrupted power supply for voltage surge protection.
Our Quanta 400 is getting along well with a Powerware 9125 6000VA. I sometimes wish it offered a little more headroom because the initial pumpdown sequence will infrequently trigger an overload warning, but under normal operating conditions, 24/7, it is fine. Powerware support is excellent. They replaced a UPS that was frequently giving me overload warnings with a UPS that subsequently hasn't issued a warning yet. Ours also supports 2 computers along with an EDX system, and if your installation is like mine, there's no need to support a chiller, so the same KVA may be all you need.
It is also the case that our UPS, with only a pair of batteries, is only intended to save power for relatively short 15 minute outages, which covers ~99% of our problems. If you want extended operation, then Powerware offers additional battery modules for this same model.
A UPS this size can be noisy, and possibly emit detrimental EM radiation. Ours is in a different room along with the 208-to-230V step-up transformer. 230V is the input voltage for the UPS.
See: http://www.powerware.com/UPS/9125RM_UPS.asp
HTH, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
==============================Original Headers============================== 10, 20 -- From michael-at-shaffer.net Mon Oct 9 08:48:56 2006 10, 20 -- Received: from n007.sc1.cp.net (smtpout1453.sc1.he.tucows.com [64.97.157.153]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k99DmtTb027557 10, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 9 Oct 2006 08:48:55 -0500 10, 20 -- Received: from rarewolf (205.251.84.119) by n007.sc1.cp.net (7.2.069.1) (authenticated as michael-at-shaffer.net) 10, 20 -- id 452428650012157A; Mon, 9 Oct 2006 13:48:50 +0000 10, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 10, 20 -- To: {mmiralles-at-pi.ac.ae} 10, 20 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 10, 20 -- Subject: RE: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 10, 20 -- Date: Mon, 9 Oct 2006 11:18:52 -0230 10, 20 -- Message-ID: {009201c6eba9$a8b8c6c0$4701a8c0-at-rarewolf} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="US-ASCII" 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- X-Mailer: Microsoft Office Outlook 11 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 10, 20 -- Thread-Index: AcbrhcqqoOqJZ/dISOGHCQh0ezXlKAAIRzQQ 10, 20 -- In-Reply-To: {200610090932.k999Wind013147-at-ns.microscopy.com} ==============================End of - Headers==============================
If your concern is surges or sags in power supply, you don't need an UPS. What you would need is a stabilizer like Topaz makes. This is a magnetic resonance system that adjusts output voltage to be at specified output. It also includes input spike protection and EMI/RFI removal.
If you do want an UPS, and these are very nice to have, there are several makers. I use two Liebert Nfinity 8KVA units with redundant power and control modules. SEM and its PC are on one UPS while EDS, EBSD and other PCs and chiller and compressor are on another one. Each UPS runs at about 25% total capacity and will run the whole system for about 250 minutes if power fails--which it usually does in winter and summer here in California. These units regulate output voltage to within about 2% of specified value. They are double conversion in that they take in AC, convert it to DC and invert the DC to AC output. When input AC fails, DC from the batteries is inverted to AC output. Along with this is spike protection, sag protection and EMI/RFI protection.
These units work on 208-240VAC input and output similar voltage. I take 240VAC in and output 240VAC and 120VAC.
Hope this helps.
gary g.
At 02:35 AM 10/9/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 12, 20 -- From gary-at-gaugler.com Mon Oct 9 12:35:58 2006 12, 20 -- Received: from qsmtp2.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 k99HZwTn009964 12, 20 -- for {microscopy-at-microscopy.com} ; Mon, 9 Oct 2006 12:35:58 -0500 12, 20 -- Received: (qmail 9350 invoked from network); 9 Oct 2006 10:35:57 -0700 12, 20 -- Received: by simscan 1.1.0 ppid: 9347, pid: 9348, t: 0.1485s 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 qsmtp2 with SMTP; 9 Oct 2006 10:35:57 -0700 12, 20 -- Message-Id: {7.0.1.0.2.20061009102243.025c3fd8-at-gaugler.com} 12, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 20 -- Date: Mon, 09 Oct 2006 10:35:59 -0700 12, 20 -- To: mmiralles-at-pi.ac.ae 12, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 20 -- Subject: Re: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 12, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 20 -- In-Reply-To: {200610090935.k999ZeJY015446-at-ns.microscopy.com} 12, 20 -- References: {200610090935.k999ZeJY015446-at-ns.microscopy.com} 12, 20 -- Mime-Version: 1.0 12, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
David, sorry for my late answer; I had been on holidays in Italy :-)))
The DSMs are very nice instruments but from what I know a little complicated in trouble-shooting. If you need some original service manuals I can try and copy these from a friend of mine.
Yes, definitely there should be more high-tech available at high-schools or some joint-ventures between high-schools and universities to share possibilities...
In a kind of way I am running out of "service-time" adopting more than the existing two schools, but - thinking of all those field-engineers out there from different manufacturers someone at FEI, HITACHI, JEOL or ZEISS NTS should think about doing some adoption of schools...
Modells for integration within the school system very largely depend on the teachers there (my experience) and your time available for coaching kids and teachers and doing updates sometimes a year. Get the machines known throughout the school, do some nice image and put it on floor walls....
----- Original Message ----- X-from: "David L. Jones" {dljones-at-bestweb.net} To: {stefan.diller-at-t-online.de} Sent: Thursday, September 28, 2006 6:45 PM
I was very interested in the several different ways people have developed for back-filling SEM specimen chambers with dry nitrogen gas. I think, however, that years ago we developed a method that is even simpler and less expensive than any of those described so far. This method is described in detail on page 65 of my book Vacuum Methods in Electron Microscopy (available from SPI Supplies, M. E. Taylor, Ladd, etc.)
What we did was to run a flexible hose (ordinary polyethylene tubing does just fine) from a stopper in the mouth of the Dewar flask of an EDS detector (you could alternatively connect to the bleed-off line on a liquid nitrogen storage tank) to the gas inlet on the SEM's specimen chamber. A large flexible plastic container, such as an inflatable plastic toy or beach ball, was connected to a T-junction in this line by means of a piece of soft, flexible surgical rubber tubing, and a straight, clean slit about 100 mm long was made in this tubing with a sharp razor blade or scalpel. This slit will normally stay closed tightly enough so that the nitrogen boiling off from the liquid nitrogen container will flow into the plastic ball. When the ball becomes full, the slit will open enough to prevent the system from becoming over pressurized, thus serving as a primitive pressure relief valve. When the inlet valve to the specimen chamber is opened, the plastic ball will collapse and the nitrogen will flow into the specimen chamber, but only under atmospheric pressure so there will be no danger of over pressurization. A small weight can be placed on the plastic ball to maintain a small flow of nitrogen after the system reaches atmospheric pressure, if desired. A beach ball about 0.5 m in diameter will hold enough nitrogen to fill most vacuum chambers several times, and it turns out to be lots of fun explaining to visitors why you have the ball connected to your electron microscope.
-- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-engin.umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
==============================Original Headers============================== 3, 14 -- From bigelow-at-engin.umich.edu Mon Oct 9 15:38:36 2006 3, 14 -- Received: from srvr20.engin.umich.edu (srvr20.engin.umich.edu [141.213.75.22]) 3, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k99KcaE0002154 3, 14 -- for {microscopy-at-microscopy.com} ; Mon, 9 Oct 2006 15:38:36 -0500 3, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 3, 14 -- by srvr20.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k99KcaVN017226 3, 14 -- for {microscopy-at-microscopy.com} ; Mon, 9 Oct 2006 16:38:36 -0400 (EDT) 3, 14 -- Mime-Version: 1.0 3, 14 -- Message-Id: {p06210202c1505bfe49ec-at-[141.212.131.221]} 3, 14 -- Date: Mon, 9 Oct 2006 16:38:35 -0400 3, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 3, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 3, 14 -- Subject: [Microscopy] RE: Venting Specimen Chambers 3, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================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 jonas-baltrusaitis-at-uiowa.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: jonas-baltrusaitis-at-uiowa.edu Name: Jonas Baltrusaitis
Organization: Central Microscopy Research Facility, Univ. of Iowa
Title-Subject: [Filtered] SEC from FESEM
Question: All,
Where can I purchase an old SEM specimen exchange chamber? I want to modify it and use as a quick load chamber on my existing Kratos AXIS Ultra XPS system.
RESEARCH ASSOCIATE 3 OR 4 (Microscopy Imaging Specialist) Department of Biological Sciences
The Socolofsky Microscopy Facility at LSU (http://www.biology.lsu.edu/facilities/micro_fac/) has state of the art light microscopy equipment, including a Leica TCS SP2 confocal system, deconvolution imaging system based on a Leica DM RXA2 upright, and a newly acquired Zeiss Lumar.V12 fluorescence stereomicroscope. Required Qualifications: (Research Associate 3): Bachelor's or equivalent degree in Biological Science or related field with three years relevant experience OR a Master's degree with one year experience. (Research Associate 4): Master's degree with two years relevant experience OR Ph.D. Experience in confocal light microscopy is REQUIRED for either level (RA3 or RA4). NOTE: This announcement is for ONE position at either RA3 or RA4 level, not for two positions.
Responsibilities: images acquisition and analysis, user training, and oversight of light microscopes and multi-user digital imaging equipment. An offer of employment is contingent on a satisfactory pre-employment background check. Application deadline is November 10, 2006, or until candidate is selected.
Submit letter of application and resume (including email address), and two letters of recommendation to:
Ms. Charyl Thompson Dept. of Biological Sciences 202 Life Sciences Bldg. Louisiana State University Ref: #014225 Baton Rouge, LA 70803. E-mail: cthomps-at-lsu.edu
LSU IS AN EQUAL OPPORTUNITY/EQUAL ACCESS EMPLOYER
_______________________________________________
David H. Burk, Ph.D. Socolofsky Microscopy Center Department of Biological Sciences 202 Life Sciences Building Louisiana State University Office: (225)578-8246 Fax: (225)578-2597 dburk-at-lsu.edu
==============================Original Headers============================== 10, 20 -- From dburk-at-lsu.edu Tue Oct 10 09:20:52 2006 10, 20 -- Received: from gate011.lsu.edu (gate011.ocs.lsu.edu [130.39.184.213]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9AEKpiG006232 10, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 10 Oct 2006 09:20:51 -0500 10, 20 -- Received: from smc30 ([130.39.115.181]) 10, 20 -- by gate011.lsu.edu (Lotus Domino Release 6.0.3) 10, 20 -- with ESMTP id 2006101009205099-1197 ; 10, 20 -- Tue, 10 Oct 2006 09:20:50 -0500 10, 20 -- From: "David Burk" {dburk-at-lsu.edu} 10, 20 -- To: {Microscopy-at-microscopy.com} 10, 20 -- Subject: Position Available - Microscopy Imaging Specialist 10, 20 -- Date: Tue, 10 Oct 2006 09:20:48 -0500 10, 20 -- Message-ID: {000c01c6ec77$483d0620$b5732782-at-biosc.lsu.edu} 10, 20 -- MIME-Version: 1.0 10, 20 -- X-Mailer: Microsoft Office Outlook 11 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 10, 20 -- Thread-Index: Acbsd0guMJVzZ0wWSPKoCUaPciQA8g== 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="us-ascii" ==============================End of - Headers==============================
I don't see Topaz much either. I replaced the Topaz unit several years ago and do not have the model number here any longer. It probably got replaced by a new one or the line shut down. What you need is a ferroresonant transformer like:
These are USA links but probably TSI or Sola have international outlets.
If you check the data on the links, these are probably what you seek rather than an UPS. If power is on all the time but dirty, the ferroresonant units are the ticket. They can be much cheaper than an UPS of same capacity. Plus, they easily can handle three phase input power if you have it.
gary g.
At 02:07 AM 10/10/2006, you wrote:
} Hi gary, } } Can you give me a more specific model of this Topaz stabilizer? I only } saw a single model at the web and it didn't post the technical details. } } Thanks again, } Melina } } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Monday, October 09, 2006 9:36 PM } To: Melina Miralles } Cc: MSA listserver } Subject: Re: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 } } If your concern is surges or sags in power supply, you } don't need an UPS. What you would need is a stabilizer } like Topaz makes. This is a magnetic resonance system } that adjusts output voltage to be at specified output. } It also includes input spike protection and EMI/RFI } removal. } } If you do want an UPS, and these are very nice to have, } there are several makers. I use two Liebert Nfinity } 8KVA units with redundant power and control modules. } SEM and its PC are on one UPS while EDS, EBSD and other } PCs and chiller and compressor are on another one. Each } UPS runs at about 25% total capacity and will run the } whole system for about 250 minutes if power fails--which } it usually does in winter and summer here in California. } These units regulate output voltage to within about 2% of } specified value. They are double conversion in that they } take in AC, convert it to DC and invert the DC to AC output. } When input AC fails, DC from the batteries is inverted to AC } output. Along with this is spike protection, sag protection } and EMI/RFI protection. } } These units work on 208-240VAC input and output similar } voltage. I take 240VAC in and output 240VAC and 120VAC. } } Hope this helps. } } gary g.
==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Tue Oct 10 18:20:24 2006 11, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9ANKOjQ028072 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 10 Oct 2006 18:20:24 -0500 11, 20 -- Received: (qmail 14657 invoked from network); 10 Oct 2006 16:17:22 -0700 11, 20 -- Received: by simscan 1.1.0 ppid: 14652, pid: 14654, t: 0.1668s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp4 with SMTP; 10 Oct 2006 16:17:22 -0700 11, 20 -- Message-Id: {7.0.1.0.2.20061010161328.0257cd40-at-gaugler.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 20 -- Date: Tue, 10 Oct 2006 16:20:25 -0700 11, 20 -- To: "Melina Miralles" {mmiralles-at-pi.ac.ae} 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: RE: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {D5603421C6303A46883F87E7968F285B01DEAA28-at-pi-exm.PI.AC.AE} 11, 20 -- References: {D5603421C6303A46883F87E7968F285B01DEAA28-at-pi-exm.PI.AC.AE} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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 robertoclark-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: robertoclark-at-gmail.com Name: Rob Clark
Organization: Sharp HealthCare
Title-Subject: [Filtered] Decacifying teeth
Question: Hi All,
I am debating whether or not to decalcify a root of a tooth in a formic acid/formaldehyde decal such as CalRite, or to go the slow route with EDTA/glut. The root will then be processed for TEM studies in hopes to identify microorganisms. No other studies are planned. From your experience will formic acid be the way to go or will it compromise my results?
Thank you so much in advance. Rob Clark E.M. Technologist Sharp HealthCare San Diego CA
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Email: ppq3k-at-yahoo.com Name: Peng Li
Organization: University of New Mecico
Title-Subject: [Filtered] Electron Microscopist position at Univ. of New Mexico
Question: Research Scientist 1/Electron Microscopist Department of Earth and Planetary Sciences, University of New Mexico
The University of New Mexico at UNM is seeking applicants for a Research Scientist 1/Electron Microscopist for the Transmission Electron Microscope Laboratory in the Department of Earth and Planetary Sciences. The laboratory is a state of the art multiuser facility that includes JEOL 2010 high resolution TEM and JEOL 2010F FASTEM FEG TEM/STEM with an Oxford ISIS EDS and GATAN GIF system, and sample preparation facilities including ion mills, ultramicrotome and carbon coaters. The Research Technologist will be responsible for the day to day running of the laboratory in close collaboration with the laboratory manager, including training and assisting student, staff and faculty users, purchasing of laboratory consumables, sample preparation, laboratory safety, billing and record keeping. The successful candidate will also be expected to maintain and troubleshoot laboratory and TEM equipment and supervise service engineers. Full details of the position and application procedure can be obtained at the website address: http://jobs.unm.edu/jobopenings.cfm. Closing date for the position is October 30, 2006. For additional information contact Professor Adrian Brearley (Tel: 505 277 4163, e-mail :brearley-at-unm.edu) or Dr Peng Li (Tel: 505 277 7536; e-mail: pengli-at-unm.edu).
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Title-Subject: [Filtered] An Free Kevex Light Element Detector
Question: We have an 15 year old Kevex detector (light elements, Be window) we are pitching. Is anyone interested? I still have the shipping box. You might have to pay for the shipping???? I'd have to ask TPTB.
Perhaps we could exchange our views about the optimal buffers to use during fixations for TEM. Cacodylate is a traditional buffer, but it is not really healthy and perhaps it is time to update our knowledge. I have read that PIPES offered the lowest extraction but on the downside a less effecient buffering. Phosphate buffer does not allow the use of Ca and Mg, which could help stabilize some structures. HEPES buffer...well I don't know the downside, perhaps it offers only advantages ;-)but it is used pretty scarcely by the electron microscopists. Some other parameters can be taken into account: the storage, bench (or fridge) life, morphology {} immunology, plants {} animals cells and so on.
Lets talk about salts!
Personally I use Cacodylate for classical fixation of cells for morphology because it has also been used in the laboratory but I am eager to change. There is certainly some room for improvements in the morphology (especially of the membranes). HEPES seems to be a good candidate.
Stephane
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Oct 11 03:09:36 2006 6, 18 -- Received: from web37409.mail.mud.yahoo.com (web37409.mail.mud.yahoo.com [209.191.91.141]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9B89aiA016895 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 03:09:36 -0500 6, 18 -- Received: (qmail 28241 invoked by uid 60001); 11 Oct 2006 08:09:36 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 18 -- b=ChPAJbr1a2yJNpAafId35uxgkFOweiyNadUClf3kn/3vCzkXkmVSq83kMSU39wM/SA9OE32fVdMpo7cVyn14pAUg5FAqfmoj/NjpDIBgezrja3Sg7yKHXl4a7DvEOWxRqWDD+BDfBGD/GIFHMAZw8Q9GrzBsnlVdSy1r3b2snjk= ; 6, 18 -- Message-ID: {20061011080936.28239.qmail-at-web37409.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37409.mail.mud.yahoo.com via HTTP; Wed, 11 Oct 2006 01:09:36 PDT 6, 18 -- Date: Wed, 11 Oct 2006 01:09:36 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: debating about buffers for TEM fixation 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- MIME-Version: 1.0 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 6, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (winnie.wino-at-gmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 11, 2006 at 04:28:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both winnie.wino-at-gmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: winnie.wino-at-gmail.com Name: Winnie
Organization: NUS
Education: Undergraduate College
Location: Singapore
Title: Sample preparation
Question: Dear Microscopist, I have a few specimens (Charcoal and virgin wood) of which the sizes are 100x100x50(mm) and 30x30x100(mm). What should I prepare for those specimens before bringing them to the Lab for SEM scanning? Can the SEM machine's chamber accomodate such specimens? Should I reduce the size? Thank you and Best Regards. Winnie
Buffers have been on my mind lately too. In this laboratory, we have switched our buffer system over to Dulbecco's DMEM cell culture medium. In large part this is a matter of convenience. We do quite a few immunocytochemistry experiments, labeling culture cell membranes with a variety of antibodies, and we dilute these antibodies and subsequent colloidal gold conjugates with DMEM. The cells remain viable during incubations in antibody and secondary conjugates. It seemed reasonable to us to continue with DMEM during fixation, and we therefore dilute our cacodylate buffered 3%glut/3%paraformaldehyde stock solution with DMEM and also our OsO4 in DMEM. We have been pleased with the quality of the fixation and now we also use DMEM for buffering fixatives of tissues with equally pleasing results. But I have to admit to a nagging feeling that I am breaking some kind of microscopy law and would appreciate comments.
Thanks,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Wednesday, October 11, 2006 1:15 AM To: drk-at-shcc.org
Wealthy colleagues (yes: knowledge is a wealth!),
Perhaps we could exchange our views about the optimal buffers to use during fixations for TEM. Cacodylate is a traditional buffer, but it is not really healthy and perhaps it is time to update our knowledge. I have read that PIPES offered the lowest extraction but on the downside a less effecient buffering. Phosphate buffer does not allow the use of Ca and Mg, which could help stabilize some structures. HEPES buffer...well I don't know the downside, perhaps it offers only advantages ;-)but it is used pretty scarcely by the electron microscopists. Some other parameters can be taken into account: the storage, bench (or fridge) life, morphology {} immunology, plants {} animals cells and so on.
Lets talk about salts!
Personally I use Cacodylate for classical fixation of cells for morphology because it has also been used in the laboratory but I am eager to change. There is certainly some room for improvements in the morphology (especially of the membranes). HEPES seems to be a good candidate.
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Oct 11 03:09:36 2006 6, 18 -- Received: from web37409.mail.mud.yahoo.com (web37409.mail.mud.yahoo.com [209.191.91.141]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9B89aiA016895 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 03:09:36 -0500 6, 18 -- Received: (qmail 28241 invoked by uid 60001); 11 Oct 2006 08:09:36 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content -Transfer-Encoding; 6, 18 -- b=ChPAJbr1a2yJNpAafId35uxgkFOweiyNadUClf3kn/3vCzkXkmVSq83kMSU39wM/SA9OE32fVd Mpo7cVyn14pAUg5FAqfmoj/NjpDIBgezrja3Sg7yKHXl4a7DvEOWxRqWDD+BDfBGD/GIFHMAZw8Q 9GrzBsnlVdSy1r3b2snjk= ; 6, 18 -- Message-ID: {20061011080936.28239.qmail-at-web37409.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37409.mail.mud.yahoo.com via HTTP; Wed, 11 Oct 2006 01:09:36 PDT 6, 18 -- Date: Wed, 11 Oct 2006 01:09:36 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: debating about buffers for TEM fixation 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- MIME-Version: 1.0 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 6, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 17, 21 -- From drk-at-shcc.org Wed Oct 11 10:19:56 2006 17, 21 -- Received: from SUNNIE.shcc.org (mail.shcc.org [64.213.211.202]) 17, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BFJtsx013068 17, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 10:19:55 -0500 17, 21 -- Received: from DRK2 ([64.213.211.62]) by shcc.org (PMDF V6.3 #31278) 17, 21 -- with ESMTP id {0J6Z00E0V8V2X5-at-shcc.org} for Microscopy-at-Microscopy.Com; Wed, 17, 21 -- 11 Oct 2006 08:11:26 -0700 (PDT) 17, 21 -- Date: Wed, 11 Oct 2006 08:19:49 -0700 17, 21 -- From: Doug Keene {drk-at-shcc.org} 17, 21 -- Subject: RE: [Microscopy] debating about buffers for TEM fixation 17, 21 -- In-reply-to: {200610110814.k9B8EqC0024591-at-ns.microscopy.com} 17, 21 -- To: Microscopy-at-Microscopy.Com 17, 21 -- Reply-to: drk-at-shcc.org 17, 21 -- Message-id: {0J6Z00E0W8V2X5-at-shcc.org} 17, 21 -- Organization: Shriners Hospitals for Children 17, 21 -- MIME-version: 1.0 17, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 17, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 17, 21 -- Content-type: text/plain; charset=us-ascii 17, 21 -- Content-transfer-encoding: 7bit 17, 21 -- Thread-Index: AcbtDChpSOQbSAYeQd2UYXPEktNGrQAOl+9g ==============================End of - Headers==============================
For 40 years we have used the physiological or experimental buffer solution, often with MOPS as the pH buffer, as our main vehicle for primary fixatives composed with aldehydes, aldehyde-tannic-acid, or tannic acid alone, because we believe this offers the best hope for preserving native structure of various muscle structural states (but also effective with other cells and tissues). "Physiological" mostly implies proper ions: for extracellular salines (Na+ at ~100-145 mM; K+ absent or below 2 mM; Mg++ and Ca++ at 1-4 mM); or intracellular rest-state salines (EGTA and no inadvertent Ca++; Mg at ~5-7 mM; K+ (or Na+) up to 150 mM if desired; chloride, propionate, acetate, or methane-sulfonate as anion).
So for intact frog leg muscle (or insect flight muscle!) we used frog Ringer solution, for mammalian muscle we used mammalian Ringer, with either phosphate or MOPS buffer, adjusting pH with a "dirty" (fixative only) pH electrode to discover the amount of alkali or acid needed to restore original pH after adding various fixative at various concentrations. The presence of up to 3 mM calcium and up to 10 mm magnesium, as dictated by various buffers, was essential in Ringer recipes, and presented no problem. We were happy because the fiber x-ray diffraction pattern and the EM itself showed these fixatives to give good preservation of various distinct pre-fixation native structural states, and i think this was not so for our brief long-ago trials of phosphate or s-collidine buffer. We usually ignored adjustments to osmolarity, but if tried, was always done by adding the osmolytge to the formulation, sucrose top Ringer, 500,000 MW dextran to intracellular buffer.
For secondary fixation, buffer choice need not be physiological. For aldehyde-only or aldehyde-TA primary fixation, post-fixing in OsO4 has routinely employed a Maupin-Pollard good-for-actin buffer (0.1M phosphate, pH 6, with 10 mM MgCl2, used ice cold).
Most of our work has been with chemically demembranated ("skinned"; by detergent-glycerol) muscle of rabbit, frog or insect, so we chose various composition of intracellular buffer to keep the organelles happy and in the desired functional state of the muscle machinery (rigor, Mg-ATP relaxed, Mg-ATP-Ca activating, or modified with nucleotide analogs (like AMPPNP) phosphate analogs (like vanadate). We have for 30 years used nothing but 20 mM MOPS buffer (relatively inexpensive, and good for pH 6.5-7.5; we prefer 6.8 for insect) for both the physiological and fixative versions of our solutions, typically with 5 mM each of additives like MgCl2, EGTA, Ca-EGTA, ATP, Na azide (stops microbial growth, arrest ATPase consumption by mitochondria). For years we strictly used potassium salts in preference to sodium to honor the cell's intracellular habit, but lately have slipped away from this dogma without bad results, drifting to more frequent use of sodium (as hydroxide to adjust pH of EGTA, ATP, MOPS, etc. Sometimes we use salt to approximate intracellular ionic strength, typically using KCl (now NaCl) at 100 or 150 ml, but we often use no extra salt at all, because its presence or absence has so far made little or no difference to the fiber x-ray diffraction patterns which are our gold standard for native and preserved structure.
We have always steered clear of Tris because glutaraldehyde reacts with it, changing pH (?? and what else?), and because Tris-buffered pH alters with changing temperature.
0.2% tannic acid alone in the physiological (intracellular type) buffer works well on skinned cells to fix all but soluble components. It is blocked or complexed so it becomes unavailable for fixation when Triton X-100 or PVP are included in the same solution. TA-fix should be followed after rinse-out by uranyl acetate (typically in DI water) or OsO4 (in Maupin-Pollard good-for-actin buffer) as a secondary fixative.
-mike reedy-
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************************ 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
This discussion takes me back to my doctoral research, in which I was concerned about the morphology of dissociated chick retina cells as they reassociated to form tissue-like aggregates. (Develop. Biol. 23:36-61, 1970(!)).
As part of that study, we fixed suspensions of cells in various concentrations of phosphate buffer, and measured the cell size distribution in the population with a Coulter Counter. We then chose to fix in 0.08 molar phosphate with 2.5% glutaraldehyde since that distribution matched the result with living cells.
It is clear that one can't just calculate osmotic strength for these systems, since the fixative alters the membrane behavior in a non- linear way during the fixation process.
Joel
-- Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
==============================Original Headers============================== 9, 27 -- From jbs-at-temple.edu Wed Oct 11 11:03:44 2006 9, 27 -- Received: from po-smtp1.temple.edu (po-smtp1.temple.edu [155.247.166.195]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BG3gbR002357 9, 27 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 11:03:43 -0500 9, 27 -- Received: from [155.247.98.40] (jbs.bio.temple.edu [155.247.98.40]) 9, 27 -- by po-smtp1.temple.edu (MOS 3.8.2-GA) 9, 27 -- with ESMTP id FXB69399 (AUTH jbs); 9, 27 -- Wed, 11 Oct 2006 12:03:40 -0400 (EDT) 9, 27 -- From: "Joel Sheffield" {jbs-at-temple.edu} 9, 27 -- To: microscopy-at-microscopy.com 9, 27 -- Date: Wed, 11 Oct 2006 12:01:47 -0400 9, 27 -- MIME-Version: 1.0 9, 27 -- Subject: Re: [Microscopy] Re: debating about buffers for TEM fixation 9, 27 -- Reply-to: jbs-at-temple.edu 9, 27 -- Message-ID: {452CDD2B.17834.904B15E2-at-jbs.temple.edu} 9, 27 -- Priority: normal 9, 27 -- In-reply-to: {200610111534.k9BFYeEf023954-at-ns.microscopy.com} 9, 27 -- X-mailer: Pegasus Mail for Windows (4.30 public beta 1) 9, 27 -- Content-type: text/plain; charset=US-ASCII 9, 27 -- Content-transfer-encoding: 7BIT 9, 27 -- Content-description: Mail message body 9, 27 -- X-Junkmail-Status: score=10/50, host=po-smtp1.temple.edu 9, 27 -- X-Junkmail-SD-Raw: score=unknown, 9, 27 -- refid=str=0001.0A09020A.452D1597.00CD,ss=1,fgs=0, 9, 27 -- ip=155.247.98.40, 9, 27 -- so=2006-09-22 03:48:54, 9, 27 -- dmn=5.1.5/2006-01-31 ==============================End of - Headers==============================
Dear Winnie, The biggest problem with wood and, to a lesser extent charcoal, is that it is very porous, has water in its structure and will out-gas a lot of water vapour. Fresh concrete has the same problem. I have had to cut my wood samples down to less than 10 mm. cube in order to get the SEM to pump down to high vacuum. If you are looking at the wood samples in a variable-pressure SEM it should not be such a big problem. Most modern SEMs, unless they are very high resolution, will look at a 100 mm square sample, it is the pumping capability that might be a problem. Regards,
Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca -----Original Message----- X-from: winnie.wino-at-gmail.com [mailto:winnie.wino-at-gmail.com] Sent: Wednesday, October 11, 2006 6:20 AM To: mager-at-interchange.ubc.ca
This Question was submitted to Ask-A-Microscopist by (winnie.wino-at-gmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 11, 2006 at 04:28:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both winnie.wino-at-gmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: winnie.wino-at-gmail.com Name: Winnie
Organization: NUS
Education: Undergraduate College
Location: Singapore
Title: Sample preparation
Question: Dear Microscopist, I have a few specimens (Charcoal and virgin wood) of which the sizes are 100x100x50(mm) and 30x30x100(mm). What should I prepare for those specimens before bringing them to the Lab for SEM scanning? Can the SEM machine's chamber accomodate such specimens? Should I reduce the size? Thank you and Best Regards. Winnie
I have used serum free media for many cell culture fixations. I think it works fine. After fix I go into CaCo so that I do not have any interactions with my TC media and any post-fix steps. David
On Oct 11, 2006, at 8:25 AM, drk-at-shcc.org 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 } ---------------------------------------------------------------------- } ------ } } Fellow Microscopists, } } Buffers have been on my mind lately too. In this laboratory, we have } switched our buffer system over to Dulbecco's DMEM cell culture } medium. In } large part this is a matter of convenience. We do quite a few } immunocytochemistry experiments, labeling culture cell membranes } with a } variety of antibodies, and we dilute these antibodies and subsequent } colloidal gold conjugates with DMEM. The cells remain viable during } incubations in antibody and secondary conjugates. It seemed } reasonable to } us to continue with DMEM during fixation, and we therefore dilute our } cacodylate buffered 3%glut/3%paraformaldehyde stock solution with } DMEM and } also our OsO4 in DMEM. We have been pleased with the quality of the } fixation } and now we also use DMEM for buffering fixatives of tissues with } equally } pleasing results. But I have to admit to a nagging feeling that I am } breaking some kind of microscopy law and would appreciate comments. } } Thanks, } } Doug } } Douglas R. Keene } Assistant Investigator } Micro-Imaging Center } Shriners Hospital for Children } 3102 S.W. Sam Jackson Park Road } Portland, Oregon 97239 } 503-221-3434 } drk-at-shcc.org } } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Wednesday, October 11, 2006 1:15 AM } To: drk-at-shcc.org } Subject: [Microscopy] debating about buffers for TEM fixation } } } } } ---------------------------------------------------------------------- } ------ } 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 } ---------------------------------------------------------------------- } ------ } } Wealthy colleagues (yes: knowledge is a wealth!), } } Perhaps we could exchange our views about the optimal } buffers to use during fixations for TEM. } Cacodylate is a traditional buffer, but it is not } really healthy and perhaps it is time to update our } knowledge. I have read that PIPES offered the lowest } extraction but on the downside a less effecient } buffering. Phosphate buffer does not allow the use of } Ca and Mg, which could help stabilize some structures. } HEPES buffer...well I don't know the downside, perhaps } it offers only advantages ;-)but it is used pretty } scarcely by the electron microscopists. } Some other parameters can be taken into account: the } storage, bench (or fridge) life, } morphology {} immunology, plants {} animals cells and so } on. } } Lets talk about salts! } } Personally I use Cacodylate for classical fixation of } cells for morphology because it has also been used in } the laboratory but I am eager to change. There is } certainly some room for improvements in the morphology } (especially of the membranes). HEPES seems to be a } good candidate. } } Stephane } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Wed Oct 11 03:09:36 2006 } 6, 18 -- Received: from web37409.mail.mud.yahoo.com } (web37409.mail.mud.yahoo.com [209.191.91.141]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k9B89aiA016895 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 03:09:36 } -0500 } 6, 18 -- Received: (qmail 28241 invoked by uid 60001); 11 Oct 2006 } 08:09:36 } -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content- } Type:Content } -Transfer-Encoding; } 6, 18 -- } b=ChPAJbr1a2yJNpAafId35uxgkFOweiyNadUClf3kn/3vCzkXkmVSq83kMSU39wM/ } SA9OE32fVd } Mpo7cVyn14pAUg5FAqfmoj/NjpDIBgezrja3Sg7yKHXl4a7DvEOWxRqWDD+BDfBGD/ } GIFHMAZw8Q } 9GrzBsnlVdSy1r3b2snjk= ; } 6, 18 -- Message-ID: } {20061011080936.28239.qmail-at-web37409.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37409.mail.mud.yahoo.com via } HTTP; Wed, 11 Oct 2006 01:09:36 PDT } 6, 18 -- Date: Wed, 11 Oct 2006 01:09:36 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: debating about buffers for TEM fixation } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- MIME-Version: 1.0 } 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 6, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 17, 21 -- From drk-at-shcc.org Wed Oct 11 10:19:56 2006 } 17, 21 -- Received: from SUNNIE.shcc.org (mail.shcc.org } [64.213.211.202]) } 17, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k9BFJtsx013068 } 17, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 } 10:19:55 -0500 } 17, 21 -- Received: from DRK2 ([64.213.211.62]) by shcc.org (PMDF } V6.3 #31278) } 17, 21 -- with ESMTP id {0J6Z00E0V8V2X5-at-shcc.org} for } Microscopy-at-Microscopy.Com; Wed, } 17, 21 -- 11 Oct 2006 08:11:26 -0700 (PDT) } 17, 21 -- Date: Wed, 11 Oct 2006 08:19:49 -0700 } 17, 21 -- From: Doug Keene {drk-at-shcc.org} } 17, 21 -- Subject: RE: [Microscopy] debating about buffers for TEM } fixation } 17, 21 -- In-reply-to: {200610110814.k9B8EqC0024591-at-ns.microscopy.com} } 17, 21 -- To: Microscopy-at-Microscopy.Com } 17, 21 -- Reply-to: drk-at-shcc.org } 17, 21 -- Message-id: {0J6Z00E0W8V2X5-at-shcc.org} } 17, 21 -- Organization: Shriners Hospitals for Children } 17, 21 -- MIME-version: 1.0 } 17, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 } 17, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 } 17, 21 -- Content-type: text/plain; charset=us-ascii } 17, 21 -- Content-transfer-encoding: 7bit } 17, 21 -- Thread-Index: AcbtDChpSOQbSAYeQd2UYXPEktNGrQAOl+9g } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Wed Oct 11 11:47:04 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BGl47S024189 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 11:47:04 -0500 5, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 05AB411299C2 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 09:46:48 -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 C9F4E1129080 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 09:40:16 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200610111525.k9BFPInw018425-at-ns.microscopy.com} 5, 22 -- References: {200610111525.k9BFPInw018425-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {699624AD-10B7-4A90-BC11-A49FBBD99E1D-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] RE: debating about buffers for TEM fixation 5, 22 -- Date: Wed, 11 Oct 2006 09:40:14 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Greetings, Mike Reedy mentioned that he uses MOPS for a buffer at pH 7 for fixations. I have always used PIPES. Has anyone ever compared the performance of these two Good buffers around neutral pH?? Are there reasons to choose one vs the other?
On Oct 11, 2006, at 10:04 AM, baskin-at-bio.umass.edu wrote:
} Mike Reedy mentioned that he uses MOPS for a buffer at pH 7 } for fixations. I have always used PIPES. Has anyone ever compared the } performance of these two Good buffers around neutral pH?? Are there } reasons to choose one vs the other? } Dear Tobias, The ability of a buffer to maintain pH depends on how many groups there are on the molecule that can take up or release H+ and the pK of each of these groups. For most buffers there are one or a few of these groups that have different pKs; e.g. PO4--- with pKs of ~12, ~7, and ~3. I don't know offhand what pKs are available for either MOPS or PIPES, but the closer a pK is to the pH you want, the better the buffer will be at maintaining that pH. The reason for this is easily seen from the mass action law, which can be written
K = [A][H]/[AH]
from which it can be seen that for [H] = K, [A] = [AH], and if [H] = K, then pH = pK. When the protonated and unprotonated species are at equal concentration, the addition of a small amount of acid or base will change the [A]/[AH] ratio less than if they are at unequal concentration. Therefore, the performance of either Good buffer will equal that of the other one if the desired pH is the same amount away from either pK. Another reason to choose one over another is if one of them affects the process you are trying to perform. It may be that one will react with your fixative--which I do not think is the case for MOPS or PIPES and either glut or OsO4. 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============================== 6, 22 -- From tivol-at-caltech.edu Wed Oct 11 14:52:43 2006 6, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BJqh5u016670 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 14:52:43 -0500 6, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 6, 22 -- by water-ox-postvirus (Postfix) with ESMTP id CEFFF2EF2E 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 12:52:42 -0700 (PDT) 6, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 6, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 64B9C109AED 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 12:52:41 -0700 (PDT) 6, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 6, 22 -- In-Reply-To: {200610111704.k9BH4Ypv002703-at-ns.microscopy.com} 6, 22 -- References: {200610111704.k9BH4Ypv002703-at-ns.microscopy.com} 6, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 6, 22 -- Message-Id: {4cb53a98ada174cee4b63e632eab0ca7-at-caltech.edu} 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 6, 22 -- Subject: Re: [Microscopy] MOPS vs PIPES 6, 22 -- Date: Wed, 11 Oct 2006 12:56:56 -0700 6, 22 -- To: microscopy-at-msa.microscopy.com 6, 22 -- X-Mailer: Apple Mail (2.624) 6, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
What is your experience with the film camera? How many successful film cassettes do you average between camera jams? Please quote for each TEM model (100CX, 2000FX, 3100, etc).
Thanks, Roseann
Roseann Csencsits, PhD. Donner TEM Facility Manager Lawrence Berkeley Lab 510-486-4548
==============================Original Headers============================== 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} 7, 21 -- Subject: JEOL camera question 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- X-Mailer: Apple Mail (2.752.3) 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 7, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I have been reading this thread with interest in the hope of learning something new. The theory of buffers as explained by Bill Tivol, doesn't seem to be useful when applied to electron microscopy. The reason is that we judge the effects of different buffers on their effect on morphology (as Mike Reedy subtly puts it in his message), not on the final pH of the specimens.
Judged on buffering capacity alone, sodium cacodylate would fail miserably as a buffer. The Good buffers (PIPES, HEPES etc) appear to be excellent buffers for keeping solutions at a stable pH. However, even if PIPES or HEPES is used to buffer aldehyde, when the solution encounters biological material there is a sudden drop in pH that is not affected at all by the buffer in use. Anyone who has fixed cell cultures in the presence of medium containing pH indicator can attest to the rapid color change that occurs.
It is worth noting that the pH to which buffers are adjusted, and the pH at which they are most effective, is not always 7.2 or 7.4. For example, the use of PIPES at a pH higher than 6.8 will greatly reduce its buffering capacity.
I liked the idea of comparing the sizes of fixed cells with living cells to determine which concentration of buffer is "best" to use. There was no mention of what the intracellular morphology looked like and if it was acceptable. I am sure that the best concentration will differ between cell types. The great disadvantage of this method for determining "correct' buffer concentrations is that most of our work involves tissues composed of many different cell types that are all attached to each other.
Another issue touched on by Mike Reedy is the state of aldehyde-fixed biological material. Is is really osmotically inactive as assumed (e.g. "...buffer choice need not be physiological.")? Could aqueous solutions of osmium tetroxide be used in post-fixation steps? Are aldehyde-fixed membranes osmotically inactive?
Our final evaluation of buffers will depend on what we are looking at, or looking for, in our specimens. Sometimes we need to extract lots of cytoplasmic components to give high-contrast images. Other times we need to retain as many proteins as possible so that we can perform immunocytochemical labeling. Other important considerations for immunolabeling are the need to retain all antigens at their "natural" site, yet enable complete antibody accessibility to antigens.
For the record, our routine fixation protocols involve glutaraldehyde and sodium cacodylate for morphology, and double-strength aldehyde (formaldehyde or glutaraldehyde, or mixtures of both) in double strength HEPES buffer for immunocytochemistry. The double-strength fixative is added to culture medium, ringer or other physiological medium. There routine fixatives work well for us, but we do not work with sharks, molluscs or plants.
Any EM laboratory involved in many projects may find that one fixation protocol is insufficient for all users. Instead, different protocols involving different buffers gradually evolve in the lab to cope with each specimen as needed. What we need is a good understanding of how buffers and cross-linking chemicals affect the specimens we apply them to.
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
The ability of a buffer to maintain pH depends on how many groups there are on the molecule that can take up or release H+ and the pK of each of these groups. For most buffers there are one or a few of these groups that have different pKs; e.g. PO4--- with pKs of ~12, ~7, and ~3. I don't know offhand what pKs are available for either MOPS or PIPES, but the closer a pK is to the pH you want, the better the buffer will be at maintaining that pH. The reason for this is easily seen from the mass action law, which can be written
K = [A][H]/[AH]
from which it can be seen that for [H] = K, [A] = [AH], and if [H] = K, then pH = pK. When the protonated and unprotonated species are at equal concentration, the addition of a small amount of acid or base will change the [A]/[AH] ratio less than if they are at unequal concentration. Therefore, the performance of either Good buffer will equal that of the other one if the desired pH is the same amount away from either pK. Another reason to choose one over another is if one of them affects the process you are trying to perform. It may be that one will react with your fixative--which I do not think is the case for MOPS or PIPES and either glut or OsO4. Yours, Bill Tivol, PhD
For 40 years we have used the physiological or experimental buffer solution, often with MOPS as the pH buffer, as our main vehicle for primary fixatives composed with aldehydes, aldehyde-tannic-acid, or tannic acid alone, because we believe this offers the best hope for preserving native structure of various muscle structural states (but also effective with other cells and tissues). "Physiological" mostly implies proper ions: for extracellular salines (Na+ at ~100-145 mM; K+ absent or below 2 mM; Mg++ and Ca++ at 1-4 mM); or intracellular rest-state salines (EGTA and no inadvertent Ca++; Mg at ~5-7 mM; K+ (or Na+) up to 150 mM if desired; chloride, propionate, acetate, or methane-sulfonate as anion).
So for intact frog leg muscle (or insect flight muscle!) we used frog Ringer solution, for mammalian muscle we used mammalian Ringer, with either phosphate or MOPS buffer, adjusting pH with a "dirty" (fixative only) pH electrode to discover the amount of alkali or acid needed to restore original pH after adding various fixative at various concentrations. The presence of up to 3 mM calcium and up to 10 mm magnesium, as dictated by various buffers, was essential in Ringer recipes, and presented no problem. We were happy because the fiber x-ray diffraction pattern and the EM itself showed these fixatives to give good preservation of various distinct pre-fixation native structural states, and i think this was not so for our brief long-ago trials of phosphate or s-collidine buffer. We usually ignored adjustments to osmolarity, but if tried, was always done by adding the osmolytge to the formulation, sucrose top Ringer, 500,000 MW dextran to intracellular buffer.
For secondary fixation, buffer choice need not be physiological. For aldehyde-only or aldehyde-TA primary fixation, post-fixing in OsO4 has routinely employed a Maupin-Pollard good-for-actin buffer (0.1M phosphate, pH 6, with 10 mM MgCl2, used ice cold).
Most of our work has been with chemically demembranated ("skinned"; by detergent-glycerol) muscle of rabbit, frog or insect, so we chose various composition of intracellular buffer to keep the organelles happy and in the desired functional state of the muscle machinery (rigor, Mg-ATP relaxed, Mg-ATP-Ca activating, or modified with nucleotide analogs (like AMPPNP) phosphate analogs (like vanadate). We have for 30 years used nothing but 20 mM MOPS buffer (relatively inexpensive, and good for pH 6.5-7.5; we prefer 6.8 for insect) for both the physiological and fixative versions of our solutions, typically with 5 mM each of additives like MgCl2, EGTA, Ca-EGTA, ATP, Na azide (stops microbial growth, arrest ATPase consumption by mitochondria). For years we strictly used potassium salts in preference to sodium to honor the cell's intracellular habit, but lately have slipped away from this dogma without bad results, drifting to more frequent use of sodium (as hydroxide to adjust pH of EGTA, ATP, MOPS, etc. Sometimes we use salt to approximate intracellular ionic strength, typically using KCl (now NaCl) at 100 or 150 ml, but we often use no extra salt at all, because its presence or absence has so far made little or no difference to the fiber x-ray diffraction patterns which are our gold standard for native and preserved structure.
We have always steered clear of Tris because glutaraldehyde reacts with it, changing pH (?? and what else?), and because Tris-buffered pH alters with changing temperature.
0.2% tannic acid alone in the physiological (intracellular type) buffer works well on skinned cells to fix all but soluble components. It is blocked or complexed so it becomes unavailable for fixation when Triton X-100 or PVP are included in the same solution. TA-fix should be followed after rinse-out by uranyl acetate (typically in DI water) or OsO4 (in Maupin-Pollard good-for-actin buffer) as a secondary fixative.
-mike reedy-
==============================Original Headers============================== 31, 18 -- From PWebster-at-hei.org Wed Oct 11 16:18:24 2006 31, 18 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 31, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BLIOUn006955 31, 18 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 16:18:24 -0500 31, 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 ; 31, 18 -- Wed, 11 Oct 2006 21:18:23 +0000 31, 18 -- User-Agent: Microsoft-Entourage/11.2.5.060620 31, 18 -- Date: Wed, 11 Oct 2006 14:18:23 -0700 31, 18 -- Subject: Re: debating about buffers for TEM fixation 31, 18 -- From: "Webster, Paul" {PWebster-at-hei.org} 31, 18 -- To: {microscopy-at-msa.microscopy.com} 31, 18 -- Message-ID: {C152ADAF.C5ED%PWebster-at-hei.org} 31, 18 -- Thread-Topic: debating about buffers for TEM fixation 31, 18 -- Thread-Index: AcbteshdByXfOlluEdua3wANk7Zh7g== 31, 18 -- Mime-version: 1.0 31, 18 -- Content-type: text/plain; 31, 18 -- charset="US-ASCII" 31, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
Roseann; My experience is that it is more a function of care and attention to detail of loading film into the cassettes than anything else.
John Mardinly Intel Corporation
Disclaimer: This is the opinion of this individual, not the opinion of Intel Corporation.
-----Original Message----- X-from: rcsencsits-at-lbl.gov [mailto:rcsencsits-at-lbl.gov] Sent: Wednesday, October 11, 2006 2:07 PM To: Mardinly, John
Dear JEOL film camera users:
What is your experience with the film camera? How many successful film cassettes do you average between camera jams? Please quote for each TEM model (100CX, 2000FX, 3100, etc).
Thanks, Roseann
Roseann Csencsits, PhD. Donner TEM Facility Manager Lawrence Berkeley Lab 510-486-4548
==============================Original Headers============================== 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} 7, 21 -- Subject: JEOL camera question 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- X-Mailer: Apple Mail (2.752.3) 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 7, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
==============================Original Headers============================== 16, 31 -- From john.mardinly-at-intel.com Wed Oct 11 19:48:10 2006 16, 31 -- Received: from mga03.intel.com (mga03.intel.com [143.182.124.21]) 16, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C0m9Ma021539 16, 31 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 19:48:10 -0500 16, 31 -- Received: from azsmga001.ch.intel.com ([10.2.17.19]) 16, 31 -- by mga03.intel.com with ESMTP; 11 Oct 2006 17:48:07 -0700 16, 31 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 16, 31 -- by azsmga001.ch.intel.com with ESMTP; 11 Oct 2006 17:48:07 -0700 16, 31 -- X-ExtLoop1: 1 16, 31 -- X-IronPort-AV: i="4.09,297,1157353200"; 16, 31 -- d="scan'208"; a="129889456:sNHT24905965" 16, 31 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 16, 31 -- Wed, 11 Oct 2006 17:48:06 -0700 16, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 31 -- Content-class: urn:content-classes:message 16, 31 -- MIME-Version: 1.0 16, 31 -- Content-Type: text/plain; 16, 31 -- charset="us-ascii" 16, 31 -- Subject: RE: [Microscopy] JEOL camera question 16, 31 -- Date: Wed, 11 Oct 2006 17:48:05 -0700 16, 31 -- Message-ID: {1DF4C4D62339DB4C9C98DF04213995B60170A5A2-at-scsmsx413.amr.corp.intel.com} 16, 31 -- X-MS-Has-Attach: 16, 31 -- X-MS-TNEF-Correlator: 16, 31 -- Thread-Topic: [Microscopy] JEOL camera question 16, 31 -- Thread-Index: AcbteTQlou9AYV+TSiKWxqGREJ+P3AAHfgrA 16, 31 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 16, 31 -- To: {rcsencsits-at-lbl.gov} 16, 31 -- Cc: {Microscopy-at-msa.microscopy.com} 16, 31 -- X-OriginalArrivalTime: 12 Oct 2006 00:48:06.0868 (UTC) FILETIME=[14EF5940:01C6ED98] 16, 31 -- Content-Transfer-Encoding: 8bit 16, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9C0m9Ma021539 ==============================End of - Headers==============================
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Email: kychung2-at-wisc.edu Name: Ka Young Chung
Organization: University of Wisconsin-Madison
Title-Subject: [Filtered] Fixation problem
Question: I am working with cardiac myocytes. To perform immunofluorescence, I have permeabilized cells with saponin without fixation, and could get nice signal. The signal was blocked when I used blocking peptide (original immunogen for primary antibody). To look at the localization pattern more in detail, I want to do electron microscopy. In this case, I need to fix the cells. To check whether fixation affects antibody interaction, I fixed cells with 4% paraformaldehyde and 0.1% glutaraldehyde, permeabilized cells with saponin, and then performed immunofluorecence. I saw a permeabilization problem with this experiment so that I tried 0.1% triton instead of saponin. Still, I am not getting the same staining pattern that I got from unfixed cells. How can I solve this problem? Can I just try EM?
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Question: I am searching for info.relating to an antique bausch&lomb metallograph. If anyone has related info. it would be so very much appreciated. I can email photos of the metallogragh to anyone who might have the knowledge to enlighten me about this. It is in good working condition,with many accesories ,assorted lenses and interchangable cameras. It is a large free standind, two pedestal, piece of equiptment, with camera like devices at each end. Stamp into it is the following info.
BAUSCH & LOMB OPTICAL CO. ROCHESTER, NY 01348
one div. =0.002mm Serial No.-6097 US Pat. 1798634 2074106
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Email: mdelann1-at-jhmi.edu Name: Michael Delannoy
Organization: Assistant Director Imaging Facility JHSM
Title-Subject: [Filtered] chillers
Question: Hello I need a few companies who would sell used/ refurbished water chillers/recirculators. I need a supply for my LEO 1530 FESEM Does anyone know the heat capaciy required? Does anyone know the Neslabs website or contact info? Thank You M Delannoy 410 955-1365
We have a Hitachi H-600 TEM, 1983. For the past year we've been operating with a faulty CRT data display unit. Each morning the display would come on for about ten minutes, then it would become unstable and disappear for the rest of the day. Finally it appears to have died completely which makes operating the TEM tricky. Our in-house electrician has had several attempts at fixing the problem but without success. The microscope itself is completely functional so operations such as column alignment, resetting number of unexposed negatives, etc are still possible but have to be done blind. As you can imagine this is frustrating and there is great potential for a mix-up with regard to matching specimens to negative numbers.
I have some questions for you...
Are these CRT data display units unique for this Hitachi model or are units from other models compatible with the H-600? Can CRT units from other electrical equipment be sourced, eg radiology machines? Can new CRT units be bought today that would be compatible? Does anyone have an old working unit that they would be willing to part with? Has anyone had similar experiences with CRT units? Does our unit sound fixable or are we wasting our time?
Thanks,
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia Australia
john.brealey-at-imvs.sa.gov.au
(08) 8222 6612
==============================Original Headers============================== 3, 33 -- From john.brealey-at-imvs.sa.gov.au Wed Oct 11 21:02:16 2006 3, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 3, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C22FMx032484 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 21:02:16 -0500 3, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 3, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9C22FJZ024600 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:32:15 +0930 (CST)' 3, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 3, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9C22FmJ024594 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:32:15 +0930 (CST)' 3, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 3, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id F350934F22 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:32:14 +0930 (CST) 3, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 3, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 3, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 3, 33 -- with LMTP id Yz015DgQqpt4 for {Microscopy-at-MSA.Microscopy.Com} ; 3, 33 -- Thu, 12 Oct 2006 11:32:04 +0930 (CST) 3, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 3, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 89C8F34F19 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:31:28 +0930 (CST) 3, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 3, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 3, 33 -- Subject: Hitachi H-600 CRT Problem 3, 33 -- Date: Thu, 12 Oct 2006 11:31:27 +0930 3, 33 -- Message-ID: {000001c6eda2$549789a0$c88a140a-at-iqe36042} 3, 33 -- MIME-Version: 1.0 3, 33 -- Content-Type: text/plain; 3, 33 -- charset="us-ascii" 3, 33 -- Content-Transfer-Encoding: 7bit 3, 33 -- X-Mailer: Microsoft Office Outlook 11 3, 33 -- Thread-Index: AcbtokEn2wZh7yMrRRGmgQ4kZRU8jA== 3, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================End of - Headers==============================
I run several JEOL TEMs with film cameras (200CX, 2000FX, 2010, 4000) with a large contingent of new users every year. We do get the occasional jam due to misloaded film cassettes by one of the new users. Apart from that we rarely experience camera jams and I would estimate that at the height of our film use the number of films between jams was well in the 10,000's. Film use has declined over the past few years as we have some digital cameras. As far as I can see the camera mechanism has not changed since our 100B so there is no reason to have a different failure rate on any instrument you listed.
If you are experiencing problems with camera jams then check that your cassettes and boxes are not damaged. Check that the films are firmly held in all four corners when they are loaded, very occasionally we get undersized film that will not fit properly. If these are OK then there may be a problem with your camera mechanism, possibly damaged when freeing off a jammed cassette. If you know how to remove the mechanism check that the leaf springs on the pushers are OK and that the tracks are not damaged.
Ron
-----Original Message----- X-from: rcsencsits-at-lbl.gov [mailto:rcsencsits-at-lbl.gov] Sent: 11 October 2006 22:15 To: ron.doole-at-materials.ox.ac.uk
Dear JEOL film camera users:
What is your experience with the film camera? How many successful film cassettes do you average between camera jams? Please quote for each TEM model (100CX, 2000FX, 3100, etc).
Thanks, Roseann
Roseann Csencsits, PhD. Donner TEM Facility Manager Lawrence Berkeley Lab 510-486-4548
==============================Original Headers============================== 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} 7, 21 -- Subject: JEOL camera question 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- X-Mailer: Apple Mail (2.752.3) 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 7, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
==============================Original Headers============================== 20, 26 -- From ron.doole-at-materials.ox.ac.uk Thu Oct 12 02:35:25 2006 20, 26 -- Received: from relay5.mail.ox.ac.uk (relay5.mail.ox.ac.uk [163.1.2.163]) 20, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C7ZO0k016145 20, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 02:35:25 -0500 20, 26 -- Received: from smtp1.mail.ox.ac.uk ([129.67.1.207]) 20, 26 -- by relay5.mail.ox.ac.uk with esmtp (Exim 4.62) 20, 26 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 20, 26 -- id 1GXv6C-0000nq-Gx; Thu, 12 Oct 2006 08:35:24 +0100 20, 26 -- Received: from oums-doole.materials.ox.ac.uk ([129.67.85.58] helo=oumsdoole) 20, 26 -- by smtp1.mail.ox.ac.uk with esmtp (Exim 4.62) 20, 26 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 20, 26 -- id 1GXv6B-0004IX-6L; Thu, 12 Oct 2006 08:35:24 +0100 20, 26 -- From: "Ron Doole" {ron.doole-at-materials.ox.ac.uk} 20, 26 -- To: {rcsencsits-at-lbl.gov} 20, 26 -- Cc: "'Microscopy Society of America'" {Microscopy-at-MSA.Microscopy.Com} 20, 26 -- Subject: RE: [Microscopy] JEOL camera question 20, 26 -- Date: Thu, 12 Oct 2006 08:35:22 +0100 20, 26 -- Message-ID: {000101c6edd0$f9c31510$3a554381-at-oumsdoole} 20, 26 -- MIME-Version: 1.0 20, 26 -- Content-Type: text/plain; 20, 26 -- charset="us-ascii" 20, 26 -- Content-Transfer-Encoding: 7bit 20, 26 -- X-Mailer: Microsoft Office Outlook 11 20, 26 -- Thread-Index: AcbtelS7vQ0g8W45S6mwwOiH18xAxwAUmEuw 20, 26 -- In-Reply-To: {200610112115.k9BLF6PR006189-at-ns.microscopy.com} 20, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
It is not really surprising that you see a difference between the 2 protocols. Please keep in mind that without fixation and with mild permeabilization the antibodies probable have access only to a few compartments of the cell. Using other treatments perhaps allows the antibodies to have better access to the antigens. On the contrary, it is possible that fixation has hidden some antigenic sites. Or perhaps a combination of both effects! Also, be careful using glutaraldehyde in fluorescence since it gives background. I would suggest you try directly in immunoEM and don't lose too much time playing around with protocols because it can be that your antibody simply doesn't work in EM. If you get a signal in EM, you can always discuss its pattern and compare it with the pattern obtained in IF.
Good luck
Stephane
----- Original Message ---- X-from: "kychung2-at-wisc.edu" {kychung2-at-wisc.edu} To: nizets2-at-yahoo.com Sent: Thursday, October 12, 2006 3:43:12 AM
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Email: kychung2-at-wisc.edu Name: Ka Young Chung
Organization: University of Wisconsin-Madison
Title-Subject: [Filtered] Fixation problem
Question: I am working with cardiac myocytes. To perform immunofluorescence, I have permeabilized cells with saponin without fixation, and could get nice signal. The signal was blocked when I used blocking peptide (original immunogen for primary antibody). To look at the localization pattern more in detail, I want to do electron microscopy. In this case, I need to fix the cells. To check whether fixation affects antibody interaction, I fixed cells with 4% paraformaldehyde and 0.1% glutaraldehyde, permeabilized cells with saponin, and then performed immunofluorecence. I saw a permeabilization problem with this experiment so that I tried 0.1% triton instead of saponin. Still, I am not getting the same staining pattern that I got from unfixed cells. How can I solve this problem? Can I just try EM?
An important topic. I'd like to step in with a reply to just the point about "Could aqueous solutions of osmium tetroxide be used in post-fixation steps?" Yes. Not only have I known folks who make their OsO4 in distilled water for TEM, I have studied cultured cells in high-resolution SEM using OsO4 in distilled water, not buffer, for post-fixation. Usually with 1% monomeric tannic acid (for plasma membrane preservation), but not always. Works fine. The colligative properties of the plasma membrane seem to be pretty much eliminate by fixation.
pH isn't the only confusing issue here -- osmotic properties are, too. 3% glut is about 300 mOsM itself, more if there are impurities and after adding buffer. Yet, this hypertonicity doesn't seem to cause osmotic problems.
Worse, something I've never seen discussed: what are the osmotic effects of different pHs? Changing the pH will change the flow of ions and water in and out of cells. If a fixative is present as the pH changes, then the changes in the membrane proteins affecting the osmotic properties will be fixed. So potentionally, a correct pH may in fact not be the physiological pH of the living cell, but the pH that maintains the correct osmotic pressure(s) during fixation.
Phil
} Dear all, } } I have been reading this thread with interest in the hope of learning } something new. The theory of buffers as explained by Bill Tivol, doesn't } seem to be useful when applied to electron microscopy. The reason is that we } judge the effects of different buffers on their effect on morphology (as } Mike Reedy subtly puts it in his message), not on the final pH of the } specimens. } } Judged on buffering capacity alone, sodium cacodylate would fail miserably } as a buffer. The Good buffers (PIPES, HEPES etc) appear to be excellent } buffers for keeping solutions at a stable pH. However, even if PIPES or } HEPES is used to buffer aldehyde, when the solution encounters biological } material there is a sudden drop in pH that is not affected at all by the } buffer in use. Anyone who has fixed cell cultures in the presence of medium } containing pH indicator can attest to the rapid color change that occurs. } } It is worth noting that the pH to which buffers are adjusted, and the pH at } which they are most effective, is not always 7.2 or 7.4. For example, the } use of PIPES at a pH higher than 6.8 will greatly reduce its buffering } capacity. } } I liked the idea of comparing the sizes of fixed cells with living cells to } determine which concentration of buffer is "best" to use. There was no } mention of what the intracellular morphology looked like and if it was } acceptable. I am sure that the best concentration will differ between cell } types. The great disadvantage of this method for determining "correct' } buffer concentrations is that most of our work involves tissues composed of } many different cell types that are all attached to each other. } } Another issue touched on by Mike Reedy is the state of aldehyde-fixed } biological material. Is is really osmotically inactive as assumed (e.g. } "...buffer choice need not be physiological.")? Could aqueous solutions of } osmium tetroxide be used in post-fixation steps? Are aldehyde-fixed } membranes osmotically inactive? } } Our final evaluation of buffers will depend on what we are looking at, or } looking for, in our specimens. Sometimes we need to extract lots of } cytoplasmic components to give high-contrast images. Other times we need to } retain as many proteins as possible so that we can perform } immunocytochemical labeling. Other important considerations for } immunolabeling are the need to retain all antigens at their "natural" site, } yet enable complete antibody accessibility to antigens. } } For the record, our routine fixation protocols involve glutaraldehyde and } sodium cacodylate for morphology, and double-strength aldehyde (formaldehyde } or glutaraldehyde, or mixtures of both) in double strength HEPES buffer for } immunocytochemistry. The double-strength fixative is added to culture } medium, ringer or other physiological medium. There routine fixatives work } well for us, but we do not work with sharks, molluscs or plants. } } Any EM laboratory involved in many projects may find that one fixation } protocol is insufficient for all users. Instead, different protocols } involving different buffers gradually evolve in the lab to cope with each } specimen as needed. What we need is a good understanding of how buffers and } cross-linking chemicals affect the specimens we apply them to. } } 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 } } } } } } } ____________________________________________________________________ } } The ability of a buffer to maintain pH depends on how many groups } there are on the molecule that can take up or release H+ and the pK of } each of these groups. For most buffers there are one or a few of these } groups that have different pKs; e.g. PO4--- with pKs of ~12, ~7, and } ~3. I don't know offhand what pKs are available for either MOPS or } PIPES, but the closer a pK is to the pH you want, the better the buffer } will be at maintaining that pH. The reason for this is easily seen } from the mass action law, which can be written } } K = [A][H]/[AH] } } from which it can be seen that for [H] = K, [A] = [AH], and if [H] = K, } then pH = pK. When the protonated and unprotonated species are at } equal concentration, the addition of a small amount of acid or base } will change the [A]/[AH] ratio less than if they are at unequal } concentration. Therefore, the performance of either Good buffer will } equal that of the other one if the desired pH is the same amount away } from either pK. Another reason to choose one over another is if one of } them affects the process you are trying to perform. It may be that one } will react with your fixative--which I do not think is the case for } MOPS or PIPES and either glut or OsO4. } Yours, } Bill Tivol, PhD } } ___________________________________________________________________ } } For 40 years we have used the physiological or experimental buffer } solution, often with MOPS as the pH buffer, as our main vehicle for } primary fixatives composed with aldehydes, aldehyde-tannic-acid, or } tannic acid alone, because we believe this offers the best hope for } preserving native structure of various muscle structural states (but } also effective with other cells and tissues). "Physiological" mostly } implies proper ions: for extracellular salines (Na+ at ~100-145 mM; } K+ absent or below 2 mM; Mg++ and Ca++ at 1-4 mM); or intracellular } rest-state salines (EGTA and no inadvertent Ca++; Mg at ~5-7 mM; K+ } (or Na+) up to 150 mM if desired; chloride, propionate, acetate, or } methane-sulfonate as anion). } } So for intact frog leg muscle (or insect flight muscle!) we used frog } Ringer solution, for mammalian muscle we used mammalian Ringer, with } either phosphate or MOPS buffer, adjusting pH with a "dirty" } (fixative only) pH electrode to discover the amount of alkali or acid } needed to restore original pH after adding various fixative at } various concentrations. The presence of up to 3 mM calcium and up to } 10 mm magnesium, as dictated by various buffers, was essential in } Ringer recipes, and presented no problem. We were happy because the } fiber x-ray diffraction pattern and the EM itself showed these } fixatives to give good preservation of various distinct pre-fixation } native structural states, and i think this was not so for our brief } long-ago trials of phosphate or s-collidine buffer. We usually } ignored adjustments to osmolarity, but if tried, was always done by } adding the osmolytge to the formulation, sucrose top Ringer, 500,000 } MW dextran to intracellular buffer. } } For secondary fixation, buffer choice need not be physiological. For } aldehyde-only or aldehyde-TA primary fixation, post-fixing in OsO4 } has routinely employed a Maupin-Pollard good-for-actin buffer (0.1M } phosphate, pH 6, with 10 mM MgCl2, used ice cold). } } Most of our work has been with chemically demembranated ("skinned"; } by detergent-glycerol) muscle of rabbit, frog or insect, so we chose } various composition of intracellular buffer to keep the organelles } happy and in the desired functional state of the muscle machinery } (rigor, Mg-ATP relaxed, Mg-ATP-Ca activating, or modified with } nucleotide analogs (like AMPPNP) phosphate analogs (like vanadate). } We have for 30 years used nothing but 20 mM MOPS buffer (relatively } inexpensive, and good for pH 6.5-7.5; we prefer 6.8 for insect) for } both the physiological and fixative versions of our solutions, } typically with 5 mM each of additives like MgCl2, EGTA, Ca-EGTA, ATP, } Na azide (stops microbial growth, arrest ATPase consumption by } mitochondria). For years we strictly used potassium salts in } preference to sodium to honor the cell's intracellular habit, but } lately have slipped away from this dogma without bad results, } drifting to more frequent use of sodium (as hydroxide to adjust pH of } EGTA, ATP, MOPS, etc. Sometimes we use salt to approximate } intracellular ionic strength, typically using KCl (now NaCl) at 100 } or 150 ml, but we often use no extra salt at all, because its } presence or absence has so far made little or no difference to the } fiber x-ray diffraction patterns which are our gold standard for } native and preserved structure. } } We have always steered clear of Tris because glutaraldehyde reacts } with it, changing pH (?? and what else?), and because Tris-buffered } pH alters with changing temperature. } } 0.2% tannic acid alone in the physiological (intracellular type) } buffer works well on skinned cells to fix all but soluble components. } It is blocked or complexed so it becomes unavailable for fixation } when Triton X-100 or PVP are included in the same solution. TA-fix } should be followed after rinse-out by uranyl acetate (typically in DI } water) or OsO4 (in Maupin-Pollard good-for-actin buffer) as a } secondary fixative. } } -mike reedy- } } } ==============================Original Headers============================== } 31, 18 -- From PWebster-at-hei.org Wed Oct 11 16:18:24 2006 } 31, 18 -- Received: from hi0sml1.hei.org } (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) } 31, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k9BLIOUn006955 } 31, 18 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 } 16:18:24 -0500 } 31, 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 ; } 31, 18 -- Wed, 11 Oct 2006 21:18:23 +0000 } 31, 18 -- User-Agent: Microsoft-Entourage/11.2.5.060620 } 31, 18 -- Date: Wed, 11 Oct 2006 14:18:23 -0700 } 31, 18 -- Subject: Re: debating about buffers for TEM fixation } 31, 18 -- From: "Webster, Paul" {PWebster-at-hei.org} } 31, 18 -- To: {microscopy-at-msa.microscopy.com} } 31, 18 -- Message-ID: {C152ADAF.C5ED%PWebster-at-hei.org} } 31, 18 -- Thread-Topic: debating about buffers for TEM fixation } 31, 18 -- Thread-Index: AcbteshdByXfOlluEdua3wANk7Zh7g== } 31, 18 -- Mime-version: 1.0 } 31, 18 -- Content-type: text/plain; } 31, 18 -- charset="US-ASCII" } 31, 18 -- Content-transfer-encoding: 7bit } ==============================End of - Headers==============================
-- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 6, 22 -- From oshel1pe-at-cmich.edu Thu Oct 12 07:19:22 2006 6, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CCJM6W009892 6, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 07:19:22 -0500 6, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 6, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k9CCoWNA010683 6, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:50:41 -0400 6, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 6, 22 -- Thu, 12 Oct 2006 08:19:12 -0400 6, 22 -- Mime-Version: 1.0 6, 22 -- Message-Id: {f06230900c153e0a29915-at-[141.209.160.249]} 6, 22 -- In-Reply-To: {200610112122.k9BLM7xQ014251-at-ns.microscopy.com} 6, 22 -- References: {200610112122.k9BLM7xQ014251-at-ns.microscopy.com} 6, 22 -- Date: Thu, 12 Oct 2006 08:19:10 -0400 6, 22 -- To: Microscopy-at-microscopy.com 6, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 6, 22 -- Subject: [Microscopy] Re: debating about buffers for TEM fixation 6, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 22 -- X-OriginalArrivalTime: 12 Oct 2006 12:19:12.0354 (UTC) FILETIME=[A04A6420:01C6EDF8] 6, 22 -- X-CanItPRO-Stream: default 6, 22 -- X-Spam-Score: -3.9 () L_EXCH_MF,TW_YT 6, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
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Email: U.J.Potter Name: Ursula Potter
Organization: University of Bath
Title-Subject: [Filtered] Neg carriers jamming in JEOL EM
Question: Dear All,
I managed to erase Rosannes message about negative cassettes jamming in JEOL TEM film cameras before I had replied - so hope she sees it here.
We have had this problem and found it was due to warped cassettes. We scratched a very fine number on each cassette and then found it was the same cassette or couple of cassettes causing the problem every time. Since numbering the cassettes and weeding out the troublesome ones we have had very few jamming problems.
Hope this helps.
Ursula ------------------------- CEOS University of Bath UK
I cannot recall having any camera jams with our JEOL 2011. Our older Philips EM420 has periodic jams- usually due to bent film plates or poorly loaded film.
Cheers Roger A. Ristau, PhD Electron Microscopy Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269
} } Dear JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548
==============================Original Headers============================== 6, 20 -- From raristau-at-ims.uconn.edu Thu Oct 12 08:21:16 2006 6, 20 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CDLGk3032294 6, 20 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:21:16 -0500 6, 20 -- Received: from [137.99.20.118] (d20h118.public.uconn.edu [137.99.20.118]) 6, 20 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k9CDL9PQ019634 6, 20 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 09:21:09 -0400 6, 20 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 20 -- Date: Thu, 12 Oct 2006 09:20:03 -0400 6, 20 -- Subject: Re: [Microscopy] JEOL camera question 6, 20 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 6, 20 -- To: {microscopy-at-microscopy.com} 6, 20 -- Message-ID: {C153B943.17B7%raristau-at-ims.uconn.edu} 6, 20 -- In-Reply-To: {200610112114.k9BLEOoS004677-at-ns.microscopy.com} 6, 20 -- Mime-version: 1.0 6, 20 -- Content-type: text/plain; charset="US-ASCII" 6, 20 -- Content-transfer-encoding: 7bit 6, 20 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 6, 20 -- X-UConn-MailScanner: Found to be clean 6, 20 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
Roseann I have a JEOL 100 CX II, that was installed in 1982. I've been its guardian since 1988. The only time I experience camera jams is when one of my users decides to be "helpful" and reload the cassettes for me. Inevitably, one plate in the pile will have a loose corner that catches in the mechanism. As a rule, I just don't let anyone other than me and my technician load the film, and we are fine. A little care in the darkroom goes a long way. 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, 23 -- From lcgould-at-med.cornell.edu Thu Oct 12 08:23:12 2006 1, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CDNBwr003031 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:23:11 -0500 1, 23 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9CDN0gA013861 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 09:23:10 -0400 (EDT) 1, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 23 -- by mpx1.med.cornell.edu 1, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 23 -- with ESMTPA id {0J7000HS4YI1G070-at-mpx1.med.cornell.edu} for 1, 23 -- microscopy-at-microscopy.com; Thu, 12 Oct 2006 09:23:00 -0400 (EDT) 1, 23 -- Date: Thu, 12 Oct 2006 09:22:47 -0400 1, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 23 -- Subject: Re: [Microscopy] JEOL camera question 1, 23 -- In-reply-to: {200610112107.k9BL7bSs029708-at-ns.microscopy.com} 1, 23 -- Sender: lcgould-at-med.cornell.edu 1, 23 -- To: rcsencsits-at-lbl.gov, Microscopy Listserver {microscopy-at-microscopy.com} 1, 23 -- Message-id: {p06230902c153f112eaef-at-[140.251.48.23]} 1, 23 -- MIME-version: 1.0 1, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 23 -- References: {200610112107.k9BL7bSs029708-at-ns.microscopy.com} 1, 23 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.12.54942 ==============================End of - Headers==============================
Hi Roseann, I also have an ancient 120CX (installed 1979) as well as a newer 2011. I did have problems with the camera on the old machine occasionally (perhaps once every 10,000 plates or so), and the problem was invariably caused by my putting the film carriers in the wrong way round or not making sure the film was in the carrier properly. I find concentration does tend to drift after the first few thousand... We ran two film boxes with 50 plates, but just filling the empty one with the plates we had just taken, typically between 10 and 20 at a time. So I guess we had a problem maybe once every 500 cassette swaps. However getting the thing unstuck did also cause damage to one of the 'nails' once, and I took the camera unit out of the microscope to fix it. (Also, I have to admit, I was curious as to how it worked!) If you are having a lot of problems it may be worth it for you - it is not difficult and can be done with the machine running. It has been a while but I can give you some instructions if you want. When you have the unit out of the microscope you can feed through all of your carriers one at a time, if you like, and see if any particular ones are causing a problem. I have no problems with the 2011 camera, but I have taken less than 20 negatives in 3 years (all digital)!
Cheers
Richard
} } Dear JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Thu Oct 12 08:52:47 2006 8, 31 -- Received: from mail115.messagelabs.com (mail115.messagelabs.com [195.245.231.179]) 8, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9CDqjFa021518 8, 31 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:52:46 -0500 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-5.tower-115.messagelabs.com!1160660918!12701429!17 8, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 29680 invoked from network); 12 Oct 2006 13:51:00 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-5.tower-115.messagelabs.com with SMTP; 12 Oct 2006 13:51:00 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 8, 31 -- Thu, 12 Oct 2006 14:54:06 +0100 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: [Microscopy] Re: JEOL camera question 8, 31 -- Date: Thu, 12 Oct 2006 14:54:09 +0100 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E24316A-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: [Microscopy] Re: JEOL camera question 8, 31 -- Thread-Index: AcbuBePVT7HyYiPWTHe1COvrq2B6iw== 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 12 Oct 2006 13:54:06.0549 (UTC) FILETIME=[E24B9850:01C6EE05] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CDqjFa021518 ==============================End of - Headers==============================
I have been trying to help a student do Cryo-SEM on cultured T-cells - he extracts human blood, collects T-cells, cultures them in a medium containing serum and then exposes them for varying lengths of time to chemokines. After culturing the cells were washed briefly in PBS (to remove serum)then fixed in 1% glutaraldehyde in PBS for 15mins then applied to a filter where as much liquid as possible is removed immediately before freezing in nitrogen slush and carrying out Cryo-SEM. We have the problem that the cells appear in the SEM round, smooth & without psuedopodia!
We have previously done TEM on the same cells (different extraction) and had good results showing cells with interesting psuedopodia. The difference with the TEM prep was that the fixative was added to the culture medium and the cells fixed for 2hrs at room temp then overnight in the fridge.
We intend to try conventional SEM on the cells next using the initial TEM prep method then Critical Point Drying after dehydration. However I am concerned that serum will be fixed to the surface of the cells if they are not washed prior to fixation.
I would appreciate any advice on the above.
Regards Ursula ------------------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
==============================Original Headers============================== 8, 23 -- From U.J.Potter-at-bath.ac.uk Thu Oct 12 09:57:06 2006 8, 23 -- Received: from kelly.bath.ac.uk (kelly.bath.ac.uk [138.38.32.20]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CEv5hY004739 8, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 09:57:05 -0500 8, 23 -- Received: from mary.bath.ac.uk ([138.38.32.14] ident=mmdf) 8, 23 -- by kelly.bath.ac.uk with smtp id 1GY1zc-0006UW-4i 8, 23 -- for Microscopy-at-microscopy.com 8, 23 -- (return-path {U.J.Potter-at-bath.ac.uk} ); Thu, 12 Oct 2006 15:57:04 +0100 8, 23 -- Received: from eapc-03.campus.bath.ac.uk 8, 23 -- ( eapc-03.campus.bath.ac.uk [138.38.136.63] ) by bath.ac.uk 8, 23 -- id aa17613 for {Microscopy-at-microscopy.com} ; 12 Oct 2006 15:57 +0100 8, 23 -- Date: Thu, 12 Oct 2006 15:57:02 +0100 8, 23 -- From: Ursula Potter {U.J.Potter-at-bath.ac.uk} 8, 23 -- To: Microscopy-at-microscopy.com 8, 23 -- Subject: SEM of lymphocytes 8, 23 -- Message-ID: {24575875.1160668622-at-eapc-03.campus.bath.ac.uk} 8, 23 -- Originator-Info: login-id=mssujp; server=imaphost.bath.ac.uk 8, 23 -- X-Mailer: Mulberry/3.1.0 (Win32) 8, 23 -- MIME-Version: 1.0 8, 23 -- Content-Type: text/plain; charset=us-ascii; format=flowed 8, 23 -- Content-Transfer-Encoding: 7bit 8, 23 -- Content-Disposition: inline 8, 23 -- X-Scanner: f2dba285a45b48b55d24015519db9286d4c05056 ==============================End of - Headers==============================
I have learnt a lot about buffers used in fixative and in general about buffers in sample preparation. This is very helpful discussion. We work with mollusks (Oysters) and our oysters are reared at 860 mOSM of sea water, and a pH of near about 8.2, the cells I look at in SEM/TEM have very fine pseudopodia and the preservation of this structure in my primary concern. Could any one please suggest a buffer that works best at a pH higher than 8? After making up the buffers we measure the osmolality and adjust it, but now from the discussion I feel the need to know a buffer that can work at a higher pH.
Thank you,
Neeraj V. Gohad.
Graduate Research Assistant Department of Biological Sciences, 132 Long Hall, Clemson University, Clemson, SC-29634
==============================Original Headers============================== 7, 21 -- From ngohad-at-CLEMSON.EDU Thu Oct 12 10:43:01 2006 7, 21 -- Received: from CLEMSON.EDU (mail.clemson.edu [130.127.28.87]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CFh08I017917 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Thu, 12 Oct 2006 10:43:01 -0500 7, 21 -- Received: from X ([130.127.120.198]) 7, 21 -- by CLEMSON.EDU (8.13.6/8.13.1) with ESMTP id k9CFguVl005242 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Thu, 12 Oct 2006 11:42:56 -0400 (EDT) 7, 21 -- Message-Id: {200610121542.k9CFguVl005242-at-CLEMSON.EDU} 7, 21 -- From: "Neeraj Gohad" {ngohad-at-CLEMSON.EDU} 7, 21 -- To: {Microscopy-at-Microscopy.Com} 7, 21 -- Subject: SEM/TEM-Buffer for a higher pH 7, 21 -- Date: Thu, 12 Oct 2006 11:42:53 -0400 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 7, 21 -- Thread-Index: AcbuFRR3p35jtat0TL27mVpV2pBoxw== 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 7, 21 -- X-Spam-Level: 7, 21 -- X-Scanned-By: mail mimedefang 2.56 on 130.127.28.87 ==============================End of - Headers==============================
Neeraj, In the late 1970's I worked with Mitilus and Thyone sperm in the lab of Lew Tilney. At that time the primary fixative that I used was 1% glutaraldehyde, if I remember correctly, in filtered sea water. The samples were then washed in Millonig's Phosphate Buffer and post fixed in 1% OsO4 in Phosphate Buffer -at- a pH 6.2. on ice and in the dark.
My initial reaction when I received the protocol was very strong but it worked well in this system. It may work for you also.
Pat Connelly EM Core Facility NHLBI/NIH Bethesda, MD 20892 301-496-3491
-----Original Message----- X-from: ngohad-at-CLEMSON.EDU [mailto:ngohad-at-CLEMSON.EDU] Sent: Thursday, October 12, 2006 11:52 AM To: Connelly, Patricia (NIH/NHLBI) [E]
Roseann, I just had a jam on a 100CX II caused by someone loading a plate in backwards and subsequent damage to a spring. This was the first in at least a year with 40--50 film loads and a couple thousand films exposed. I would rate the system as very reliable. Larry
rcsencsits-at-lbl.gov 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 JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548 } } } } } ==============================Original Headers============================== } 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 } 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) } 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 } 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) } 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; } 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) } 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) } 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} } 7, 21 -- Content-Transfer-Encoding: 7bit } 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} } 7, 21 -- Subject: JEOL camera question } 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 } 7, 21 -- To: Microscopy-at-Microscopy.Com } 7, 21 -- X-Mailer: Apple Mail (2.752.3) } 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 } 7, 21 -- X-Virus-Status: Clean } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 5, 30 -- From Larry.Ackerman-at-ucsf.edu Thu Oct 12 12:15:28 2006 5, 30 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CHFQ3A008651 5, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 12:15:27 -0500 5, 30 -- Received: from 64.54.128.152 by emfmcb02.ucsfmedicalcenter.org with 5, 30 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 5, 30 -- Thu, 12 Oct 2006 10:27:03 -0700 5, 30 -- X-Server-Uuid: 44B63953-633F-4500-B2DA-A3E478718104 5, 30 -- Received: from [128.218.123.88] ([128.218.123.88]) by 5, 30 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 Oct 5, 30 -- 2006 10:15:19 -0700 5, 30 -- Message-ID: {452E7820.6010100-at-ucsf.edu} 5, 30 -- Date: Thu, 12 Oct 2006 10:15:12 -0700 5, 30 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 5, 30 -- Reply-to: larry.ackerman-at-ucsf.edu 5, 30 -- Organization: UCSF, NeuroAnatomy 5, 30 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 5, 30 -- X-Accept-Language: en-us, en 5, 30 -- MIME-Version: 1.0 5, 30 -- To: Microscopy-at-microscopy.com 5, 30 -- Subject: Re: [Microscopy] JEOL camera question 5, 30 -- References: {200610112111.k9BLBTYB004083-at-ns.microscopy.com} 5, 30 -- In-Reply-To: {200610112111.k9BLBTYB004083-at-ns.microscopy.com} 5, 30 -- X-OriginalArrivalTime: 12 Oct 2006 17:15:19.0319 (UTC) 5, 30 -- FILETIME=[FE3A1A70:01C6EE21] 5, 30 -- X-WSS-ID: 6930A56D1IO192507-01-01 5, 30 -- Content-Type: text/plain; 5, 30 -- charset=iso-8859-1; 5, 30 -- format=flowed 5, 30 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
On Oct 12, 2006, at 8:43 AM, ngohad-at-CLEMSON.EDU wrote:
} I have learnt a lot about buffers used in fixative and in general about } buffers in sample preparation. This is very helpful discussion. We } work } with mollusks (Oysters) and our oysters are reared at 860 mOSM of sea } water, } and a pH of near about 8.2, the cells I look at in SEM/TEM have very } fine } pseudopodia and the preservation of this structure in my primary } concern. } Could any one please suggest a buffer that works best at a pH higher } than 8? } After making up the buffers we measure the osmolality and adjust it, } but now } from the discussion I feel the need to know a buffer that can work at a } higher pH. } Dear Neeraj, I don't have access to a table of the pKs of all the Good buffers, but I seem to remember that there are some with pKs in the range you want. Failing that, the Handbook of Chemistry and Physics gives the pH for a .1N solution of NaCO3 as 8.4--of course, many cations will precipitate bicarbonate--morpholine has a pK of 8.33, several dipeptides have pKs in this range with leucylglycine at 8.28, and many alkaloids have pKs near 8.2, but explaining why you want morphine (8.21), strychnine (8.26), or codeine (8.21) may not be worth it. 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 Oct 12 12:32:28 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 (8.12.11.20060308/8.12.8) with ESMTP id k9CHWSdd019399 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 12:32:28 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id A3DD310A160 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 10:32:27 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id C2F60109DCC 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 10:32:25 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200610121543.k9CFhGta018085-at-ns.microscopy.com} 4, 22 -- References: {200610121543.k9CFhGta018085-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {08661d9a0eb4733694944758640e4291-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] SEM/TEM-Buffer for a higher pH 4, 22 -- Date: Thu, 12 Oct 2006 10:36:32 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Try skipping the glut fix. You're cryofixing the cells, so a chemical fixation should not also be needed. I assume the cells are cultured on something nice and thin and heat conductive, yes? Little metal coupons, formvar-coated TEM grids, bits of coverslip sized to the SEM stage before culturing, or the like. Rinse the cells briefly with serum-free buffer, best is the same buffer they're cultured in, just without serum. Make sure the temperature of the wash buffer is the same as the temperature of the incubation medium. Remove from the buffer and immediatey plunge into the slush nitrogen. Don't blot off the excess buffer too much -- just quick touch of the edge of the grid, etc.. There won't be much, and rapid plunging into slush nitrogen freezes fast enough that a thin layer of buffer won't interfer with good freezing. The buffer water gets vacuum sublimated during cryocoating, and if you're looking at the cells uncoated, they can still be sublimated. The conventional fixation/CPD method also works well, but I strongly suggest adding 1% monomeric tannic acid to the 1% glutaraldehyde as well as to the osmium (although OsO4 is not necessarily needed for high-resolution low-voltage SEM of cells). You are correct, the serum must be washed away however you fix the cells. Otherwise, you get these wonderful, stringy strands of serum proteins obscuring everything. Especially with chemical fixation.
Phil
} Dear All, } } I have been trying to help a student do Cryo-SEM on cultured T-cells - he } extracts human blood, collects T-cells, cultures them in a medium } containing serum and then exposes them for varying lengths of time to } chemokines. After culturing the cells were washed briefly in PBS (to } remove serum)then fixed in 1% glutaraldehyde in PBS for 15mins then applied } to a filter where as much liquid as possible is removed immediately before } freezing in nitrogen slush and carrying out Cryo-SEM. We have the problem } that the cells appear in the SEM round, smooth & without psuedopodia! } } We have previously done TEM on the same cells (different extraction) and } had good results showing cells with interesting psuedopodia. The difference } with the TEM prep was that the fixative was added to the culture medium and } the cells fixed for 2hrs at room temp then overnight in the fridge. } } We intend to try conventional SEM on the cells next using the initial TEM } prep method then Critical Point Drying after dehydration. However I am } concerned that serum will be fixed to the surface of the cells if they are } not washed prior to fixation. } } I would appreciate any advice on the above. } } Regards } Ursula } ------------------------ } } } Ursula J. Potter } Centre for Electron Optical Studies (CEOS) } Building 3 West 2.15 } The University of Bath } Claverton Down } Bath BA2 7AY } UK } Tel: 01225 385651 } Email: U.J.Potter-at-bath.ac.uk -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Thu Oct 12 12:49:15 2006 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CHnFVO030148 4, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 12:49:15 -0500 4, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k9CIKRNC014291 4, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:20:35 -0400 4, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 22 -- Thu, 12 Oct 2006 13:49:09 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f0623090ac1542d4c90e1-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200610121502.k9CF2P6F012736-at-ns.microscopy.com} 4, 22 -- References: {200610121502.k9CF2P6F012736-at-ns.microscopy.com} 4, 22 -- Date: Thu, 12 Oct 2006 13:49:07 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] SEM of lymphocytes 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 12 Oct 2006 17:49:09.0764 (UTC) FILETIME=[B8776C40:01C6EE26] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -3.4 () J_CHICKENPOX_54,L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Dear All, I got a mimosa as a present and I am very fascinated by this plant. I would like to study the function of the movement of the leaves with SEM and TEM. Is anybody out there who has a fixation protocoll or who did work on this plant? Any images available in the net?
Terrific little article on buffers, Good (= Good's) buffers with table or working range and pKs in wikipediat at http://stanxterm.aecom.yu.edu/wiki/index.php?page=About_buffers -mike-
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************************ 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
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
We are about to scrap an Akashi 002A TEM and a 1970's vintage Balzers BAF300 freeze fracture apparatus. Before we do however, is there anyone out there who would like any specific parts or components from either of these instruments. Perhaps to keep one you have going.
You pay the cost of freight.
Regards
Allan
Allan Mitchell Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
==============================Original Headers============================== 8, 20 -- From allan.mitchell-at-stonebow.otago.ac.nz Thu Oct 12 14:54:58 2006 8, 20 -- Received: from mailhub1.otago.ac.nz (mailhub1.otago.ac.nz [139.80.64.218]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJsvuI009906 8, 20 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 14:54:58 -0500 8, 20 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 8, 20 -- by mailhub1.otago.ac.nz (8.13.6/8.13.6) with ESMTP id k9CJsuWS025781 8, 20 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 08:54:56 +1300 8, 20 -- Received: from allan.otago.ac.nz ([139.80.40.92]) 8, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 8, 20 -- id 1GY6ds-0007pR-Bf 8, 20 -- for microscopy-at-msa.microscopy.com; Fri, 13 Oct 2006 08:54:56 +1300 8, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- Message-Id: {CDB10E09-AE2C-475A-8D71-A6D99D5291F6-at-stonebow.otago.ac.nz} 8, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 20 -- To: microscopy-at-msa.microscopy.com 8, 20 -- From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} 8, 20 -- Subject: Equipment being disposed of. 8, 20 -- Date: Fri, 13 Oct 2006 08:54:55 +1300 8, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Our facility is used both for research and pathology/hospital work. We have a digital camera with telescope (!) software for image collection which I find truly frustrating. You should get the highest resolution camera that you can (the pathologists here want 2K by 2K), but in my opinion you should also give the software interface high priority. I used an AMT camera at my last position and was very happy with the simplicity of use as well as the (virtually) live imaging capabilities. Most people hardly used the fluorescent screen once set up with the camera. Taking a picture was a matter of pushing an onscreen "button" and what you saw is what you got. The software was also calibrated with the microscope and didn't require manually annotating each image with the scale bar. My big complaint with that system was it was only 1K by 1K but you can easily do much better now. Kim
GBurgess-at-exchange.hsc.mb.ca 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 TEM users, } We are a pathology/hospital based TEM facility equipped with a Jeol 1010 } electron microscope. We would like to change to a digital camera system, } since we find that wet-film technology is slow and expensive. I am } hoping that there are users out there that can provide me with opinions } about digital camera systems to help educate me about what they like, } and what they think is important in a TEM digital camera and its } associated computer [hardware/software] system. } } I have never used a TEM digital camera, so I have no experience as such } with any digital camera system, other than consumer digital cameras, so } it's hard for me to judge the merits of a digital camera without } "getting my feet wet" so to speak, and trying it out. } } I'm open to suggestions, thoughts, and opinions of experienced users or } any comments on this topic. } } } Garry Burgess } } Charge Technologist } Electron Microscopy } Department of Pathology } Winnipeg, Canada } }
-- Kim Rensing Ph.D. Manager, Microscopy and Imaging Facility, University of Calgary, Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1 O: 403-220-3488 F: 403-270-8928
==============================Original Headers============================== 4, 28 -- From krensing-at-ucalgary.ca Thu Oct 12 15:30:32 2006 4, 28 -- Received: from mr2.ucalgary.ca (mr2.ucalgary.ca [136.159.34.166]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CKUVDg021084 4, 28 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 15:30:32 -0500 4, 28 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 4, 28 -- by mr2.ucalgary.ca (Postfix) with ESMTP id 5A8413667D; 4, 28 -- Thu, 12 Oct 2006 14:30:31 -0600 (MDT) 4, 28 -- Received: from [192.168.0.100] ([136.159.164.171]) 4, 28 -- (authenticated (0 bits)) 4, 28 -- by smtp2.ucalgary.ca (8.11.7/8.11.6) with ESMTP id k9CKUR028214 4, 28 -- (using TLSv1/SSLv3 with cipher DHE-RSA-AES256-SHA (256 bits) verified NO); 4, 28 -- Thu, 12 Oct 2006 14:30:27 -0600 4, 28 -- Message-ID: {452EA5E0.9020107-at-ucalgary.ca} 4, 28 -- Date: Thu, 12 Oct 2006 14:30:24 -0600 4, 28 -- From: Kim Rensing {krensing-at-ucalgary.ca} 4, 28 -- Organization: Microscopy and Imaging, U of C 4, 28 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 4, 28 -- MIME-Version: 1.0 4, 28 -- To: GBurgess-at-exchange.hsc.mb.ca, microscopy-at-microscopy.com 4, 28 -- Subject: Re: [Microscopy] TEM Digital Camera - Opinions of Important Features 4, 28 -- Sought 4, 28 -- References: {200610121954.k9CJs3qs009368-at-ns.microscopy.com} 4, 28 -- In-Reply-To: {200610121954.k9CJs3qs009368-at-ns.microscopy.com} 4, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 28 -- Content-Transfer-Encoding: 7bit 4, 28 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 4, 28 -- X-UCalgary-MailScanner: Found to be clean 4, 28 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
I'm sure you are going to get a number of answers to your question. As a manufacturer of these cameras, let me point out a few things that I think might help you. I don't want to make this into a "commercial" so I will stay away from specifics and try to provide some generic information that applies to all manufacturers. If some of this sounds too commercial, I apologize. It is not my intention.
1) TEM cameras come in two flavors: Bottom mounted and Side-mounted. The bottom-mount cameras are in general better for high-resolution imaging, while the side-mount cameras give you a better field of view. However, I would not automatically dismiss the bottom-mount cameras, as there are ways to overcome the smaller field of view (automatic image montaging), and you might find that a bottom-mount camera is better for your needs. Bottom-mount cameras are typically more expensive, so your budget will play a significant role.
2) Throughput. Most pathology departments we have talked to put a premium on efficiency so that they can work on as many cases as possible per day. If that is the case for you also, the live speed of the camera is important. It is a lot faster if you can work with the camera in a live mode, then simply push a button to snap a picture, than to work on the microscope in the traditional way (binoculars), then switch to the camera (insert in beam or lift screen), then wait for a few seconds to take a picture. If the picture didn't work out, you would have to go back to the binoculars, adjust the microscope, and start over again. A reasonable fast camera (10 frames per second) let you work on the screen computer screen directly.
3) Organization. Since digital images come cheap, users tend to take more pictures. If you have multiple users it can get quite busy on the hard disk, and you will have to implement some structure to keep users, cases, tissues, etc. apart. Here it might be advantageous to have a way of organizing your images in a database. If that is important to you, make sure that you are not missing out on that later.
4) Other instruments: Most labs have other instruments that are used for the same sample. For example, a grossing station for taking pictures of the tissue and a light microscope for color pictures. If you have that, it might be advantageous to select a system that allows information sharing and the same user interface between electron microscope and other instruments.
5) Hospital. Recently, there has been a push by pathologists to use workflow oriented software for their work, similar to what the radiologists have done for years. If this applies to you, make sure that you have an option to interface with HIS or PACS systems.
6) Reports: In most pathology labs, a technician acquires the images and hands them to the pathologist who then makes a diagnosis based on the images. This can be a critical step. Some pathologists are very particulate how the images need to look like (contrast, brightness, magnification, even paper), others work with multiple formats. Whatever is the case, you need to anticipate this. If the pathologist insists on paper prints in a certain format, the printer becomes an important part of the system. It needs to be fast, and it has to have the right tint. That's not easy to find and you need to talk to the system manufacturers for help. You might also ask the vendors about report capabilities of their software. Predefined and customized reports make it a lot easier to have the "right" format for the pathologist. A digital system, however, allows to get rid of the paper altogether. If you use a database for your images, you can probably install some software on the pathologists desktop computer, they can quickly find the images and make a diagnosis on the computer. If you think that this might be a possibility, ask the vendors about this option.
7) Support: Make sure that you can get good support for your system. Nothing is more frustrating than having to go back to film because you can't get the support you need.
8) Microscope tuning: For newer instruments, there is often an option for tuning the microscope automatically. This can help save time and make imaging easier for new staff. If that is important to you, find out if the software supports tuning the microscope.
9) Advanced features: If you require advanced features such as Tomography, make sure the system you buy supports it, or at least supports a file format that can be accepted by the third party software you use for the advanced features. In most pathology situations this is not of prime importance, though.
Ok, that's all I want to say right now. Sorry for rambling...
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: GBurgess-at-exchange.hsc.mb.ca [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, October 12, 2006 13:53 To: Mike Bode
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
==============================Original Headers============================== 32, 24 -- From Mike.Bode-at-olympus-sis.com Thu Oct 12 15:40:07 2006 32, 24 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 32, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CKe6wK031719 32, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 15:40:06 -0500 32, 24 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 32, 24 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id k9CKl3qZ019917 32, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 22:47:06 +0200 32, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 32, 24 -- Content-class: urn:content-classes:message 32, 24 -- MIME-Version: 1.0 32, 24 -- Content-Type: text/plain; 32, 24 -- charset="us-ascii" 32, 24 -- Subject: RE: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 32, 24 -- Date: Thu, 12 Oct 2006 22:37:29 +0200 32, 24 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC9454DBA6-at-ms-s-gws.soft-imaging.net} 32, 24 -- In-Reply-To: {200610121953.k9CJrRjQ008086-at-ns.microscopy.com} 32, 24 -- X-MS-Has-Attach: 32, 24 -- X-MS-TNEF-Correlator: 32, 24 -- Thread-Topic: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 32, 24 -- Thread-Index: AcbuOBeAXmKp2X+iQ5ulvAiUn8EiPgAASjIA 32, 24 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 32, 24 -- To: {Microscopy-at-microscopy.com} 32, 24 -- Content-Transfer-Encoding: 8bit 32, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CKe6wK031719 ==============================End of - Headers==============================
Hi All, By early next year, we will have a surplus Cambridge 250 Mk3 SEM with a Link model E5431 EDS detector [all purchased in 1986] available very cheap to a good home! This SEM is still in operating condition and may be of interest to anyone who is still using an SEM of this model for spare parts, etc. Contact me if you are interested in this equipment. You would have to pay the cost of freight. Regards,
Doug Hopcroft Electron Microscope Unit, Institute of Molecular BioSciences, Massey University, Private Bag 11 222, Palmerston North, New Zealand. Phone 06 356 9099 ext.81098
==============================Original Headers============================== 4, 17 -- From d.hopcroft-at-massey.ac.nz Thu Oct 12 15:45:14 2006 4, 17 -- Received: from its-mail1.massey.ac.nz (its-mail1.massey.ac.nz [130.123.128.11]) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CKjDVB006529 4, 17 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 15:45:13 -0500 4, 17 -- Received: from TUR-MM3.massey.ac.nz (tur-mm3.massey.ac.nz [130.123.128.140]) 4, 17 -- by its-mail1.massey.ac.nz (8.9.3/8.9.3) with ESMTP id JAA03815 4, 17 -- for {microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 09:45:11 +1300 (NZDT) 4, 17 -- Received: from IT041057.massey.ac.nz (Not Verified[130.123.143.35]) by TUR-MM3.massey.ac.nz with NetIQ MailMarshal 4, 17 -- id {B452ea9570001} ; Fri, 13 Oct 2006 09:45:11 +1300 4, 17 -- Message-Id: {6.2.1.2.0.20061013092853.031abb80-at-pop3.massey.ac.nz} 4, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 4, 17 -- Date: Fri, 13 Oct 2006 09:45:11 +1300 4, 17 -- To: microscopy-at-microscopy.com 4, 17 -- From: Doug Hopcroft {d.hopcroft-at-massey.ac.nz} 4, 17 -- Subject: Surplus Cambridge 250 Mk3 SEM. 4, 17 -- Mime-Version: 1.0 4, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Can someone suggest help with sections falling off of Ni grids during Immuno gold staining? Grids are clean and sections are dried overnight to 3 days prior to staining. I have not yet tried a pap pen to adhere the sections...but I'm looking at my "Coat-Quick "G" grid coating pen and beginning to ask.... Why not? Any reasons not to use it?
Thanks, Linda
Linda M. Fox Loyola University Stritch School of Medicine Core Imaging Facility 2160 S. First Ave. Maywood, Il 60153 Bld. 102 Room 0617 1-708-216-3395 lfox1-at-lumc.edu
==============================Original Headers============================== 7, 17 -- From lfox1-at-lumc.edu Thu Oct 12 16:14:08 2006 7, 17 -- Received: from GWPRIMARY.luhs.org (gw5gate.lumc.edu [147.126.200.222]) 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CLE8IU021017 7, 17 -- for {Microscopy-at-Microscopy.Com} ; Thu, 12 Oct 2006 16:14:08 -0500 7, 17 -- Received: from Primary-MTA by GWPRIMARY.luhs.org 7, 17 -- with Novell_GroupWise; Thu, 12 Oct 2006 16:14:14 -0500 7, 17 -- Message-Id: {s52e69d6.030-at-GWPRIMARY.luhs.org} 7, 17 -- X-Mailer: Novell GroupWise Internet Agent 6.5.4 7, 17 -- Date: Thu, 12 Oct 2006 16:13:36 -0500 7, 17 -- From: "Linda Fox" {lfox1-at-lumc.edu} 7, 17 -- To: {Microscopy-at-Microscopy.Com} 7, 17 -- Subject: ICC gold and pap pens? 7, 17 -- Mime-Version: 1.0 7, 17 -- Content-Type: text/plain; charset=US-ASCII 7, 17 -- Content-Disposition: inline 7, 17 -- Content-Transfer-Encoding: 8bit 7, 17 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CLE8IU021017 ==============================End of - Headers==============================
There's also a useful booklet from Calbiochem called (you guessed it) Buffers: A guide for the preparation and use of buffers in biological systems. I have the "new" 1995 edition, but it has all the Good buffers, recipes, comments about other issues such as cation chelation, notes on protein pI, etc. Rosemary
} From: mike.reedy-at-cellbio.duke.edu } Reply-To: mike.reedy-at-cellbio.duke.edu } Date: Thu, 12 Oct 2006 13:32:18 -0500 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] Re: SEM/TEM-Buffer for a higher pH } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Terrific little article on buffers, Good (= Good's) buffers with } table or working range and pKs in wikipediat at } http://stanxterm.aecom.yu.edu/wiki/index.php?page=About_buffers } -mike- } } } ---------------------------------------------------------------------------- } } 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 } } ---------------------------------------------------------------------------- } } } } } } On Oct 12, 2006, at 8:43 AM, ngohad-at-CLEMSON.EDU wrote: } } } } } I have learnt a lot about buffers used in fixative and in general about } } } buffers in sample preparation. This is very helpful discussion. We } } } work } } } with mollusks (Oysters) and our oysters are reared at 860 mOSM of sea } } } water, } } } and a pH of near about 8.2, the cells I look at in SEM/TEM have very } } } fine } } } pseudopodia and the preservation of this structure in my primary } } } concern. } } } Could any one please suggest a buffer that works best at a pH higher } } } than 8? } } } After making up the buffers we measure the osmolality and adjust it, } } } but now } } } from the discussion I feel the need to know a buffer that can work at a } } } higher pH. } } } } } Dear Neeraj, } } I don't have access to a table of the pKs of all the Good buffers, but } } I seem to remember that there are some with pKs in the range you want. } } Failing that, the Handbook of Chemistry and Physics gives the pH for a } } .1N solution of NaCO3 as 8.4--of course, many cations will precipitate } } bicarbonate--morpholine has a pK of 8.33, several dipeptides have pKs } } in this range with leucylglycine at 8.28, and many alkaloids have pKs } } near 8.2, but explaining why you want morphine (8.21), strychnine } } (8.26), or codeine (8.21) may not be worth it. } } 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 Oct 12 12:32:28 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 (8.12.11.20060308/8.12.8) with } } ESMTP id k9CHWSdd019399 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 } } 12:32:28 -0500 } } 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) } } 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id A3DD310A160 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 } } 10:32:27 -0700 (PDT) } } 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu } } [131.215.2.133]) } } 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id C2F60109DCC } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 } } 10:32:25 -0700 (PDT) } } 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) } } 4, 22 -- In-Reply-To: {200610121543.k9CFhGta018085-at-ns.microscopy.com} } } 4, 22 -- References: {200610121543.k9CFhGta018085-at-ns.microscopy.com} } } 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } } 4, 22 -- Message-Id: {08661d9a0eb4733694944758640e4291-at-caltech.edu} } } 4, 22 -- Content-Transfer-Encoding: 7bit } } 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} } } 4, 22 -- Subject: Re: [Microscopy] SEM/TEM-Buffer for a higher pH } } 4, 22 -- Date: Thu, 12 Oct 2006 10:36:32 -0700 } } 4, 22 -- To: microscopy-at-msa.microscopy.com } } 4, 22 -- X-Mailer: Apple Mail (2.624) } } 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 } } ==============================End of - Headers============================== } } } -- } -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 } } Office 919-668-2534 } Lab 919-684-5674 } Fax 919-681-9929 } mike.reedy-at-cellbio.duke.edu } http://note.cellbio.duke.edu/Faculty/Research/Reedy.html } } ==============================Original Headers============================== } 6, 22 -- From mike.reedy-at-cellbio.duke.edu Thu Oct 12 13:29:29 2006 } 6, 22 -- Received: from porthos.duhs.duke.edu (porthos.duhs.duke.edu } [152.16.199.201]) } 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k9CITTBg019570 } 6, 22 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 13:29:29 -0500 } 6, 22 -- Received: from cellbio.duke.edu (voice.cellbio.duke.edu } [152.16.14.127]) } 6, 22 -- by porthos.duhs.duke.edu (8.13.4/8.13.4) with ESMTP id } k9CIT6E91249430 } 6, 22 -- (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NO); } 6, 22 -- Thu, 12 Oct 2006 14:29:06 -0400 } 6, 22 -- Received: from [152.16.192.164] (HELO [152.16.6.135]) } 6, 22 -- by cellbio.duke.edu (CommuniGate Pro SMTP 4.1.8) } 6, 22 -- with ESMTP id 4263121; Thu, 12 Oct 2006 14:29:05 -0400 } 6, 22 -- Mime-Version: 1.0 } 6, 22 -- Message-Id: {p06210218c15439705d76-at-[152.16.6.135]} } 6, 22 -- In-Reply-To: {200610121736.k9CHarYO025302-at-ns.microscopy.com} } 6, 22 -- References: {200610121736.k9CHarYO025302-at-ns.microscopy.com} } 6, 22 -- Date: Thu, 12 Oct 2006 14:29:00 -0400 } 6, 22 -- To: tivol-at-caltech.edu } 6, 22 -- From: Mike Reedy {mike.reedy-at-cellbio.duke.edu} } 6, 22 -- Subject: [Microscopy] Re: SEM/TEM-Buffer for a higher pH } 6, 22 -- Cc: "Microscopy Listserver" {microscopy-at-microscopy.com} } 6, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 6, 22 -- X-Scanned-By: MIMEDefang 2.51 on 152.16.199.201 } ==============================End of - Headers============================== }
==============================Original Headers============================== 5, 22 -- From Rosemary.White-at-csiro.au Thu Oct 12 16:45:35 2006 5, 22 -- Received: from act-MTAout2.csiro.au (act-MTAout2.csiro.au [150.229.7.38]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CLjX99032008 5, 22 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 16:45:34 -0500 5, 22 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=TNdkGIbxJWsUaiiJzdToDEbWNEpXyxE/knlrwvDnwxsnjF9Rrvx0u+LQVGUXNx7VCnlObtIeSpSsvtb/YeLNPmi5d0Su+/3VudyufDe/ffRD6axuQ93xMWNr/0et2Aia; 5, 22 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 5, 22 -- by act-MTAout2.csiro.au with ESMTP; 13 Oct 2006 07:44:40 +1000 5, 22 -- X-IronPort-AV: i="4.09,301,1157292000"; 5, 22 -- d="scan'208"; a="115407943:sNHT29599632" 5, 22 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 5, 22 -- Fri, 13 Oct 2006 07:45:22 +1000 5, 22 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 5, 22 -- Date: Fri, 13 Oct 2006 07:47:21 +1000 5, 22 -- Subject: Re: [Microscopy] Re: SEM/TEM-Buffer for a higher pH 5, 22 -- From: Rosemary White {Rosemary.White-at-csiro.au} 5, 22 -- To: {microscopy-at-microscopy.com} 5, 22 -- Message-ID: {C154F509.197E7%Rosemary.White-at-csiro.au} 5, 22 -- In-Reply-To: {200610121832.k9CIWIgZ024019-at-ns.microscopy.com} 5, 22 -- Mime-version: 1.0 5, 22 -- Content-type: text/plain; charset="US-ASCII" 5, 22 -- Content-transfer-encoding: 7bit 5, 22 -- X-OriginalArrivalTime: 12 Oct 2006 21:45:22.0676 (UTC) FILETIME=[B82E2B40:01C6EE47] ==============================End of - Headers==============================
Considering that plants have 'rigid' cell walls, its amazing how they achieve sometimes very rapid movement!
I came across a paper on Mimosa pudica a couple of years ago, in relation to the anatomy of the motor organs at the bases of the prtiole, rachis and pinnule. The paper focuses on the vascular tissue, but its a start:
Fleurat-Lessard and Bonnemain (1978) Structural and ultrastructural charactersitics of the vascular apparatus of the sensitive plant (Mimosa pudica L.) Protoplasma 94: 127-143
Hope this helps,
Mark
Mark Talbot CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 mark.talbot-at-csiro.au ph: 02-6246 5256 fax: 02-6246 5334
==============================Original Headers============================== 8, 30 -- From mark.talbot-at-csiro.au Thu Oct 12 17:48:11 2006 8, 30 -- Received: from act-MTAout6.csiro.au (act-MTAout6.csiro.au [150.229.7.43]) 8, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CMm9Rq020090 8, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 17:48:10 -0500 8, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=n2eEEmLInUTTCKsjzABbDUEd2aEiZMnTnOoMssiF2gFPo2LzK8F7safJNb+TdJVHWVhCM3Rcv8KCrohyKgsb0JX2YbyOoKSwWklBFmQKIqz+9NGwULOdxPlCWR9DGI1X; 8, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 8, 30 -- by act-MTAout6.csiro.au with ESMTP; 13 Oct 2006 08:47:16 +1000 8, 30 -- X-IronPort-AV: i="4.09,301,1157292000"; 8, 30 -- d="scan'208"; a="115417095:sNHT24606672" 8, 30 -- Received: from exvicn1-mel.nexus.csiro.au ([138.194.3.60]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 30 -- Fri, 13 Oct 2006 08:47:59 +1000 8, 30 -- Received: from exnswn1-syd.nexus.csiro.au ([130.155.3.31]) by exvicn1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 30 -- Fri, 13 Oct 2006 08:47:59 +1000 8, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 30 -- content-class: urn:content-classes:message 8, 30 -- MIME-Version: 1.0 8, 30 -- Content-Type: text/plain; 8, 30 -- charset="Windows-1252" 8, 30 -- Subject: Re: Mimosa questions... 8, 30 -- Date: Fri, 13 Oct 2006 08:47:58 +1000 8, 30 -- Message-ID: {BF94750921492E4AA5B825026FCE6A2401489FCE-at-exnswn1-syd.nexus.csiro.au} 8, 30 -- X-MS-Has-Attach: 8, 30 -- X-MS-TNEF-Correlator: 8, 30 -- Thread-Topic: Re: Mimosa questions... 8, 30 -- Thread-Index: AcbuTzeBN4bi+CD3Q9SYs6jcdEe4Tw== 8, 30 -- From: {mark.talbot-at-csiro.au} 8, 30 -- To: {Microscopy-at-microscopy.com} 8, 30 -- X-OriginalArrivalTime: 12 Oct 2006 22:47:59.0394 (UTC) FILETIME=[775BD820:01C6EE50] 8, 30 -- Content-Transfer-Encoding: 8bit 8, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CMm9Rq020090 ==============================End of - Headers==============================
Michael, I googled neslab and got www.neslab.com . They're apparently part of the Thermo family now (as is nearly everyone in the EM accessories industry). There were also a huge number of used Neslab chillers listed. My customers have generally had good luck with their products and I've found their service to be quite responsive and helpful (along with Haskris).
No stake, just like their stuff.
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: mdelann1-at-jhmi.edu [mailto:mdelann1-at-jhmi.edu] Sent: Wednesday, October 11, 2006 9:39 PM To: kenconverse-at-qualityimages.biz
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Email: mdelann1-at-jhmi.edu Name: Michael Delannoy
Organization: Assistant Director Imaging Facility JHSM
Title-Subject: [Filtered] chillers
Question: Hello I need a few companies who would sell used/ refurbished water chillers/recirculators. I need a supply for my LEO 1530 FESEM Does anyone know the heat capaciy required? Does anyone know the Neslabs website or contact info? Thank You M Delannoy 410 955-1365
==============================Original Headers============================== 6, 12 -- From zaluzec-at-microscopy.com Wed Oct 11 20:35:33 2006 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C1ZVoF003898 6, 12 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 20:35:33 -0500 6, 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- Message-Id: {p06110405c1534c51302b-at-[206.69.208.22]} 6, 12 -- Date: Wed, 11 Oct 2006 20:35:30 -0500 6, 12 -- To: microscopy-at-microscopy.com 6, 12 -- From: mdelann1-at-jhmi.edu (by way of MicroscopyListserver) 6, 12 --
Chillers....a really interesting area and quite surprising.
The Thermo units I think across the board have 0.5 gallon reservoirs. If your flow rate is really low, this should not be an issue. I think like .5GPH or so which is what the FEI systems use. They can do this because their systems dump heat into the area where the SEM and expansion racks are located. Zeiss takes a different approach to cooling. They use the Haskris chillers with 5 gallon reservoirs and add some additional interesting options. A key one is the hot gas bypass.
Myself and others have second-guessed the Zeiss folks and come out with the short end of the stick. The hot gas bypass is a feature that dumps hot refrigerant back into the reservoir thus causing the compressor to not shut off. The net results of this are that the compressor rarely shuts off and water temperature at high (30GPM or H, not sure) flow is constant. Also, the chiller takes most all of the SEM plinth heat out to where the chiller is. So, if located in different rooms, the SEM room is easy to cool.
Another factor is that the compressor does not short cycle. This is fatal to the compressor if it does short cycle--on and off in a short period of time. The start relay will eventually burn out and require replacement of the whole compressor unit if this happens. Short cycling happens when the temperature setting has hysterisis. E. g., 65F - 67F. When the water reaches 67F, the compressor starts and cools the water to 65F then shuts off. Depending on the heat load, this can happen very frequently. With low demand systems like FEI, the compressors see the same thing but seem to take it in stride. The Tecumseh compressors in the Haskris chillers apparently do not like this.
Anyway, Haskris has excellent customer service from my experience and the Thermo units are also good for their applications. So, I guess that it just "depends."
Disclaimer: blah, blah.
gary g.
At 04:32 PM 10/12/2006, you wrote:
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==============================Original Headers============================== 14, 20 -- From gary-at-gaugler.com Thu Oct 12 19:16:37 2006 14, 20 -- Received: from qsmtp1.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 k9D0GbVl026305 14, 20 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 19:16:37 -0500 14, 20 -- Received: (qmail 14988 invoked from network); 12 Oct 2006 17:16:32 -0700 14, 20 -- Received: by simscan 1.1.0 ppid: 14976, pid: 14986, t: 0.2404s 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 qsmtp1 with SMTP; 12 Oct 2006 17:16:32 -0700 14, 20 -- Message-Id: {7.0.1.0.2.20061012165435.0259ae00-at-gaugler.com} 14, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 20 -- Date: Thu, 12 Oct 2006 17:16:39 -0700 14, 20 -- To: kenconverse-at-qualityimages.biz 14, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 14, 20 -- Subject: Re: [Microscopy] RE: viaWWW:water chillers 14, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 14, 20 -- In-Reply-To: {200610122332.k9CNWUnj010065-at-ns.microscopy.com} 14, 20 -- References: {200610122332.k9CNWUnj010065-at-ns.microscopy.com} 14, 20 -- Mime-Version: 1.0 14, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Our newer EFTEM has been dead for a few weeks, so I got our venerable old Zeiss 10/A going. This meant loading in old film from 2001, cleaning out the darkroom, making up old (and black) D-19, and generally stepping back into the past. Amazingly, the images are great! We've been very happy with them; giddy, even! However, our stash of old film is nearly gone, and I remember the rant on this list a couple of years ago about the new, "improved" Kodak 4489. Indeed, one user bought new film (no instruction sheet included!) and new D-19 (new plastic packaging), and we are terribly unhappy with the results. I went back through the List's archives, and we tried all the tricks and patiently tried all kinds of dilutions of developer, development times, exposures, etc., all with poor results. We're not getting uneven development; our agitation seems to be fine. It seems that the response curve of the new film is more linear instead of the traditional S-shape, and we're not getting good detail in the darker blacks and whiter whites. And the film seems to have a brown cast!
I would be interested in hearing how any of you have been dealing with this, especially within the past year.
Mahalo, Tina
**************************************************************************** * 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============================== 7, 19 -- From tina-at-pbrc.hawaii.edu Thu Oct 12 19:30:41 2006 7, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9D0UfVV007093 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 19:30:41 -0500 7, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 7, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k9D0UbAk013635 7, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 14:30:38 -1000 (HST) 7, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k9D0Ua2g013632 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 14:30:37 -1000 (HST) 7, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 7, 19 -- Date: Thu, 12 Oct 2006 14:30:36 -1000 (HST) 7, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 7, 19 -- X-Sender: tina-at-halia 7, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 7, 19 -- Subject: Kodak 4489 film 7, 19 -- Message-ID: {Pine.GSO.4.21.0610121418500.13499-100000-at-halia} 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
I am doing fundamental research with a Megaview III digital camera from Olympus-SIS imaging (side-mounted camera). I find this material just fantastic. No need to observe on the fluorescent screen, with several advantages: - You can take a picture immediately: find a field, take a picture. 1 second later you have your results and can adjust if needed. - the camera being much more sensitive that your eyes on the fluorescent screen, you can reduce the dose of the beam, meaning less damage to your material and better contrast. - Your can organize your results immediately: no need to take notes, numerate your negative, calculate the magnification...
In addition to the camera comes a great software which allows you to organize very efficiently your pictures in databases and reports. The software also allows all sorts of analysis and calculations from your pictures. You want more? Well actually you get more, since the technical assistance is really efficient too!
Important things to consider are the resolution of the camera (although I wonder why a pathologist would need the highest resolution possible - it is an open question, i am not pathologist) and the horizontal transfer of the informations between the camera software and other software used in your hospital
Regards,
Stephane
----- Original Message ---- X-from: "GBurgess-at-exchange.hsc.mb.ca" {GBurgess-at-exchange.hsc.mb.ca} To: nizets2-at-yahoo.com Sent: Thursday, October 12, 2006 9:53:09 PM
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
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==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
==============================Original Headers============================== 24, 20 -- From nizets2-at-yahoo.com Fri Oct 13 03:17:53 2006 24, 20 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.91.145]) 24, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9D8Hrxn015799 24, 20 -- for {microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 03:17:53 -0500 24, 20 -- Received: (qmail 80657 invoked by uid 60001); 13 Oct 2006 08:17:53 -0000 24, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 24, 20 -- s=s1024; d=yahoo.com; 24, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:MIME-Version:Content-Type:Content-Transfer-Encoding; 24, 20 -- b=JchCkauFp84T1QQQjnrkI0BbEttrgkjOKqR/kR1Xjd72lz31qZD0EBCzIXiHEs7nqHw7WPipEGRllnCs6F0Fbii2a6BEl8xrQVxr7+hwlluXAcUPtutOzEGcoqtrJgC77da90Z+PdMfwkNuyNkKc77q0aWN4ueZLQOSzbXUKNuo= ; 24, 20 -- Message-ID: {20061013081753.80655.qmail-at-web37413.mail.mud.yahoo.com} 24, 20 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Fri, 13 Oct 2006 01:17:53 PDT 24, 20 -- Date: Fri, 13 Oct 2006 01:17:53 -0700 (PDT) 24, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 24, 20 -- Subject: Re: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 24, 20 -- To: GBurgess-at-exchange.hsc.mb.ca 24, 20 -- Cc: microscopy-at-microscopy.com 24, 20 -- MIME-Version: 1.0 24, 20 -- Content-Type: text/plain; charset=ascii 24, 20 -- Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9D8Hrxn015799 ==============================End of - Headers==============================
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Email: mark.grimson-at-ttu.edu Name: Mark Grimson
Organization: Texas Tech University
Title-Subject: [Filtered] Horizontal lines on a Gatan slow-scan TEM camera
Question: Hello. We recently recieved an H-8100 with a bottom mount Gatan slow-scan CCD cooled camera Model 794 multi-scan camera). Before it left its old location the camera was functioning normally, but when it arrived here, it developed some problems, in particular, horizontal lines running across the acquisition screen in all 3 modes. The image that is projected on the camera can be observed between the lines. Has anyone had experience with this problem before. I would appreciate any suggestions before sending the camera back to Gatan for repairs. Thanks for any advice or comments. Mark
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Email: jvtaylo-at-emory.edu Name: Jeannette Taylor
Organization: IM&MF, Emory University
Title-Subject: [Filtered] Surplus DS130 LaB6 SEM
Question: Dear All, we have a Topcon DS130 SEM, with LaB6 filament, in operating condition, available very cheap to interested parties. This instrument has in-lens imaging as well as conventional below-the-lens imaging. Other features as well. Please inquire.
Thanks, Jeannette Taylor
Jeannette Taylor, Technologist II IM&MF, Emory University (404) 712-8674 jvtaylo-at-emory.edu
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Title-Subject: [Filtered] Electron Microscopist Salary
Question: I am trying to hire an electron microscopist who has 3-5 years of research and technical experience using TEM. I am having difficulties getting past HR who want to pay this techmician far less than I think reasonable for the position. Can anyone provide me with salary ranges suggested for RA-I type positions, in a university setting, for electron microspists.
I never had any problems with the new 4489 film. I developed it in D-19, diluted 1:2, for 4 min 15 seconds at 70 F. Gentle agitation for the first 30 seconds, then for 5 seconds every 30 seconds. Rinse, fix and wash as usual. They only difference is that I make my D-19 (and all of my other B&W developers) from scratch. Not from the Kodak package, I have the formula, each of the raw ingredients and I weigh them out and dissolve them in the proper order in distilled water. I am surprised you got good results from old, black D-19!
Geoff
tina-at-pbrc.hawaii.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============================== 9, 34 -- From mcauliff-at-umdnj.edu Fri Oct 13 09:03:10 2006 9, 34 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DE39Ls020998 9, 34 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 09:03:09 -0500 9, 34 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 9, 34 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id AF22A1C328C 9, 34 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 10:03:08 -0400 (EDT) 9, 34 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 9, 34 -- by zix01.umdnj.edu (Proprietary) with ESMTP id 29B05A7B4A 9, 34 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 10:03:07 -0400 (EDT) 9, 34 -- Received: from ([130.219.34.131]) 9, 34 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.38457316; 9, 34 -- Fri, 13 Oct 2006 09:53:54 -0400 9, 34 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 9, 34 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 9, 34 -- id {0J7200B01U9ESQ-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 9, 34 -- for microscopy-at-msa.microscopy.com; Fri, 13 Oct 2006 09:53:54 -0400 (EDT) 9, 34 -- Received: from [127.0.0.1] ([10.138.2.123]) 9, 34 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 9, 34 -- 2004)) with ESMTP id {0J72008BDUKN2C-at-Polaris.umdnj.edu} ; Fri, 9, 34 -- 13 Oct 2006 09:53:12 -0400 (EDT) 9, 34 -- Date: Fri, 13 Oct 2006 09:54:22 -0400 9, 34 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 9, 34 -- Subject: Re: [Microscopy] Kodak 4489 film 9, 34 -- In-reply-to: {200610130032.k9D0WRhh009323-at-ns.microscopy.com} 9, 34 -- To: tina-at-pbrc.hawaii.edu, MicroscopyListserver {microscopy-at-msa.microscopy.com} 9, 34 -- Message-id: {452F9A8E.7050603-at-umdnj.edu} 9, 34 -- MIME-version: 1.0 9, 34 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 9, 34 -- Content-transfer-encoding: 7BIT 9, 34 -- X-Accept-Language: en-us, en 9, 34 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 9, 34 -- Gecko/20040804 Netscape/7.2 (ax) 9, 34 -- References: {200610130032.k9D0WRhh009323-at-ns.microscopy.com} ==============================End of - Headers==============================
Hi Tina, I use new improved Kodak Estar 4489. I had to change safety light in my dark room from sodium light to a red filter because this film is apparently sensitive to sodium light (according to the manufacturer insert). Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Fri Oct 13 09:43:18 2006 1, 22 -- Received: from mx2.upei.ca (magellanic.cs.upei.ca [137.149.3.22]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DEhI1t010869 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 09:43:18 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx2.upei.ca with esmtp (Exim 4.50 #1 (Debian)) 1, 22 -- id 1GYOFq-0005EI-Se 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 11:43:19 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 13 Oct 06 11:43:18 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 13 Oct 06 11:42:45 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-msa.microscopy.com 1, 22 -- Date: Fri, 13 Oct 2006 11:36:08 -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: Re: Kodak 4489 film 1, 22 -- Message-ID: {452F7A27.15586.6A703A-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
Microscopy Today magazine published a salary survey in January 2005, attached.* I would add a couple of percent to reflect salary increases between late 2004, when this data was submitted, and now. We'll be doing another survey in about six months.
Ron Anderson, Editor Microscopy Today
Subscribe at http://www.microscopy-today.com, free in North America. *Attached in response to Karen
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==============================Original Headers============================== 5, 19 -- From microscopytoday-at-tampabay.rr.com Fri Oct 13 10:00:30 2006 5, 19 -- Received: from ms-smtp-06.tampabay.rr.com (ms-smtp-06.tampabay.rr.com [65.32.5.136]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DF0Ubw021753 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Fri, 13 Oct 2006 10:00:30 -0500 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-06.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k9DF0RBf011246 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Fri, 13 Oct 2006 11:00:29 -0400 (EDT) 5, 19 -- Message-ID: {452FAA09.5050305-at-tampabay.rr.com} 5, 19 -- Date: Fri, 13 Oct 2006 11:00:25 -0400 5, 19 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] viaWWW: Electron Microscopist Salary 5, 19 -- References: {200610131354.k9DDslTx002930-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200610131354.k9DDslTx002930-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==============================
You may use any composite video monitor (9" B&W security monitor), given that H-600 takes composite video- I am not 100% certain about H-600, but most data monitors like that use standard composite video 1V p-p; 75 Ohm. Worth checking out IMHO.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com ----- Original Message ----- X-from: {john.brealey-at-imvs.sa.gov.au} To: {vitalylazar-at-att.net} Sent: Wednesday, October 11, 2006 10:03 PM
Dear Colleagues...
We're trying to process skin tissue samples for TEM. Despite several attempts with different protocols, we couldn't achieve fully satisfactory results.Has anybody experienced skin tissue processing? Are there "tips and tricks" for processing skin tissue? Thank you in advance...
Dr. Nejat Yilmaz
==============================Original Headers============================== 8, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 14:17:22 2006 8, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DJHLFG015879 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 14:17:22 -0500 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 25FF94508A 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:59 +0300 (EEST) 8, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr 8, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 8, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 8, 28 -- with ESMTP id K66xSD0aheDH for {Microscopy-at-microscopy.com} ; 8, 28 -- Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 21AA6450AE 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- To: {Microscopy-at-microscopy.com} 8, 28 -- Subject: Skin processing 8, 28 -- From: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Date: Fri, 13 Oct 2006 22:32:56 EEST 8, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- X-Priority: 3 (Normal) 8, 28 -- X-Originating-Ip: [85.104.168.63] 8, 28 -- X-Mailer: NOCC v0.9.5 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="ISO-8859-9" 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- Message-Id: {20061013193256.21AA6450AE-at-mail.mersin.edu.tr} ==============================End of - Headers==============================
What kind of skin are you processing? We do mouse skin routinely and successfully for TEM. I usually embed in Spurrs resin and do long infiltration steps.
Lesley
Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 -----Original Message----- X-from: nyilmaz-at-mersin.edu.tr [mailto:nyilmaz-at-mersin.edu.tr] Sent: Friday, October 13, 2006 3:25 PM To: lesley.bechtold-at-jax.org
Dear Colleagues...
We're trying to process skin tissue samples for TEM. Despite several attempts with different protocols, we couldn't achieve fully satisfactory results.Has anybody experienced skin tissue processing? Are there "tips and tricks" for processing skin tissue? Thank you in advance...
Dr. Nejat Yilmaz
==============================Original Headers============================== 8, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 14:17:22 2006 8, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DJHLFG015879 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 14:17:22 -0500 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 25FF94508A 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:59 +0300 (EEST) 8, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr 8, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 8, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 8, 28 -- with ESMTP id K66xSD0aheDH for {Microscopy-at-microscopy.com} ; 8, 28 -- Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 21AA6450AE 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- To: {Microscopy-at-microscopy.com} 8, 28 -- Subject: Skin processing 8, 28 -- From: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Date: Fri, 13 Oct 2006 22:32:56 EEST 8, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- X-Priority: 3 (Normal) 8, 28 -- X-Originating-Ip: [85.104.168.63] 8, 28 -- X-Mailer: NOCC v0.9.5 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="ISO-8859-9" 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- Message-Id: {20061013193256.21AA6450AE-at-mail.mersin.edu.tr} ==============================End of - Headers==============================
==============================Original Headers============================== 18, 23 -- From lesley.bechtold-at-jax.org Fri Oct 13 14:36:20 2006 18, 23 -- Received: from carmen.jax.org (carmen.jax.org [192.43.249.16]) 18, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DJaKC4026581 18, 23 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 13 Oct 2006 14:36:20 -0500 18, 23 -- Received: from jcs-mid-prod.jax.org (jcs-mid-prod.jax.org [192.43.249.134]) 18, 23 -- by carmen.jax.org (8.12.11/8.12.11) with ESMTP id k9DIUA01026242; 18, 23 -- Fri, 13 Oct 2006 15:36:14 -0400 (EDT) 18, 23 -- (envelope-from lesley.bechtold-at-jax.org) 18, 23 -- Received: from spikey.jax.org by jcs-mid-prod.jax.org 18, 23 -- with ESMTP id 129656101160768136; Fri, 13 Oct 2006 15:35:36 -0400 18, 23 -- From: "Lesley Bechtold" {lesley.bechtold-at-jax.org} 18, 23 -- To: "nyilmaz-at-mersin.edu.tr" {nyilmaz-at-mersin.edu.tr} 18, 23 -- CC: "Microscopy Network" {microscopy-at-ns.microscopy.com} 18, 23 -- Subject: RE: [Microscopy] Skin processing 18, 23 -- Date: Fri, 13 Oct 2006 15:35:35 -0400 18, 23 -- Message-ID: {20061013153535960.00000006260-at-spikey} 18, 23 -- In-Reply-To: {200610131925.k9DJPOxc023765-at-ns.microscopy.com} 18, 23 -- X-Mailer: Oracle Connector for Outlook 10.1.1.0.5 71011 (11.0.8010) 18, 23 -- X-Accept-Language: en-us, en 18, 23 -- MIME-Version: 1.0 18, 23 -- Content-Type: text/plain; charset=us-ascii 18, 23 -- Content-Transfer-Encoding: 8bit 18, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9DJaKC4026581 ==============================End of - Headers==============================
Hi Nejat I do not have a protocol for TEM of skin, but Pierre Coulombe is the master. Contact him at coulombe-at-jhmi.edu. He can get you what you need. David
On Oct 13, 2006, at 12:21 PM, nyilmaz-at-mersin.edu.tr 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... } } We're trying to process skin tissue samples for TEM. Despite several } attempts with different protocols, we couldn't } achieve fully satisfactory results.Has anybody experienced skin } tissue processing? } Are there "tips and tricks" for } processing skin tissue? } Thank you in advance... } } Dr. Nejat Yilmaz } } } } } } } ==============================Original } Headers============================== } 8, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 14:17:22 2006 } 8, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr } [193.255.128.3]) } 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k9DJHLFG015879 } 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 } 14:17:22 -0500 } 8, 28 -- Received: from localhost (localhost.mersin.edu.tr } [127.0.0.1]) } 8, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 25FF94508A } 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 } 22:32:59 +0300 (EEST) } 8, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr } 8, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) } 8, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd- } new, port 10024) } 8, 28 -- with ESMTP id K66xSD0aheDH for {Microscopy-at-microscopy.com} ; } 8, 28 -- Fri, 13 Oct 2006 22:32:56 +0300 (EEST) } 8, 28 -- Received: from localhost (localhost.mersin.edu.tr } [127.0.0.1]) } 8, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 21AA6450AE } 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 } 22:32:56 +0300 (EEST) } 8, 28 -- To: {Microscopy-at-microscopy.com} } 8, 28 -- Subject: Skin processing } 8, 28 -- From: {nyilmaz-at-mersin.edu.tr} } 8, 28 -- Date: Fri, 13 Oct 2006 22:32:56 EEST } 8, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} } 8, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} } 8, 28 -- X-Priority: 3 (Normal) } 8, 28 -- X-Originating-Ip: [85.104.168.63] } 8, 28 -- X-Mailer: NOCC v0.9.5 } 8, 28 -- Content-Type: text/plain; } 8, 28 -- charset="ISO-8859-9" } 8, 28 -- Content-Transfer-Encoding: 8bit } 8, 28 -- Message-Id: {20061013193256.21AA6450AE-at-mail.mersin.edu.tr} } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Fri Oct 13 15:19:52 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DKJo2b005573 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Oct 2006 15:19:51 -0500 5, 22 -- Received: from localhost (boromir.email.arizona.edu [10.0.0.217]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 8498911447A2 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Oct 2006 13:19:46 -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 D8574114B450 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Oct 2006 13:19:37 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200610131921.k9DJLjng021171-at-ns.microscopy.com} 5, 22 -- References: {200610131921.k9DJLjng021171-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {2623A477-CC5A-4437-B692-39EA95A4157E-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] Skin processing 5, 22 -- Date: Fri, 13 Oct 2006 13:19:33 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
anybody out there believing /considering, that there are differences between human and animal tissue ??....if not, I could send my protocol for human, diagnostic skin tissue (embedding in epoxid resin...Glycidether100/DMSA/MNA/DMP-30)...the differences for embedding method perhaps are not that big......
best regards, Wolfgang Muss SALZBURG, Austria
---------- Von: nyilmaz-at-mersin.edu.tr[SMTP:nyilmaz-at-mersin.edu.tr] Antwort an: nyilmaz-at-mersin.edu.tr Gesendet: Samstag, 14. Oktober 2006 00:12 An: W.Muss-at-salk.at Betreff: [Microscopy] Addition for skin processing
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Dear colleagues...
We are using Spurrs resin or Araldit resin and we are working on rat skins.
Thanks again
Dr. Nejat Yilmaz
==============================Original Headers============================== 9, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 17:07:02 2006 9, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DM71BN017947 9, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 17:07:01 -0500 9, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 9, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 1BEF145111 9, 28 -- for {Microscopy-at-microscopy.com} ; Sat, 14 Oct 2006 01:22:50 +0300 (EEST) 9, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr 9, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 9, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 9, 28 -- with ESMTP id HEN7dc7jBGXE for {Microscopy-at-microscopy.com} ; 9, 28 -- Sat, 14 Oct 2006 01:22:47 +0300 (EEST) 9, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 9, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 1C0A84510B 9, 28 -- for {Microscopy-at-microscopy.com} ; Sat, 14 Oct 2006 01:22:47 +0300 (EEST) 9, 28 -- To: {Microscopy-at-microscopy.com} 9, 28 -- Subject: Addition for skin processing 9, 28 -- From: {nyilmaz-at-mersin.edu.tr} 9, 28 -- Date: Sat, 14 Oct 2006 01:22:47 EEST 9, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} 9, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} 9, 28 -- X-Priority: 3 (Normal) 9, 28 -- X-Originating-Ip: [85.104.168.63] 9, 28 -- X-Mailer: NOCC v0.9.5 9, 28 -- Content-Type: text/plain; 9, 28 -- charset="ISO-8859-9" 9, 28 -- Content-Transfer-Encoding: 8bit 9, 28 -- Message-Id: {20061013222247.1C0A84510B-at-mail.mersin.edu.tr} ==============================End of - Headers==============================
==============================Original Headers============================== 19, 39 -- From W.Muss-at-salk.at Sat Oct 14 06:16:03 2006 19, 39 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9] (may be forged)) 19, 39 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9EBG2uW008724 19, 39 -- for {Microscopy-at-microscopy.com} ; Sat, 14 Oct 2006 06:16:03 -0500 19, 39 -- Received: from localhost (localhost [127.0.0.1]) 19, 39 -- by hermes.salk.at (Postfix) with ESMTP id D4B38C38A0; 19, 39 -- Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 19, 39 -- Received: from hermes.salk.at ([127.0.0.1]) 19, 39 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 19, 39 -- with ESMTP id 3uecVCrpXpyB; Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 19, 39 -- by hermes.salk.at (Postfix) with ESMTP id 59134C3886; 19, 39 -- Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- Received: from localhost (localhost [127.0.0.1]) 19, 39 -- by hermes.lks.at (Postfix) with ESMTP id 36CAC5A9020; 19, 39 -- Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- Received: from hermes.lks.at ([127.0.0.1]) 19, 39 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 19, 39 -- with ESMTP id 23523-07; Sat, 14 Oct 2006 13:15:59 +0200 (CEST) 19, 39 -- Received: from c1pa003 (unknown [192.168.42.3]) 19, 39 -- by hermes.lks.at (Postfix) with SMTP id D9A715A900A; 19, 39 -- Sat, 14 Oct 2006 13:15:59 +0200 (CEST) 19, 39 -- Received: by localhost with Microsoft MAPI; Sat, 14 Oct 2006 13:15:59 +0200 19, 39 -- Message-ID: {01C6EF92.E40C1AA0.W.Muss-at-salk.at} 19, 39 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 19, 39 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 19, 39 -- To: "'nyilmaz-at-mersin.edu.tr'" {nyilmaz-at-mersin.edu.tr} 19, 39 -- Cc: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 19, 39 -- Subject: [Microscopy] RE: Addition for skin processing 19, 39 -- Date: Sat, 14 Oct 2006 13:15:58 +0200 19, 39 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 19, 39 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 19, 39 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 19, 39 -- MIME-Version: 1.0 19, 39 -- Content-Type: text/plain; charset="us-ascii" 19, 39 -- Content-Transfer-Encoding: 7bit 19, 39 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 19, 39 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
Thanks to all who replied and gave advice. I'll forward the info to our electrician. BTW, our CRT screen came on for 10 minutes this morning (Monday - the TEM is still relatively cold).
John Brealey
Quote... "I think we have the same scope of the same vintage. Likewise, over the past 2 yrs the CRT has had problems. Unusually long warm up times are the norm. It was about 10min...now it's up to 30 min before the CRT kicks in. After it warms up however..it has not been a problem at all. What I don't understand is how you operate blindly to set up numbers etc. It seems that if the CRT is not on, we cannot even get a beam. I've tried to run blind with it only a few times...maybe I'll have to do it again one day. I hope not. We are still under the Hitachi service contract, and it seems that every time they come over they try something new. They have swapped out the CRT and some electrical component boards. Nothing has worked thus far. I'll be interested in any real solutions that you can share as to using a stand alone monitor. Good luck. Linda"
Linda, we have the opposite problem to you - our CRT comes on when cold then disappears after about 10 minutes. I suspect we have a faulty wire somewhere or a fine crack on a circuit board that expands as it heats up. Despite the lack of illumination on the CRT screen, all functions performed through the CRT keypad are working. Your problem sounds different - maybe a power supply problem.
Quote... "You may use any composite video monitor (9" B&W security monitor), given that H-600 takes composite video- I am not 100% certain about H-600, but most data monitors like that use standard composite video 1V p-p; 75 Ohm. Worth checking out IMHO. Vitaly Feingold"
Thanks for the info Vitaly.
Quote... "Try to replace all the capacitors in the circuit board. It is possible the high voltage is not working for the crt."
Thanks Ray.
Quote... "It sounds like a heat related issue. I had to remember an old trick for a problem we recently had. I turned a duster can upside down and sprayed liquid around at some suspect components. When I sprayed the bad one, the unit snapped back to life momentarily. I could then replace the bad component on the first attempt. I would be careful around a CRT and wouldn't be chilling the glass lest something break, but I suspect something in the driver electronics has gone bad. My next recourse would be to look for a replacement CRT from an old scope. I've had to do that with two different EDS units. Warren"
Thanks Warren.
==============================Original Headers============================== 12, 33 -- From john.brealey-at-imvs.sa.gov.au Sun Oct 15 21:54:59 2006 12, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 12, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9G2swsr006822 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 15 Oct 2006 21:54:59 -0500 12, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 12, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9G2svJZ005767 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:57 +0930 (CST)' 12, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 12, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9G2svmJ005764 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:57 +0930 (CST)' 12, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 12, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id 25DD62EC28F 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:57 +0930 (CST) 12, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 12, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 12, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 12, 33 -- with LMTP id 3fD+Ds9W6LTn for {Microscopy-at-MSA.Microscopy.Com} ; 12, 33 -- Mon, 16 Oct 2006 12:24:42 +0930 (CST) 12, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 12, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 6DC882EC294 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:42 +0930 (CST) 12, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 12, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 12, 33 -- Subject: Hitachi H-600 CRT Problem Summary 12, 33 -- Date: Mon, 16 Oct 2006 12:24:41 +0930 12, 33 -- Message-ID: {000601c6f0ce$6ded1120$c88a140a-at-iqe36042} 12, 33 -- MIME-Version: 1.0 12, 33 -- Content-Type: text/plain; 12, 33 -- charset="us-ascii" 12, 33 -- Content-Transfer-Encoding: 7bit 12, 33 -- X-Mailer: Microsoft Office Outlook 11 12, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 12, 33 -- Thread-Index: AcbwzlUpp5RNZK5yS5m/ESEwLpfxWw== ==============================End of - Headers==============================
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I am wondering what other multi-user centers use as benchmarks when justifying hiring additional staff. I can provide total hours of equipment usage and how many extra hours and weekends I am working but I am thinking there might be something more concrete I could use. Does any one have suggestions or gone through this recently?
Best Regards,
Kirk
Kirk J. Czymmek, Ph.D. Associate Professor Department of Biological Sciences Delaware Biotechnology Institute Bio-Imaging Center, Director University of Delaware Newark, DE 19711
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 annamaria.pisi as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: annamaria.pisi Name: annamaria pisi
Organization: University
Title-Subject: [Filtered] TEM and SEM fixation
Question: I have to treat human stomach biopsies that had been frozen in liquid nitrogen and kept at -85ƒC without any protectant device. I have to fix and embedd some of them for TEM and some for SEM. What should I do? I never had samples like these? I have a traditional ultramicrotome and no cryo devices. Please help thank you annamaria
Annamaria, If these were simply plunged into liquid nitrogen to freeze them, the ultrastructure will be virtually destroyed by ice crystal growth. You might check how they were frozen before you proceed to see if it's worth it. That said you can freeze-substitute them then bring them to room temperature and embed as normal without cryodevices. Get a small Styrofoam cooler (the kind chemicals are shipped in) and some dry ice and you are in business.
A good substitution medium is 1% Osmium tetroxide in dry acetone (dissolve from the crystals). Add about 1 ml of substitution medium to 2 ml cryovials, cap the vials and freeze them upright in liquid nitrogen. Transfer your samples to the top of the frozen substitution medium in a liquid nitrogen bath and loosely cap the vials, making sure there is no liquid nitrogen inside. For the substitution chamber, line a small Styrofoam cooler with some foil then put 2 beakers in, one inside the other. Add acetone to each beaker to about 1/3 full then chill everything by surrounding the beakers with dry ice. Put a few chunks in the acetone, too. Once it's all cooled add your samples with substitution medium then top up the cooler with dry ice and put the lid on. Usually a small cooler holds about 5 pounds of dry ice which will last 2 to 5 days depending on the outside temperature, but this should be enough to complete the substitution. Once the samples warm up to room temperature they are already fixed, dehydrated and ready to embed in the resin of your choice (or to CPD and put in the SEM). Don't forget to rinse a few times in pure acetone to remove the excess Osmium.
Let me know if you need more information. Kim
annamaria.pisi-at-ns.microscopy.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/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 annamaria.pisi as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: annamaria.pisi } Name: annamaria pisi } } Organization: University } } Title-Subject: [Filtered] TEM and SEM fixation } } Question: I have to treat human stomach biopsies that had been frozen in liquid nitrogen and kept at -85ƒC without any protectant device. I have to fix and embedd some of them for TEM and some for SEM. What should I do? I never had samples like these? I have a traditional ultramicrotome and no cryo devices. Please help } thank you annamaria }
-- Kim Rensing Ph.D. Manager, Microscopy and Imaging Facility, University of Calgary, Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1 O: 403-220-3488 F: 403-270-8928
==============================Original Headers============================== 6, 27 -- From krensing-at-ucalgary.ca Mon Oct 16 09:47:19 2006 6, 27 -- Received: from mr2.ucalgary.ca (mr2.ucalgary.ca [136.159.34.166]) 6, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GElIPa026874 6, 27 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 09:47:18 -0500 6, 27 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 6, 27 -- by mr2.ucalgary.ca (Postfix) with ESMTP id 4D115366A5; 6, 27 -- Mon, 16 Oct 2006 08:47:12 -0600 (MDT) 6, 27 -- Received: from [192.168.0.100] ([136.159.164.171]) 6, 27 -- (authenticated (0 bits)) 6, 27 -- by smtp2.ucalgary.ca (8.11.7/8.11.6) with ESMTP id k9GEl8013753 6, 27 -- (using TLSv1/SSLv3 with cipher DHE-RSA-AES256-SHA (256 bits) verified NO); 6, 27 -- Mon, 16 Oct 2006 08:47:08 -0600 6, 27 -- Message-ID: {45339B66.5070806-at-ucalgary.ca} 6, 27 -- Date: Mon, 16 Oct 2006 08:47:02 -0600 6, 27 -- From: Kim Rensing {krensing-at-ucalgary.ca} 6, 27 -- Organization: Microscopy and Imaging Facility, U. of Calgary 6, 27 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 6, 27 -- MIME-Version: 1.0 6, 27 -- To: annamaria.pisi-at-ns.microscopy.com, microscopy-at-microscopy.com 6, 27 -- Subject: Re: [Microscopy] viaWWW: TEM and SEM fixation human stomach 6, 27 -- References: {200610161420.k9GEKwOR025761-at-ns.microscopy.com} 6, 27 -- In-Reply-To: {200610161420.k9GEKwOR025761-at-ns.microscopy.com} 6, 27 -- Content-Type: text/plain; charset=windows-1252; format=flowed 6, 27 -- Content-Transfer-Encoding: 8bit 6, 27 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 6, 27 -- X-UCalgary-MailScanner: Found to be clean 6, 27 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
I am sure this is an easy one - I need to trace DNA (or RNA) molecules on a digital micrograph to measure their length. Any particular program recommended? NIH Image should do it? Or something maybe better (preferentially free/cheap)?
Thanks,
Michal
==============================Original Headers============================== 5, 17 -- From Michal.Jarnik-at-fccc.edu Mon Oct 16 10:15:06 2006 5, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GFF6Qn005514 5, 17 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 10:15:06 -0500 5, 17 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) 5, 17 -- (authenticated bits=0) 5, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id k9GFF5oP020664 5, 17 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 11:15:05 -0400 (EDT) 5, 17 -- Message-ID: {4533A1F3.90404-at-fccc.edu} 5, 17 -- Date: Mon, 16 Oct 2006 11:14:59 -0400 5, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} 5, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) 5, 17 -- MIME-Version: 1.0 5, 17 -- To: microscopy-at-microscopy.com 5, 17 -- Subject: Measuring length of DNA/RNA 5, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am posting this question for a friend who is working on yeast using the following (very odd) protocol:
"These yeast are chemically fixed in PB containing 3% paraformaldehyde and 0.5% gluteraldehyde 2 h room temp. Washed twice in PB, placed in 1% sodium meta periodate, rinsed PB, quenched for 30 min in 50 mM NH4Cl in PB, rinsed PB, dehydrated in ETOH 30%, 70%, 95%, and 3x 100%, microwaved 40s, 250W. Allowed to drain dry between each 100% ETOH. Placed in LR White and microwaved 3 min, 250W, placed 4 C overnight, with two more LR White changes, then placed in LE white in embedding capsules at 60 C overnight."
Go to http://www.emc.missouri.edu/lookatthis.htm to see these yeasty beasties.
She is looking for opinions on why the cell membranes are bumpy (I said drying out between 100% ETOH changes), why the vacuoles look so strange, and why the mottled appearance in the background (water in the formvar, she asks?). When reading the above protocol again, note that these samples are NOT destined for immunolabeling.
I can't wait to see the replies to this one!
Rainy day in Columbia. How's Hawaii, Tina?
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 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 On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Mon Oct 16 13:01:51 2006 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GI1ohB019704 10, 23 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 13:01:51 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="US-ASCII" 10, 23 -- Subject: Evil protocol? 10, 23 -- Date: Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68AEE-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Evil protocol? 10, 23 -- Thread-Index: AcbxTSdHxrhXluLORD+Z8CZucccncA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 16 Oct 2006 18:01:49.0844 (UTC) FILETIME=[27296D40:01C6F14D] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GI1ohB019704 ==============================End of - Headers==============================
1. I would recommend she use glutaraldehyde instead of gluteraldehyde :) 2. Why is she doing periodate? try skipping this step to see if it impacts the morphology. 3. Why NH4Cl if she isn't going to do immuno? 4. It is hard for me to judge a microwave dehydration protocol but try a conventional non-microwave approach. I would be worried about shrinkage. I agree the drying out step is highly suspect. 5. Why no osmium? 6. Why not use your fancy cryofixation device? - Studer has published gorgeous pictures of cryo-fixed yeast. 7. If no immuno, why not a better resin (epon-substitute or epon-araldite)? 8. Tina probably would need a battery operated computer to get your message!
At 01:03 PM 10/16/06, you wrote:
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My weather comment to Tina was made while having a total blankout on the recent earthquake in Hawaii. It was definitely not meant as a bad joke at the expense of the people who went though it! My apologies if it came across like that. My bad all the way.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 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 On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 5, 23 -- From TindallR-at-missouri.edu Mon Oct 16 13:40:47 2006 5, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GIekmw009224 5, 23 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 13:40:47 -0500 5, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 5, 23 -- Mon, 16 Oct 2006 13:40:43 -0500 5, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 23 -- Content-class: urn:content-classes:message 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; 5, 23 -- charset="US-ASCII" 5, 23 -- Subject: Evil protocol oops 5, 23 -- Date: Mon, 16 Oct 2006 13:40:43 -0500 5, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68AF3-at-UM-XMAIL08.um.umsystem.edu} 5, 23 -- X-MS-Has-Attach: 5, 23 -- X-MS-TNEF-Correlator: 5, 23 -- Thread-Topic: Evil protocol oops 5, 23 -- Thread-Index: AcbxUpYLx8afvMr3S+mX6Hj+L8ADQA== 5, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- X-OriginalArrivalTime: 16 Oct 2006 18:40:43.0432 (UTC) FILETIME=[9616B280:01C6F152] 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GIekmw009224 ==============================End of - Headers==============================
Dear Listers, I would appreciate an advice on processing Drosophila mutant brain for TEM. I cannot use OsO4 or DMSO, and not sure which cryofixation and cryosubstitution approach would work the best. Thank you, Albina
-- MIKHAYLOVA,ALBINA, PhD
==============================Original Headers============================== 4, 21 -- From amich-at-ufl.edu Mon Oct 16 15:45:41 2006 4, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GKjfTb022358 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 15:45:41 -0500 4, 21 -- Received: from osgjas03.cns.ufl.edu (osgjas03.cns.ufl.edu [128.227.74.133]) 4, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k9GKjZLI5255366 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 16:45:35 -0400 4, 21 -- Message-ID: {49084854.20371161031535193.JavaMail.osg-at-osgjas03.cns.ufl.edu} 4, 21 -- Date: Mon, 16 Oct 2006 16:45:35 -0400 (EDT) 4, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 4, 21 -- To: microscopy-at-microscopy.com 4, 21 -- Subject: Drosophila brain for TEM 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 4, 21 -- Content-Transfer-Encoding: 7bit 4, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 4, 21 -- X-Originating-IP: 72.155.91.89 [72.155.91.89] 4, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
1. The protocol is odd, if used for ultrastructure purpose. It is NOT odd at all if it is used for immunolabeling.
2. The "bumpy" cell membranes (looks to me the cell walls) may actually have something to do with the cell culture, rather than EM prep - some cells (the ones with bumpy cell walls) are mostly likely dying. It also explains, in part, the 'strangeness' of the vacuoles.
3. The mottled appearance in the background, if is from the formvar, could be easily checked out by examining the formvar on the grid (no sections).
4. The following is simple method for yeast TEM ultrastructure (especially if you want to avoid osmium). It works really well -
1). Harvested yeast cells are fixed first with 2.5% (v/v) glutaraldehyde in PBS for 40 min at room temperature. 2). Rinse with PBS 3X 3). Cells are further fixed with 2% potassium permanganate in water for 1 h at room temperature. 4). Rinse with water 5). Dehydration as usual 6). Use resin other than LRW for embedding
You can check some images at http://www.uwyo.edu/Microscopy/yatlas/default.htm
Zhaojie Zhang Director, Microscopy Core Facility University of Wyoming Laramie, WY 82072 http://www.uwyo.edu/microscopy
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Monday, October 16, 2006 11:12 AM To: Z.J. Zhang
Dear Listers,
I am posting this question for a friend who is working on yeast using the following (very odd) protocol:
"These yeast are chemically fixed in PB containing 3% paraformaldehyde and 0.5% gluteraldehyde 2 h room temp. Washed twice in PB, placed in 1% sodium meta periodate, rinsed PB, quenched for 30 min in 50 mM NH4Cl in PB, rinsed PB, dehydrated in ETOH 30%, 70%, 95%, and 3x 100%, microwaved 40s, 250W. Allowed to drain dry between each 100% ETOH. Placed in LR White and microwaved 3 min, 250W, placed 4 C overnight, with two more LR White changes, then placed in LE white in embedding capsules at 60 C overnight."
Go to http://www.emc.missouri.edu/lookatthis.htm to see these yeasty beasties.
She is looking for opinions on why the cell membranes are bumpy (I said drying out between 100% ETOH changes), why the vacuoles look so strange, and why the mottled appearance in the background (water in the formvar, she asks?). When reading the above protocol again, note that these samples are NOT destined for immunolabeling.
I can't wait to see the replies to this one!
Rainy day in Columbia. How's Hawaii, Tina?
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 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 On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Mon Oct 16 13:01:51 2006 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GI1ohB019704 10, 23 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 13:01:51 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="US-ASCII" 10, 23 -- Subject: Evil protocol? 10, 23 -- Date: Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68AEE-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Evil protocol? 10, 23 -- Thread-Index: AcbxTSdHxrhXluLORD+Z8CZucccncA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 16 Oct 2006 18:01:49.0844 (UTC) FILETIME=[27296D40:01C6F14D] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GI1ohB019704 ==============================End of - Headers==============================
==============================Original Headers============================== 26, 28 -- From ZZhang-at-uwyo.edu Mon Oct 16 17:52:22 2006 26, 28 -- Received: from ninemile.uwyo.edu (ninemile.uwyo.edu [129.72.10.29]) 26, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GMqMJJ002908 26, 28 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 17:52:22 -0500 26, 28 -- Received: from UWMAIL.uwyo.edu (uwmail.uwyo.edu [172.26.4.76]) 26, 28 -- by ninemile.uwyo.edu (8.12.11.20060614/8.12.11) with ESMTP id k9GMqDI2003710; 26, 28 -- Mon, 16 Oct 2006 16:52:20 -0600 (MDT) 26, 28 -- (envelope-from ZZhang-at-uwyo.edu) 26, 28 -- Received: from TELEGRAPH5.uwyo.edu ([10.84.60.120]) by UWMAIL.uwyo.edu with Microsoft SMTPSVC(6.0.3790.211); 26, 28 -- Mon, 16 Oct 2006 16:52:05 -0600 26, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 26, 28 -- Content-class: urn:content-classes:message 26, 28 -- MIME-Version: 1.0 26, 28 -- Content-Type: text/plain; 26, 28 -- charset="us-ascii" 26, 28 -- Subject: RE: [Microscopy] Evil protocol? 26, 28 -- Date: Mon, 16 Oct 2006 16:52:04 -0600 26, 28 -- Message-ID: {C9C1AF307F12AF4087DEF87CB0E6A4ADF65D33-at-TELEGRAPH5.uwyo.edu} 26, 28 -- In-Reply-To: {200610161811.k9GIBloX027956-at-ns.microscopy.com} 26, 28 -- X-MS-Has-Attach: 26, 28 -- X-MS-TNEF-Correlator: 26, 28 -- Thread-Topic: [Microscopy] Evil protocol? 26, 28 -- Thread-Index: AcbxTptO9VHFTH6mQDezomn3LaTRFAAEBQ8Q 26, 28 -- From: "Z.J. Zhang" {ZZhang-at-uwyo.edu} 26, 28 -- To: {TindallR-at-missouri.edu} , {Microscopy-at-microscopy.com} 26, 28 -- X-OriginalArrivalTime: 16 Oct 2006 22:52:05.0137 (UTC) FILETIME=[B37C4010:01C6F175] 26, 28 -- Content-Transfer-Encoding: 8bit 26, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GMqMJJ002908 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both robert.clark-at-sharp.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: robert.clark-at-sharp.com Name: Rob
Organization: Sharp HealthCare
Title-Subject: [Filtered] TEM-Propylene Oxide v. Acetone
Question: Hi, What are the opions on using propylene oxide v. acetone mixed with resin in the final steps of resin infiltration of TEM processing? We use P.O. and have for years. The question came up as I am processing a piece of rubber tubing and am concerned the P.O. may eat away at it?
Used to be a service engineer for Hitachi now full time at U of Kentucky. I also refurbish electron microscopes for resale. I always loved the 600. I have been accumulating 600s for parts and probably have a crt around. I know I have the CRT I/O circuit board.
Couple thoughts on this if you are interested. Not certain on your problem but I suspect something with the CRT circuit unit board housed with the CRT itself. May be something with the CRT I/O board that is with all the other boards in the rack, lower left.
If you have access to a competent electronics guy especially with an oscilloscope you might try some things. On the CRT I/O board there are a couple test points; HD (horizontal drive) and VD (vertical drive). You can look at these. The schematics should tell you the frequencies. I doubt these are the problem however. Best place to look next is where the plug is to the board behind the CRT. Pain to remove the covers to get there. Take the grey large cover off, then the copper colored shroud off over the CRT. Need a long plus screwdriver for this. At the plug you will have HD pin 6, VD pin 9, Video pin 8, and +15 VDC pin 7. Pin 10 is ground. That suggestion of spraying duster upside down is a good idea once the trouble occurs. I would try and look at these when working fine and then again when it quits. I guess you have ten minutes. If you lose the +15 that will definitely cause this. If all these look OK you may be losing the +10KV to the CRT tube? Be careful around the plug and wire as it will wake you up. I would make sure the display power is off when messing with this plug. Even then the CRT will hold a charge. Once unplugged make sure and ground the tube. Usually a lot of carbon build up around the plug. Possible it is finding a way to ground. I would clean this area and the rubber cover. If your electronics guy has a high voltage probe and knows how not to get killed or short things out you should get ~8 to 10KV there when powered up. Don't think it will be right at 10KV if I remember right. Sometimes the wire to this pug and CRT may break down.
One clue to eliminate HD and/or VD is while running, visually inspect the plug end of the tube. Dark room you should some illumination on the grids in the glass.
Good luck. If I can help let me know.
Joel McClintock"
Thanks Joel, great info here. I'll forward this to our electrician - he's very good with oscilloscopes and stuff.
Quote...
"Hi John,
If you cant find a solution I would talk to these guys. SMH ELECTRONICS CO 508-291-7447 I had a monitor on an old Kevex unit that had a rolling line in it. They fixed it for a flat fee and it has been fine ever since. They specialize in component level repair of ancient electronics. They keep all the stuff from the 60's going for the FAA. They can probably diagnose and fix whatever you have.
Tom Kaye"
Thanks Tom.
==============================Original Headers============================== 17, 33 -- From john.brealey-at-imvs.sa.gov.au Mon Oct 16 19:07:01 2006 17, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 17, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9H06x5T003028 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 19:07:00 -0500 17, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 17, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9H06xJZ027770 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:36:59 +0930 (CST)' 17, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 17, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9H06xmJ027767 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:36:59 +0930 (CST)' 17, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 17, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id E5927304501 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:36:58 +0930 (CST) 17, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 17, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 17, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 17, 33 -- with LMTP id YL04FuXLCNih for {Microscopy-at-MSA.Microscopy.Com} ; 17, 33 -- Tue, 17 Oct 2006 09:36:41 +0930 (CST) 17, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 17, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id CF52C3044FE 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:35:15 +0930 (CST) 17, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 17, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 17, 33 -- Subject: Hitachi H-600 CRT 17, 33 -- Date: Tue, 17 Oct 2006 09:35:14 +0930 17, 33 -- Message-ID: {000c01c6f17f$ec79be60$c88a140a-at-iqe36042} 17, 33 -- MIME-Version: 1.0 17, 33 -- Content-Type: text/plain; 17, 33 -- charset="us-ascii" 17, 33 -- Content-Transfer-Encoding: 7bit 17, 33 -- X-Mailer: Microsoft Office Outlook 11 17, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 17, 33 -- Thread-Index: Acbxf9OxNYQW5AZXRfGvJCmDebNFoQ== ==============================End of - Headers==============================
On 16 Oct 2006 at 19:00, bozhilov-at-ucr.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 } ---------------------------------------------------------------------- } ------ } } Can a photomultiplier operate directly in the vacuum of the specimen } chamber of a regular SEM, without being enclosed in a vacuum tube? } } } Krassimir Bozhilov } }
My guess, for the little that it may be worth, is that if the regular SEM used a diffusion pump, the PMT photocathode and the dynodes would soon become too oily to emit photoelectrons.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 7, 27 -- From r.sims-at-auckland.ac.nz Mon Oct 16 19:11:13 2006 7, 27 -- Received: from harpo.itss.auckland.ac.nz (mailhost.auckland.ac.nz [130.216.190.13]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9H0BCCK012941 7, 27 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 19:11:13 -0500 7, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 7, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 21E7034A1B; 7, 27 -- Tue, 17 Oct 2006 13:11:12 +1300 (NZDT) 7, 27 -- Received: from harpo.itss.auckland.ac.nz ([127.0.0.1]) 7, 27 -- by localhost (smtpc.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 7, 27 -- with ESMTP id 03955-11; Tue, 17 Oct 2006 13:11:12 +1300 (NZDT) 7, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 7, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 020AB3453E; 7, 27 -- Tue, 17 Oct 2006 13:11:11 +1300 (NZDT) 7, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 7, 27 -- To: bozhilov-at-ucr.edu 7, 27 -- Date: Tue, 17 Oct 2006 13:15:56 +1300 7, 27 -- MIME-Version: 1.0 7, 27 -- Subject: Re: [Microscopy] PMT 7, 27 -- Cc: microscopy-at-microscopy.com 7, 27 -- Message-ID: {4534D78C.1885.11DAE85-at-localhost} 7, 27 -- Priority: normal 7, 27 -- In-reply-to: {200610170000.k9H007KA026517-at-ns.microscopy.com} 7, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 7, 27 -- Content-type: text/plain; charset=US-ASCII 7, 27 -- Content-transfer-encoding: 7BIT 7, 27 -- Content-description: Mail message body 7, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
A channel PMT might work in your situation. It eliminates the dynodes and voltage dropping resistors.
What type of application would you have in mind?
gary g.
At 05:00 PM 10/16/2006, you wrote:
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==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Mon Oct 16 19:17:19 2006 13, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9H0HJG4024184 13, 20 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 19:17:19 -0500 13, 20 -- Received: (qmail 18999 invoked from network); 16 Oct 2006 17:17:18 -0700 13, 20 -- Received: by simscan 1.1.0 ppid: 18991, pid: 18993, t: 0.1493s 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 qsmtp2 with SMTP; 16 Oct 2006 17:17:18 -0700 13, 20 -- Message-Id: {7.0.1.0.2.20061016170953.02585ea8-at-gaugler.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 20 -- Date: Mon, 16 Oct 2006 17:17:59 -0700 13, 20 -- To: bozhilov-at-ucr.edu 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] PMT 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200610170000.k9H00igN027809-at-ns.microscopy.com} 13, 20 -- References: {200610170000.k9H00igN027809-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I’m passing this on to the microscopy list with some comments.
I forgot to mention that we have a CM120 TWIN, which has a Cs of 2.2 mm, whereas your Biotwin has a Cs around 6 mm. That might explain the difference since Cs is one of the parameters that influence the envelope of the CTF.
On the other hand I don’t think an information limit of 20 Å is good enough even for a biotwin. My suggestion is to make the convergence angle as small as possible (sacrificing illumination intensity) by spreading the beam over a large specimen area. Maybe you have to use a larger spot to compensate for the loss of intensity.
I checked changing only Cs (between 2 and 6 mm) in CTF-explorer (http://clik.to/ctfexplorer) and it didn’t seem to affect the envelope very much, whereas the convergence angle certainly does.
I wonder if we’ll get any comments on this from the rest of the list.
Philip
Philip Koeck Karolinska Institute and Södertörns Högskola Dept. of Bioscience at Novum tel: +46 8 608 9186 fax: +46 8 608 9290 web: http://www.csb.ki.se/users/philip/philown.html ________________________________________ X-from: Pan Xijiang [mailto:panxijiang-at-gmail.com] Sent: 16 October 2006 18:16 To: Philip.Koeck
Hi Rob,
We have switched almost entirely to acetone for our dehydrating and infiltration steps. It works just fine with our variious permutations of Epon (EMBed), Spuur's, and Araldite and allows us to completely avoid the carcinogenic and hard-to-handle PO steps. We also use it occasionally as a transition between ethanol and resin in the rare cases when we want most of the dehydration done in the somewhat less-extractive ethanol. We use microwave processing for the vast majority of our samples, but it also works with longer procedures.
We have no experience with embedding rubber tubing, however.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 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 On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: robert.clark-at-sharp.com [mailto:robert.clark-at-sharp.com] Sent: Monday, October 16, 2006 6:48 PM To: Tindall, Randy D.
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Email: robert.clark-at-sharp.com Name: Rob
Organization: Sharp HealthCare
Title-Subject: [Filtered] TEM-Propylene Oxide v. Acetone
Question: Hi, What are the opions on using propylene oxide v. acetone mixed with resin in the final steps of resin infiltration of TEM processing? We use P.O. and have for years. The question came up as I am processing a piece of rubber tubing and am concerned the P.O. may eat away at it?
Since you are asking it sounds like you have a problem. Having worked with a 100CX and 100S (23 yrs old and still going strong), the answer would have to be well in excess of 3000 plates for either one.
Two things seem to cause plate jams (in JEOL's): (1) plates loaded backwards in the film supply box (i.e. the notch on the wrong side, the boxes have just enough play in them that the weight of ~ 10 plate is enough to force a backwards plate at the bottom into the box. (2) bent plates.
Good luck.
On 11 Oct 2006 at 16:07, rcsencsits-at-lbl.gov 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 JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548 } } } } } ==============================Original Headers============================== } 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 } 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) } 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 } 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) } 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; } 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) } 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) } 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} } 7, 21 -- Content-Transfer-Encoding: 7bit } 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} } 7, 21 -- Subject: JEOL camera question } 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 } 7, 21 -- To: Microscopy-at-Microscopy.Com } 7, 21 -- X-Mailer: Apple Mail (2.752.3) } 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 } 7, 21 -- X-Virus-Status: Clean } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 10, 27 -- From edelmare-at-muohio.edu Tue Oct 17 12:23:22 2006 10, 27 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HHNMtk009258 10, 27 -- for {microscopy-at-Microscopy.com} ; Tue, 17 Oct 2006 12:23:22 -0500 10, 27 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 10, 27 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9HHNJcO014117; 10, 27 -- Tue, 17 Oct 2006 13:23:20 -0400 10, 27 -- Received: from [192.168.1.23] ([134.53.14.97]) 10, 27 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9HHNI9X019832; 10, 27 -- Tue, 17 Oct 2006 13:23:19 -0400 10, 27 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 10, 27 -- To: rcsencsits-at-lbl.gov 10, 27 -- Date: Tue, 17 Oct 2006 13:23:31 -0400 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] JEOL camera question 10, 27 -- CC: microscopy-at-Microscopy.com 10, 27 -- Message-ID: {4534D953.29753.193DDC48-at-edelmare.muohio.edu} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200610112107.k9BL7fRi029824-at-ns.microscopy.com} 10, 27 -- References: {200610112107.k9BL7fRi029824-at-ns.microscopy.com} 10, 27 -- X-mailer: Pegasus Mail for Windows (4.41) 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-Real-ConnectIP: 134.53.14.97 10, 27 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 10, 27 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 ==============================End of - Headers==============================
For a long time at CMU we tried to get a Digital System for our TEM. But out limitation was the lack of a 35mm port on the CM10. I didn't see that as a starting point for analysis.
#1 do you have a 35mm port? If so that's great. If not, open the wallet. As Mike mentioned the bottom mount camera systems typically are much more money.
(before more points another perspective)
Here in our facility we have a digital camera on our Philips 410. It has a 1k x 1k AMT (and no I have no financial interest in any vendors microscope or camera). The contrast and sensitivity means fast screening of images with intensities too low to visualize on the old 'analog' screen. The down side for us is the pixel size. Fewer pixels can mean increased sensitivity and decreased exposure times (faster live rates), but it means you are limited to on screen magnification. That has been the biggest challenge for me switching from film.
Film is great - take the image, and you have plenty of magnification overhead (enlarging). With the 1024x1024 camera, the image that you collect needs to be at the ultimate magnification. The sensitivity of the camera is such that the lower intensities work fine, but, goes counter the old adage of: take the image at the lowest magnification possible with the beam as far from cross over as possible.
With free software like autostich (google it) doing a montage is quite simple and direct (if you work with 8 bit files), but not quite the same as having a 2k or 4k array to work with. Even an automated montage system (assuming you have an automated stage that the camera software can integrate with) can overcome the limitations of few pixels, but potentially can add cost or exposure time, an still is not (in my limited opinion) quite as good as having the image collected on one array (film or CCD).
Last few thoughts to think about...
#2 How do you deal with the negatives: Routinely enlarge?
#3 Final print size (is a 1k x 1k image enough pixels)
#4 Speed and exposure times (goes to what Gary mentioned)
Good luck, and I hope that all makes sense (and I hope I'm not drastically repeating what others have already mentioned).
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: GBurgess-at-exchange.hsc.mb.ca [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, October 12, 2006 3:50 PM To: Williams, Geoffrey
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
==============================Original Headers============================== 29, 30 -- From Geoffrey_Williams-at-brown.edu Tue Oct 17 14:43:44 2006 29, 30 -- Received: from draco.services.brown.edu (draco.services.brown.edu [128.148.106.172]) 29, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HJhiR6022478 29, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:43:44 -0500 29, 30 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 29, 30 -- by draco.services.brown.edu (Switch-3.1.10/Switch-3.1.7/) with ESMTP id k9HJhhiR015046 29, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 15:43:43 -0400 (EDT) 29, 30 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 29, 30 -- Tue, 17 Oct 2006 15:43:43 -0400 29, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 29, 30 -- Content-class: urn:content-classes:message 29, 30 -- MIME-Version: 1.0 29, 30 -- Content-Type: text/plain; 29, 30 -- charset="us-ascii" 29, 30 -- Subject: RE: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 29, 30 -- Date: Tue, 17 Oct 2006 15:43:42 -0400 29, 30 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F04ACAF61-at-MAIL1.AD.Brown.Edu} 29, 30 -- In-Reply-To: {200610121950.k9CJoHMl005823-at-ns.microscopy.com} 29, 30 -- X-MS-Has-Attach: 29, 30 -- X-MS-TNEF-Correlator: 29, 30 -- Thread-Topic: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 29, 30 -- Thread-Index: AcbuN6X4PVQoLM5RQP25bN9/NOU+/gD6knpA 29, 30 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 29, 30 -- To: {Microscopy-at-microscopy.com} 29, 30 -- X-OriginalArrivalTime: 17 Oct 2006 19:43:43.0588 (UTC) FILETIME=[8DA67A40:01C6F224] 29, 30 -- X-Brown-Proofpoint: Not Infected 29, 30 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0609290000 definitions=main-0610170010 29, 30 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0609290000 definitions=main-0610170010 29, 30 -- Content-Transfer-Encoding: 8bit 29, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9HJhiR6022478 ==============================End of - Headers==============================
Our lab is disposing of a JEOL 2000FX TEM/STEM (see specs below). Please contact me off-line if you are interested in this instrument.
Specifications for JEOL 2000FX TEM Model # JEOL 2000FX TEM/STEM Serial # EM133042-32
Capabilities and features Purchased in 1987 Excellent condition; maintained by JEOL personnel; no serious outstanding or recurring problems 40 kV to 200 kV LaB6 gun Diffusion and ion pumped column and camera Specimen goniometer with 60 travel STEM capability using Polaroid film or 35-mm image acquisition Cold finger for sample EDS compatible TEM is not RS232 capable Air compressor to run pneumatic valves is not available (runs on house nitrogen in our facility) Gatan digital image acquisition system (see below) Equipment for plate camera (see below) Haskris R100 water-cooled chiller (1 year old; uses R134a coolant) Stand-alone film desiccator (without mechanical pump) JEOL speciality tools, includes hoist for gun assembly Documentation
Cameras and detectors: Gatan MSC-794 digital camera, 1024 x 1024 pixel CCD, lower mount (below plate camera); computer, monitor and software not included Polaroid film/CRT imaging system for STEM Plate camera for glass or film plates (3.25 x 4†plates) with 60 plate carriers and two sets of sender/receiver boxes 35-mm camera (for mounting on side port) STEM detectors Secondary Back-scattered Bright-field Annular dark-field
Specimen holders Standard two-grid holder EM-STH10: two-axis rotating holder EM-SCSH10: low-background holder EM-SRH10: specimen tilt-rotation holder EM-SCH: cryogenic holder and controller Gatan heating holder and controller
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
==============================Original Headers============================== 8, 20 -- From gary.m.brown-at-exxonmobil.com Tue Oct 17 14:44:57 2006 8, 20 -- Received: from hoespc01.exxonmobil.com (hoespc01.exxonmobil.com [192.67.48.38]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HJivTp023383 8, 20 -- for {microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:44:57 -0500 8, 20 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 8, 20 -- by hoespc01.exxonmobil.com (Switch-3.1.10/Switch-3.1.7) with ESMTP id k9HJiutp002521 8, 20 -- for {microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:44:56 -0500 (CDT) 8, 20 -- Subject: Disposal of JEOL 2000FX TEM 8, 20 -- Importance: 8, 20 -- To: microscopy-at-microscopy.com 8, 20 -- X-Mailer: Lotus Notes 652HF109 January 14, 2005 8, 20 -- Message-ID: {OFBA38CA8C.4AEE5F37-ON8625720A.006BCD58-8625720A.006C7B13-at-exxonmobil.com} 8, 20 -- From: gary.m.brown-at-exxonmobil.com 8, 20 -- Date: Tue, 17 Oct 2006 14:44:52 -0500 8, 20 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652FP1HF193|March 8, 20 -- 02, 2006) at 10/17/2006 02:44:55 PM 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-type: text/plain; charset=UTF-8 8, 20 -- Content-Transfer-Encoding: 8bit 8, 20 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k9HJivTp023383 ==============================End of - Headers==============================
1) low energy secondary electrons collection will require an additional electrode at some +300V, same as standard SED has;
2) poor vacuum of specimen chamber will likely poison high resistance layer of the channel electron multiplier. These require much better vacuum.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com ----- Original Message ----- X-from: {gary-at-gaugler.com} To: {vitalylazar-at-att.net} Sent: Monday, October 16, 2006 8:18 PM
==============================Original Headers============================== 7, 23 -- From vitalylazar-at-att.net Tue Oct 17 14:50:19 2006 7, 23 -- Received: from mtiwmhc11.worldnet.att.net (mtiwmhc11.worldnet.att.net [204.127.131.115]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HJoJkJ003771 7, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:50:19 -0500 7, 23 -- Received: from siap6segs20pa8 (burnham.ljcrf.edu[192.231.106.2]) 7, 23 -- by worldnet.att.net (mtiwmhc11) with SMTP 7, 23 -- id {20061017195018111009m86ue} ; Tue, 17 Oct 2006 19:50:18 +0000 7, 23 -- Message-ID: {005b01c6f225$755711b0$0cf910ac-at-siap6segs20pa8} 7, 23 -- From: "Vitaly Feingold" {vitalylazar-at-att.net} 7, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 7, 23 -- References: {200610170018.k9H0IDkr026649-at-ns.microscopy.com} 7, 23 -- Subject: Re: [Microscopy] Re: PMT 7, 23 -- Date: Tue, 17 Oct 2006 15:50:07 -0400 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-Type: text/plain; 7, 23 -- format=flowed; 7, 23 -- charset="iso-8859-1"; 7, 23 -- reply-type=original 7, 23 -- Content-Transfer-Encoding: 7bit 7, 23 -- X-Priority: 3 7, 23 -- X-MSMail-Priority: Normal 7, 23 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 7, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
Actually you may simply skip this step. You can do the infiltration steps with a mixture of resin and 100% ethanol.
Regards,
Stephane
----- Original Message ---- X-from: "robert.clark-at-sharp.com" {robert.clark-at-sharp.com} To: nizets2-at-yahoo.com Sent: Tuesday, October 17, 2006 1:51:48 AM
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Email: robert.clark-at-sharp.com Name: Rob
Organization: Sharp HealthCare
Title-Subject: [Filtered] TEM-Propylene Oxide v. Acetone
Question: Hi, What are the opions on using propylene oxide v. acetone mixed with resin in the final steps of resin infiltration of TEM processing? We use P.O. and have for years. The question came up as I am processing a piece of rubber tubing and am concerned the P.O. may eat away at it?
I would be careful about this - I have found that traces of alcohol can adversely affect polymerisation. I do have to use ethanol/resin mix when processing cells that have been cultured on plastic dishes, but always give extra changes in pure resin in a vacuum incubator to ensure removal of trace ethanol.
For info - it is our normal practice to use acetone as a link reagent, having changed from using PO some years ago.
Alastair
At 04:12 18/10/2006 -0500, you wrote:
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Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 11, 18 -- From a.d.mckinnon-at-abdn.ac.uk Wed Oct 18 05:07:14 2006 11, 18 -- Received: from mailhub1.abdn.ac.uk (mailhub1.abdn.ac.uk [139.133.7.28]) 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IA7DgV015183 11, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 05:07:13 -0500 11, 18 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 11, 18 -- by mailhub1.abdn.ac.uk with esmtp (Exim 4.52) 11, 18 -- id 1Ga8KO-0003JN-En; Wed, 18 Oct 2006 11:07:12 +0100 11, 18 -- Message-Id: {5.2.1.1.0.20061018105834.01137098-at-mailms.abdn.ac.uk} 11, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 11, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 11, 18 -- Date: Wed, 18 Oct 2006 11:07:10 +0100 11, 18 -- To: nizets2-at-yahoo.com 11, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 11, 18 -- Subject: Re: [Microscopy] Re: viaWWW: TEM-Propylene Oxide v. Acetone 11, 18 -- Cc: Microscopy-at-microscopy.com 11, 18 -- In-Reply-To: {200610180912.k9I9CHJa012381-at-ns.microscopy.com} 11, 18 -- Mime-Version: 1.0 11, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================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 hx213-at-cam.ac.uk as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: hx213-at-cam.ac.uk Name: Huixin Xiu
Organization: University of Cambridge
Title-Subject: [Filtered] About the defect contrast in HAADF image
Question: Dear Microscopists,
I observed a strong defect ( possibly inversion domains) contrast in STEM-HAADF image at edge-on condition in GaN. The contrast also exists along {11-20} zone axis, but it is weaker than edge-on condition. I tried to use EDX to see whether any chemical change across the defect, but didn't find obvious change. Could anybody have similar observations or have any idea of the contrast from? Thanks a lot.
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Email: rfklie-at-uic.edu Name: Robert Klie
Organization: University of Illinois - Chicago
Title-Subject: [Filtered] Postdoctoral Position available
Question: Dear all,
I am currently looking for a postdoctoral research associate in the area of STEM/EELS of oxide materials at the University of Illinois - Chicago. Would you please bring the advertisement (see below) to the attention of possible candidates?
Thanks!
Robert Klie -----------------------------------
POSTDOCTORAL POSITION AVAILABLE STARTING JANUARY 2007 DEPARTMENT OF PHYSICS UNIVERSITY OF ILLINOIS - CHICAGO, IL, USA
The University of Illinois ñ Chicago (UIC), Department of Physics, is seeking candidates for a postdoctoral position in transmission electron microscopy of strongly correlated oxides and heterogeneous catalysts systems. Applicants should have a Ph.D. degree in Materials Physics or Materials Chemistry, extensive and demonstrated experience in Z-contrast imaging in combination with electron energy-loss spectroscopy (EELS), and a strong background and interest in materials problem solving. Familiarity with aberration correction and analysis of structural defects and interfaces using electron-microscopy imaging is an advantage.
Facilities at UIC include a JEOL 2010F STEM/TEM, an aberration-corrected VG HB601UX, both with HAADF detectors and EELS spectrometers, a JEOL3010, as well as state-of-the-art sample preparation (for more details see http://www.rrc.uic.edu/).
The position is available starting January, 2007 for a period one year with a possible extension for a second year. The salary is commensurate with experience. Interested candidates should send a curriculum vitae, publication list, and the names of at least three references with their contact addresses to: Professor Robert F. Klie at rfklie-at-uic.edu
-------------------------------------------------------------------------------- Robert F. Klie, PhD Center for Functional Nanomaterials (Bldg.480) Brookhaven National Laboratory Upton, NY 11973 Tel. (631) 344-7709 Fax. (631)344-4071
AND
Assistant Professor University of Illinois at Chicago Department of Physics Chicago, IL 60607
There is a postdoctoral vacancy at the Universidad Técnica Federico Santa María. This is in the Vina del Mar/Valparaiso region of Chile. It is a good university and a great place to live. I recommend it for your consideration. Details below.
Alwyn Eades
Universidad Técnica Federico Santa María Condensed Mater Physics Group
3-years Postdoc position on Nanoscale Materials
Programa Bicentenario de Ciencia y Tecnología.
Our Condensed Mater Physics group at USM is focusing its research in both theoretical and experimental nanoscale materials. The current topics of research are: CVD synthesis of carbon nanotubes and other carbon nanostructures, synthesis of nanoparticle-nanotube hybrids, STM- AFM studies on the growth of metal thin films on crystalline surfaces in systems such as Ag/Al(100), Fe/CuAl(100), Fe/ZnO(100); Inverse Photoemission Spectroscopy of thin films and nanostructures; Low energy ion scattering studies of nanostructured materials; Theoretical studies of electronic structure of nanostructures under external electromagnetic fields.
We have one post-doc position open for somebody who fits with our research motivations. We are looking for a dynamic individual with expertise in either theory or experimental areas.
Essential requirements for candidates are: 1) A PhD in experimental or theoretical Physics, Chemistry or Materials Science. 2) A track record of quality publications 3) For experimentalists, experience with UHV and electron spectroscopies is desirable but not necessary. 4) For theorists, previous work on nanoscale systems and materials is required. 5) Excellent writing and communication skills are desired.
The position is initially opened for one year, and available immediately. Renewal up to 3-years upon mutual agreement.
Please send your CV and list of publications to Dr. Patricio Häberle (patricio.haberle-at-usm.cl)
-- ........... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
==============================Original Headers============================== 13, 24 -- From jae5-at-lehigh.edu Wed Oct 18 09:30:09 2006 13, 24 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IEU97O019280 13, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 09:30:09 -0500 13, 24 -- Received: from [127.0.0.1] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 13, 24 -- (authenticated bits=0) 13, 24 -- by rain.CC.Lehigh.EDU (8.13.8/8.13.8) with ESMTP id k9IEU73K028265 13, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 13, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 10:30:07 -0400 13, 24 -- Message-ID: {45363A6F.8000904-at-lehigh.edu} 13, 24 -- Date: Wed, 18 Oct 2006 10:30:07 -0400 13, 24 -- From: Alwyn Eades {jae5-at-lehigh.edu} 13, 24 -- Organization: Lehigh University 13, 24 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 13, 24 -- MIME-Version: 1.0 13, 24 -- To: MSA listserver {Microscopy-at-MSA.Microscopy.Com} 13, 24 -- Subject: Postdoctoral vacancy 13, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-Virus-Scanned: ClamAV version 0.88.5, clamav-milter version 0.88.5 on rain.CC.Lehigh.EDU 13, 24 -- X-Virus-Status: Clean 13, 24 -- X-Spam-Status: No, score=-1.4 required=5.0 tests=ALL_TRUSTED 13, 24 -- autolearn=disabled version=3.1.7 13, 24 -- X-Spam-Checker-Version: SpamAssassin 3.1.7 (2006-10-05) on rain.CC.Lehigh.EDU ==============================End of - Headers==============================
Today, I shot some images of histological sections in a conventional light microscope (Leica Orthoplan 2) using several different digital camera systems (Pixera, Qimaging). None of the images were uniformly in focus (i.e., they were not flat field) even though the microscope image itself was.
Is this a common problem with digital camera optical couplers? If anyone has a system that would work with this microscope setup, I would like to hear about it.
Frustrated in Carbondale,
J Bozzola -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
==============================Original Headers============================== 4, 18 -- From bozzola-at-siu.edu Wed Oct 18 15:40:25 2006 4, 18 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKeL3d004713 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 15:40:23 -0500 4, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 4, 18 -- by abbmta1.siu.edu (Switch-3.1.10/Switch-3.1.10) with ESMTP id k9IKeFuL021132 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 15:40:17 -0500 (CDT) 4, 18 -- Mime-Version: 1.0 4, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 4, 18 -- Message-Id: {p06110427c15c3dccf719-at-[131.230.177.142]} 4, 18 -- Date: Wed, 18 Oct 2006 15:40:27 -0500 4, 18 -- To: Microscopy-at-msa.microscopy.com 4, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 4, 18 -- Subject: LM: digital camera lacking flat field 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 18 -- X-Spam-Score: 0.00% 4, 18 -- X-MASF: 0.00% 4, 18 -- X-Whitelist: 0.00% ==============================End of - Headers==============================
The sharpness of EBSD patterns decreases as the SEM kV is reduced.
Does anybody know why?
The only explanation I can come up with is stray fields. The lower kV electrons are going to be more affected by stray fields.
Regards, -- Larry Stoter
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==============================Original Headers============================== 5, 14 -- From larry-at-cymru.freewire.co.uk Wed Oct 18 15:46:12 2006 5, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKkBWi009630 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 15:46:12 -0500 5, 14 -- Received: from [217.154.251.81] (th6dc-217-154-251-81.dial.mistral.co.uk [217.154.251.81] (may be forged)) 5, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9IKk9DI027032 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 21:46:10 +0100 5, 14 -- Mime-Version: 1.0 5, 14 -- Message-Id: {p06210200c15c4186a7b4-at-[217.154.250.196]} 5, 14 -- Date: Wed, 18 Oct 2006 21:43:02 +0100 5, 14 -- To: Microscopy-at-MSA.Microscopy.Com 5, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 14 -- Subject: EBSD - kV dependence of pattern sharpness 5, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Dear Larry, I would suspect that either you are getting more contribution from the amorphous surface oxide layer as the beam penetrates less at lower kV, or that you are getting more contribution from the thin, deformed layer left over from polishing. The EBSP electrons come from very shallow in the specimen, because the ones from deeper in the sample don't come out with enough energy to contribute to the pattern, so the pattern is very sensitive to the condition of the surface and top micron of the sample. IMHO. Regards,
Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca -----Original Message----- X-from: larry-at-cymru.freewire.co.uk [mailto:larry-at-cymru.freewire.co.uk] Sent: Wednesday, October 18, 2006 1:51 PM To: mager-at-interchange.ubc.ca
The sharpness of EBSD patterns decreases as the SEM kV is reduced.
Does anybody know why?
The only explanation I can come up with is stray fields. The lower kV electrons are going to be more affected by stray fields.
Regards, -- Larry Stoter
PLEASE NOTE - Agressive SPAM filtering is employed. If you don't get a reply to e-mails, try again, avoiding anything in the subject or body which might trigger filtering. If you are in the address book, you should get through. If you aren't, then there's a chance your e-mail will never be seen.
==============================Original Headers============================== 5, 14 -- From larry-at-cymru.freewire.co.uk Wed Oct 18 15:46:12 2006 5, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKkBWi009630 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 15:46:12 -0500 5, 14 -- Received: from [217.154.251.81] (th6dc-217-154-251-81.dial.mistral.co.uk [217.154.251.81] (may be forged)) 5, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9IKk9DI027032 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 21:46:10 +0100 5, 14 -- Mime-Version: 1.0 5, 14 -- Message-Id: {p06210200c15c4186a7b4-at-[217.154.250.196]} 5, 14 -- Date: Wed, 18 Oct 2006 21:43:02 +0100 5, 14 -- To: Microscopy-at-MSA.Microscopy.Com 5, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 14 -- Subject: EBSD - kV dependence of pattern sharpness 5, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 13, 33 -- From mager-at-interchange.ubc.ca Wed Oct 18 16:19:11 2006 13, 33 -- Received: from mta6.mail-relay.ubc.ca (mta6.mail-relay.ubc.ca [137.82.45.12]) 13, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ILJBre026521 13, 33 -- for {microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 16:19:11 -0500 13, 33 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 13, 33 -- by mta6.mail-relay.ubc.ca (8.12.11.20060308/8.12.11) with ESMTP id k9ILJ4CV024400 13, 33 -- for {microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 14:19:04 -0700 (PDT) 13, 33 -- (envelope-from mager-at-interchange.ubc.ca) 13, 33 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 13, 33 -- by smtp.interchange.ubc.ca 13, 33 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 13, 33 -- with ESMTPA id {0J7C00JM8OJSC4-at-smtp.interchange.ubc.ca} for 13, 33 -- microscopy-at-microscopy.com; Wed, 18 Oct 2006 14:19:04 -0700 (PDT) 13, 33 -- Date: Wed, 18 Oct 2006 14:19:34 -0700 13, 33 -- From: Mary Mager {mager-at-interchange.ubc.ca} 13, 33 -- Subject: RE: [Microscopy] EBSD - kV dependence of pattern sharpness 13, 33 -- In-reply-to: {200610182050.k9IKokZ5023599-at-ns.microscopy.com} 13, 33 -- To: larry-at-cymru.freewire.co.uk 13, 33 -- Cc: Microscopy {microscopy-at-microscopy.com} 13, 33 -- Reply-to: mager-at-interchange.ubc.ca 13, 33 -- Message-id: {0J7C00JM9OJSC4-at-smtp.interchange.ubc.ca} 13, 33 -- Organization: Materials Eng. UBC 13, 33 -- MIME-version: 1.0 13, 33 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1478 13, 33 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 13, 33 -- Content-type: text/plain; charset=us-ascii 13, 33 -- Content-transfer-encoding: 7bit 13, 33 -- Thread-index: Acby9xbMWkhIiYRETgmPcFnvh3tPMQAAkPvg 13, 33 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.18.135442 13, 33 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 13, 33 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 13, 33 -- X-Spam-Level: 13, 33 -- X-Spam-Flag: No ==============================End of - Headers==============================
Need some info on the fluid used in the water chilled lens cooling loop of a Jeol 733.
System developed a leak and dumped what at first looked like pump oil on the floor of the lab. After some clean up the fluid evaporated over the next few days which led me to think it was some kind of water based liquid.
Any 733 owners with info please contact me directly.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 9, 23 -- From rbeavers-at-mail.smu.edu Wed Oct 18 16:41:20 2006 9, 23 -- Received: from s31xub.systems.smu.edu (s31xub.systems.smu.edu [129.119.70.73]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ILfJGL004997 9, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xub.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="us-ascii" 9, 23 -- Subject: Jeol 733 Microprobe 9, 23 -- Date: Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB235E4A-at-s31xe7.systems.smu.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Jeol 733 Microprobe 9, 23 -- Thread-Index: Acby/iWDxPLNpHquTV69e0YFSFCLwA== 9, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 9, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 18 Oct 2006 21:41:19.0350 (UTC) FILETIME=[25A01160:01C6F2FE] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9ILfJGL004997 ==============================End of - Headers==============================
Roy, I've not done a whole lot of work on the JXA-733, but the objective lens is definitely OIL cooled. Don't go putting water in there! I'm just not sure what kind of oil. Maybe someone out there can help out with this.
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: rbeavers-at-mail.smu.edu [mailto:rbeavers-at-mail.smu.edu] Sent: Wednesday, October 18, 2006 5:44 PM To: kenconverse-at-qualityimages.biz
Group,
Need some info on the fluid used in the water chilled lens cooling loop of a Jeol 733.
System developed a leak and dumped what at first looked like pump oil on the floor of the lab. After some clean up the fluid evaporated over the next few days which led me to think it was some kind of water based liquid.
Any 733 owners with info please contact me directly.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 9, 23 -- From rbeavers-at-mail.smu.edu Wed Oct 18 16:41:20 2006 9, 23 -- Received: from s31xub.systems.smu.edu (s31xub.systems.smu.edu [129.119.70.73]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ILfJGL004997 9, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xub.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="us-ascii" 9, 23 -- Subject: Jeol 733 Microprobe 9, 23 -- Date: Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB235E4A-at-s31xe7.systems.smu.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Jeol 733 Microprobe 9, 23 -- Thread-Index: Acby/iWDxPLNpHquTV69e0YFSFCLwA== 9, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 9, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 18 Oct 2006 21:41:19.0350 (UTC) FILETIME=[25A01160:01C6F2FE] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9ILfJGL004997 ==============================End of - Headers==============================
==============================Original Headers============================== 22, 24 -- From kenconverse-at-qualityimages.biz Wed Oct 18 18:33:22 2006 22, 24 -- Received: from mta10.adelphia.net (mta10.adelphia.net [68.168.78.202]) 22, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9INXMNY017190 22, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 18:33:22 -0500 22, 24 -- Received: from Ken ([24.53.5.33]) by mta10.adelphia.net 22, 24 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with ESMTP 22, 24 -- id {20061018233318.LZLK14484.mta10.adelphia.net-at-Ken} ; 22, 24 -- Wed, 18 Oct 2006 19:33:18 -0400 22, 24 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 22, 24 -- To: {rbeavers-at-mail.smu.edu} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 22, 24 -- Subject: RE: [Microscopy] Jeol 733 Microprobe 22, 24 -- Date: Wed, 18 Oct 2006 19:33:14 -0400 22, 24 -- Message-ID: {002501c6f30d$c88ebd40$6401a8c0-at-Ken} 22, 24 -- MIME-Version: 1.0 22, 24 -- Content-Type: text/plain; 22, 24 -- charset="us-ascii" 22, 24 -- X-Priority: 3 (Normal) 22, 24 -- X-MSMail-Priority: Normal 22, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 22, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 22, 24 -- Importance: Normal 22, 24 -- In-Reply-To: {200610182144.k9ILiHfh009872-at-ns.microscopy.com} 22, 24 -- Content-Transfer-Encoding: 8bit 22, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9INXMNY017190 ==============================End of - Headers==============================
We are using two different couplers made by OPTEM that were designed for the specific digital camera. The couplers (containing lenses) were not inexpensive. For ports, this is the standard trinocular (camera) port on top of the oculars. Previously, it was used with 35 mm cameras and worked beautifully with film....... sigh.....
JB
} I've coupled my digital cameras to several optical microscopes in my } lab without the problem you have expressed. It sounds to me like } whatever port you are using to couple the digital camera to the } microscope has a problem. May I ask how you made the coupling? What } port are you using? What magnification are you working at? } } I actually have a coupler I use in the field when going to outside } labs that I can use to couple my digital camera to their microscopes } if the need arises. I've only had the kind of problem you are } describing when faced with a port that has some kind of problem ... } I will say at high mags, like 1000X and higher, I have run into } problems, but I have felt it is due to the pixel resolution of my } camera.... } } dj } } On Wed, 18 Oct 2006, bozzola-at-siu.edu wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
==============================Original Headers============================== 5, 21 -- From bozzola-at-siu.edu Wed Oct 18 18:39:41 2006 5, 21 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9INddWd001310 5, 21 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 18:39:41 -0500 5, 21 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 5, 21 -- by abbmta1.siu.edu (Switch-3.1.10/Switch-3.1.10) with ESMTP id k9INdcFR026474 5, 21 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 18:39:39 -0500 (CDT) 5, 21 -- Mime-Version: 1.0 5, 21 -- X-Sender: bozzola-at-saluki-mail.siu.edu 5, 21 -- Message-Id: {p0611042ac15c6b219726-at-[131.230.177.142]} 5, 21 -- In-Reply-To: {Pine.WNT.4.64.0610181702210.3148-at-H-F1} 5, 21 -- References: {200610182047.k9IKloI7015067-at-ns.microscopy.com} 5, 21 -- {Pine.WNT.4.64.0610181702210.3148-at-H-F1} 5, 21 -- Date: Wed, 18 Oct 2006 18:39:49 -0500 5, 21 -- To: Microscopy-at-msa.microscopy.com 5, 21 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 5, 21 -- Subject: Re: [Microscopy] LM: digital camera lacking flat field 5, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 21 -- X-Spam-Score: 0.00% 5, 21 -- X-MASF: 0.00% 5, 21 -- X-Whitelist: 0.00% ==============================End of - Headers==============================
Just an update on our faulty Hitachi H-600 CRT. Our electrician, Tony Morgan, has fixed the problem. We didn't have 15V entering the CRT unit. He traced the fault back to circuit board PC-1 in the power box. The volatage regultor for fuse F6 had a "dry joint". He removed PC-1 and resoldered the pins on the voltage regulator and the fuse clips. He also replaced the fuse.
This would explain why the CRT only lasted 10 minutes each morning - as the power box heated, the slightest amount of expansion in the "dry joint" was causing the loss of continuity in the circuit.
Thanks to everyone who provided advice on this issue.
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia
==============================Original Headers============================== 6, 33 -- From john.brealey-at-imvs.sa.gov.au Wed Oct 18 19:01:27 2006 6, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9J01QmC016733 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 19:01:26 -0500 6, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 6, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9J01OJZ001259 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:31:24 +0930 (CST)' 6, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 6, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9J01O3Y001256 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:31:24 +0930 (CST)' 6, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 6, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id BC98134D3E 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:31:24 +0930 (CST) 6, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 6, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 6, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 6, 33 -- with LMTP id YjK3ws2u5hLc for {Microscopy-at-MSA.Microscopy.Com} ; 6, 33 -- Thu, 19 Oct 2006 09:31:16 +0930 (CST) 6, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 6, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 8212634D63 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:30:32 +0930 (CST) 6, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 6, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 6, 33 -- Subject: Hitachi H-600 CRT Fixed 6, 33 -- Date: Thu, 19 Oct 2006 09:30:30 +0930 6, 33 -- Message-ID: {000001c6f311$9822f820$c88a140a-at-iqe36042} 6, 33 -- MIME-Version: 1.0 6, 33 -- Content-Type: text/plain; 6, 33 -- charset="us-ascii" 6, 33 -- Content-Transfer-Encoding: 7bit 6, 33 -- X-Mailer: Microsoft Office Outlook 11 6, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 6, 33 -- Thread-Index: AcbzEXNlC41UFSJlSR22Qp7NrxzA1g== ==============================End of - Headers==============================
Isabell and Dravid have considered the problem in a paper on "Resolution and sensitivity of electron backscattered diffraction in a cold field emission gun SEM" in Ultramicroscopy 67 (1997) 59-68.
Concerning the stray fields that you mention as a possible explanation they write: "Some high-resolution SEMs have a strong objective lens field which "spills" onto the specimen. This results in bending of Kikuchi lines due to the excessive magnetic field of the objective lens. This problem, however, can be easily solved by placing a small aperture of permalloy above the specimen, thus trapping the magnetic flux from the lens."
More generally, they conclude that you get a lousy signal-to-noise ratio at low voltages because low energy backscattered electrons are much less efficient in exciting the phosphor than high energy BSEs. Hence the pattern quality is degraded at low voltages.
-----Original Message----- X-from: larry-at-cymru.freewire.co.uk [mailto:larry-at-cymru.freewire.co.uk] Sent: Wednesday, October 18, 2006 10:51 PM To: j.bilde-at-risoe.dk
The sharpness of EBSD patterns decreases as the SEM kV is reduced.
Does anybody know why?
The only explanation I can come up with is stray fields. The lower kV electrons are going to be more affected by stray fields.
Regards, -- Larry Stoter
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==============================Original Headers============================== 5, 14 -- From larry-at-cymru.freewire.co.uk Wed Oct 18 15:46:12 2006 5, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKkBWi009630 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 15:46:12 -0500 5, 14 -- Received: from [217.154.251.81] (th6dc-217-154-251-81.dial.mistral.co.uk [217.154.251.81] (may be forged)) 5, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9IKk9DI027032 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 21:46:10 +0100 5, 14 -- Mime-Version: 1.0 5, 14 -- Message-Id: {p06210200c15c4186a7b4-at-[217.154.250.196]} 5, 14 -- Date: Wed, 18 Oct 2006 21:43:02 +0100 5, 14 -- To: Microscopy-at-MSA.Microscopy.Com 5, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 14 -- Subject: EBSD - kV dependence of pattern sharpness 5, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 20, 25 -- From j.bilde-at-risoe.dk Thu Oct 19 04:27:50 2006 20, 25 -- Received: from mail.risoe.dk (mail.risoe.dk [130.226.48.21]) 20, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9J9RnNC004833 20, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 04:27:50 -0500 20, 25 -- Received: from EXCHG-VS1.risoe.dk (exchg-01.risoe.dk [10.0.0.26]) 20, 25 -- by risoe.dk (PMDF V6.2-X26 #31238) with ESMTP id {IKYU08R1H6CS84029C-at-risoe.dk} 20, 25 -- for Microscopy-at-microscopy.com; Thu, 20, 25 -- 19 Oct 2006 11:27:34 +0200 (Romance Daylight Time) 20, 25 -- Date: Thu, 19 Oct 2006 11:27:33 +0200 20, 25 -- From: j.bilde-at-risoe.dk 20, 25 -- Subject: RE: [Microscopy] EBSD - kV dependence of pattern sharpness 20, 25 -- In-reply-to: {200610182051.k9IKpMLa025119-at-ns.microscopy.com} 20, 25 -- To: larry-at-cymru.freewire.co.uk 20, 25 -- Cc: Microscopy-at-microscopy.com 20, 25 -- Message-id: {6463F256A85DC14CAB07F087A2479972AFEA69-at-EXCHG-VS1.risoe.dk} 20, 25 -- MIME-version: 1.0 20, 25 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 20, 25 -- Content-type: text/plain; charset=iso-8859-1 20, 25 -- Thread-Topic: [Microscopy] EBSD - kV dependence of pattern sharpness 20, 25 -- Thread-Index: Acby9z96W3zLxO4HTHKxG6OpMnJ+1wAZ6fJg 20, 25 -- Content-class: urn:content-classes:message 20, 25 -- X-MS-Has-Attach: 20, 25 -- X-MS-TNEF-Correlator: 20, 25 -- Content-Transfer-Encoding: 8bit 20, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9J9RnNC004833 ==============================End of - Headers==============================
Applications are invited for this position in our Materials Research Group (MRG). Responsibilities will include: - Adapting of cryo-stage coupled to the MRG nuclear microprobe for measurements of thin specimens in frozen-hydrated state - Active involvement in research projects related to biological applications of ion beam techniques and generating new applications.
Minimum requirements:
- PhD in Biology/Chemistry/Physics with strong emphasis on cryo-preparation of biological specimens and low temperature electron microscopy - Experience in operating SEM/TEM and knowledge of EDS technique as well as cryo-ultramicrotomy - Experience in operating a nuclear microprobe and familiarity with PIXE as an advantage - Relevant conference presentations and publications as well as international exposure - Knowledge of statistical methods - Computer literacy (Word, Excel, Corel, etc.)
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Applications close on 3 November 2006
Please contact me directly for any more information on this position
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa E-mail: przybylowicz-at-tlabs.ac.za Fax: +27-21-8433543 Phone: +27-21-8431166 (direct); 8431000 (reception) Cell: +27-82-563 7925 http://www.tlabs.ac.za xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx P
==============================Original Headers============================== 10, 29 -- From przybylowicz-at-tlabs.ac.za Thu Oct 19 06:11:14 2006 10, 29 -- Received: from silence.tlabs.ac.za (silence.tlabs.ac.za [196.21.124.19]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9JBBB95016949 10, 29 -- for {microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 06:11:12 -0500 10, 29 -- Received: from segonyane.tlabs.ac.za ([196.21.125.83]:3132 helo=przybylo) 10, 29 -- by silence.tlabs.ac.za with smtp (Exim 4.43) 10, 29 -- id 1GaVnZ-0002dw-JE 10, 29 -- for microscopy-at-microscopy.com; Thu, 19 Oct 2006 13:10:53 +0200 10, 29 -- Message-ID: {036d01c6f36f$0bb57a80$537d15c4-at-przybylo} 10, 29 -- From: "Przybylowicz Wojciech" {przybylowicz-at-tlabs.ac.za} 10, 29 -- To: {microscopy-at-microscopy.com} 10, 29 -- Subject: Post-doctoral fellow: Nuclear microprobe of thin frozen-hydrated biological specimens 10, 29 -- Date: Thu, 19 Oct 2006 13:09:23 +0200 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain; 10, 29 -- charset="iso-8859-1" 10, 29 -- Content-Transfer-Encoding: 7bit 10, 29 -- X-Priority: 3 10, 29 -- X-MSMail-Priority: Normal 10, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1106 10, 29 -- Disposition-Notification-To: "Przybylowicz Wojciech" {przybylowicz-at-tlabs.ac.za} 10, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1106 10, 29 -- X-tlabs-Information: Please contact iThemba LABS, South Africa 10, 29 -- X-tlabs-MailScanner: Found to be clean 10, 29 -- X-tlabs-SpamCheck: not spam (whitelisted), SpamAssassin (not cached, 10, 29 -- score=-4.568, required 6, autolearn=not spam, ALL_TRUSTED -3.30, 10, 29 -- BAYES_00 -2.60, CELL_PHONE_IMPROVE 1.03, SARE_WEOFFER 0.30) 10, 29 -- X-tlabs-MailScanner-From: przybylowicz-at-tlabs.ac.za 10, 29 -- X-tlabs-MailScanner-To: microscopy-at-microscopy.com ==============================End of - Headers==============================
Hi, I'm not sure what JEOL uses on the JXA 733, but on the JXA 8600 they use an oil in that cooling circuit. Is this in part due to the fact that on the 8600 the voltage lines to the lens run inside the cooling lines? Was the 733 designed the same way? If it is the same, JEOL has the oil as:
Catalog Number Description Unit Price 423003 Lens cooling oil (per gallon) $13.96
Mike
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==============================Original Headers============================== 11, 12 -- From mmcheath-at-mailbox.syr.edu Thu Oct 19 06:16:42 2006 11, 12 -- Received: from mailer.syr.edu (mailer.syr.edu [128.230.18.29]) 11, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9JBGgVY026947 11, 12 -- for {microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 06:16:42 -0500 11, 12 -- Received: from [128.230.24.90] (www.geochemistry.syr.edu) by mailer.syr.edu (LSMTP for Windows NT v1.1b) with SMTP id {0.15978AAD-at-mailer.syr.edu} ; Thu, 19 Oct 2006 7:16:41 -0400 11, 12 -- Mime-Version: 1.0 11, 12 -- Message-Id: {a06230907c15d0e5934de-at-[128.230.24.90]} 11, 12 -- Date: Thu, 19 Oct 2006 07:16:38 -0400 11, 12 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 11, 12 -- From: Michael Cheatham {mmcheath-at-mailbox.syr.edu} 11, 12 -- Subject: Fwd: [Microscopy] RE: Je