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Question: We are looking for manuals for an old controller and photomultiplier tube for a fluorescence spectroscopy system. The set up was used in the mid 90s and we are hoping to use it again but cannot find the manuals for these components.
The controller is an Action Research Corporation model number 275. The photomultiplier tube is from Research Inc. in Davers, MA model number R636/0115/0381.
Please let me know if anyone has copies of these manuals and we could pay for copying and shiping.
I used clean and subbed CR-39 plastic or glass 3x1" dlides and Spurr's resin embedding (or occasionally LR White I think). The Spurr's resin was removed by soaking the slides for around 20 minutes in sodium ethoxide (1.5 NaOH in 100cm3 ethanol and aged for 1 day prior to use). Always use freshly made sodium ethoxide (sodium chloride dissolved in ethanol). Never buy the ready made stuff and always make it up fresh after a few days use - although our less descerning histology lab kept there's a few weeks when batching slides through (and lost more sections). I very rarely, if ever lost the tissue section, but I always kept an eye on the slides as they were soaking (the resin starts to sag on the slide surface). Looking at them often helps agitate the solution (although with lung it's diffiicult to see if the section has fallen off until after washing - I can't remember what I washed the slides in but I assume it was neat ethanol, and possibly steps though serial rehydration (I didn't publish that bit - it must be in my lab notebooks in the attic). I used a standard (at the time) carousel plastic slide holder that could hold 50 slides in a large'ish beaker (Just Plastics plc). Our workshop made a PTFE ring to hold the slide carousel down as it had a tendency to float with a few glass or all CR-39 slides. CR-39 slides were used to create 235-UO2 fission frgament autoradiographs and glass slides (the serial section) were stained with H&E. This method doesn't 'etch' Spurr resin - I assume you want to completely remove from the section. We never had any residual Spurr's on the slide (it would have interfered with our 'soft alpha particle' shadowing technique used to visualise the tissue section into the CR-39 plastic and made us cross) .
Our (mainly lung) tissue was always fixed in freon FC-80 dissolved 1% Osmium tetroxide, followed by fixation in 3% gluteraldehyde storage in 0.1M sodium dicodylate buffer (4oC). Although fixing to EM standards for section quality, we viewed under the light microscope with a Magiscan Colour image analyser system.
Keith
------------------------------------------------ Dr Keith J Morris Cell Biology Division Institute of Ophthalmology University College London 11-43 bath Street London EC1V 9EL
Tel: 020 7608 4050
----- Original Message ----- X-from: {mayas003-at-yahoo.com} To: {keith.morris-at-ucl.ac.uk} Sent: Friday, June 30, 2006 5:31 PM
I need to replace my old critical point dryer and was wondering if anyone has any leads on the purchase of a previously owned one. Any assistance is much appreciated. Thanks.
Kelly
-- S. Kelly Sears, Ph.D., B.F.A. Facility for Electron Microscopy Research McGill University
==============================Original Headers============================== 6, 21 -- From sksears-at-eps.mcgill.ca Tue Jul 4 13:37:39 2006 6, 21 -- Received: from torrent.cc.mcgill.ca (torrent.CC.McGill.CA [132.206.27.49]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k64IbdVk015835 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Jul 2006 13:37:39 -0500 6, 21 -- Received: from mailscan5.CC.McGill.CA (mailscan5.CC.McGill.CA [132.216.77.252]) 6, 21 -- by torrent.cc.mcgill.ca (8.12.11/8.12.3) with ESMTP id k64Ibcch008649 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Jul 2006 14:37:38 -0400 6, 21 -- Received: from eps.mcgill.ca (dhcp21643226.Medicine.McGill.CA [132.216.43.226]) 6, 21 -- by mailscan5.CC.McGill.CA (8.12.11/8.12.11) with ESMTP id k64IbQdi031597 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Jul 2006 14:37:27 -0400 6, 21 -- Message-ID: {44AAB56B.803-at-eps.mcgill.ca} 6, 21 -- Date: Tue, 04 Jul 2006 14:37:31 -0400 6, 21 -- From: "S. Kelly Sears" {sksears-at-eps.mcgill.ca} 6, 21 -- Organization: Facility for Electron Microscopy Research 6, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 6, 21 -- X-Accept-Language: en-us, en 6, 21 -- MIME-Version: 1.0 6, 21 -- To: Microscopy-at-microscopy.com 6, 21 -- Subject: Needed: Critical Point Dryer 6, 21 -- Content-Type: text/plain; charset=us-ascii; format=flowed 6, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am wondering if hydrofluoric acid technique to remove glass coverslips from cultured cell monolayers is compatible with LR White resin and subsequent immunolabeling. I mostly see in the Listserv archives references to Araldite or Embed 812. Any thoughts would be appreciated.
Best Regards, Kirk
Kirk J. Czymmek, Ph.D. Associate Professor Department of Biological Sciences University of Delaware Newark, DE 19716
==============================Original Headers============================== 5, 18 -- From kirk-at-UDel.Edu Wed Jul 5 08:00:54 2006 5, 18 -- Received: from copland.udel.edu (copland.udel.edu [128.175.13.92]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65D0snr009736 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 08:00:54 -0500 5, 18 -- Received: from [128.175.253.124] (host-253-124.nss.udel.edu [128.175.253.124]) 5, 18 -- by copland.udel.edu (8.13.6/8.13.6) with ESMTP id k65D0rrJ018283 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 09:00:53 -0400 (EDT) 5, 18 -- Message-ID: {44ABBAC1.7030305-at-udel.edu} 5, 18 -- Date: Wed, 05 Jul 2006 09:12:33 -0400 5, 18 -- From: kirk czymmek {kirk-at-UDel.Edu} 5, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.8) Gecko/20050511 5, 18 -- X-Accept-Language: en-us, en 5, 18 -- MIME-Version: 1.0 5, 18 -- To: Microscopy-at-microscopy.com 5, 18 -- Subject: Hydrofluoric acid and LR White 5, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 5, 18 -- Content-Transfer-Encoding: 7bit 5, 18 -- X-Scanned-By: MIMEDefang 2.52 on 128.175.13.92 ==============================End of - Headers==============================
} I am wondering if hydrofluoric acid technique to remove glass coverslips } from cultured cell monolayers is compatible with LR White resin and } subsequent immunolabeling. I mostly see in the Listserv archives references to Araldite or Embed 812. Any thoughts would be appreciated.
Dear Kirk, I just tried that very thing recently, with less-than-stellar results. The block faces were soft and difficult to cut. I got zero labelling, but I'm not sure what that was due to. I do know that my secondary antibodies were fine (I tested them on a known sample). Now, I must say that I do not know if the blocks were soft because of the HF treatment, or because they hadn't polymerized fully in the first place...there were some trapped air bubbles that may have interfered with it. In hindsight, my advise would be to make up a few blank, dummy blocks to test it out.
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============================== 4, 23 -- From lcgould-at-med.cornell.edu Wed Jul 5 08:41:16 2006 4, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65DfGEX020361 4, 23 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 08:41:16 -0500 4, 23 -- Received: from mpx2.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 4, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k65DfCRD026497 4, 23 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 09:41:13 -0400 (EDT) 4, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 4, 23 -- by mpx2.med.cornell.edu 4, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 4, 23 -- with ESMTPA id {0J1X00K5DNCN9X10-at-mpx2.med.cornell.edu} for 4, 23 -- microscopy-at-microscopy.com; Wed, 05 Jul 2006 09:41:12 -0400 (EDT) 4, 23 -- Date: Wed, 05 Jul 2006 09:33:40 -0400 4, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 4, 23 -- Subject: Re: [Microscopy] Hydrofluoric acid and LR White 4, 23 -- In-reply-to: {200607051302.k65D26In010993-at-ns.microscopy.com} 4, 23 -- Sender: lcgould-at-med.cornell.edu 4, 23 -- To: kirk-at-UDel.Edu, Microscopy Listserver {microscopy-at-microscopy.com} 4, 23 -- Message-id: {p06230904c0d16e45d76b-at-[140.251.48.23]} 4, 23 -- MIME-version: 1.0 4, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 4, 23 -- References: {200607051302.k65D26In010993-at-ns.microscopy.com} 4, 23 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.7.5.62432 ==============================End of - Headers==============================
I just share with you a trick I got from this list itself some time ago. I tried it with success and now I use it routinely. To obtain perfectly flat surface of resin-embedded cell monolayers, you just have to:
- Prepare open capsules by cutting off the dead end of beam capsules. - Grow your cells on coverslips - When it is time to embed, use just enough resin as to cover the coverslip with a few mm (not ml) resin (it is about 1-1.5 ml for a 3.5cm petri dish). Then return a beam capsule on your coverslip, press firmly so there is no space between the capsule and the coverslip. - Cure again. During the curing, the capsule will be glued in the resin. Next day fill the capsule with fresh resin and cure again (from my own experience doubling the time of curing help detaching the blocks from the coverslips). - When the capsules are well cured, use a pair of forceps to take the capsule off the coverslip. Sometimes some of the capsule surface sticks to the coverslips but you usually always keep enough surface to be ultracut. "overcuring" really helps here. You can also use 4 capsules per coverslip, which gives you complety security (I personally use 2 capsules per coverslip and usually don't need the second). - The surface of the block is perfectly flat, the first section will be good!! (which is interesting if you have very flat cells like me)
Good luck
Stéphane
--- kirk-at-UDel.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 } ---------------------------------------------------------------------------- } } Microscopy-at-microscopy.com } } Dear Microscopists, } } I am wondering if hydrofluoric acid technique to } remove glass coverslips } from cultured cell monolayers is compatible with LR } White resin and } subsequent immunolabeling. I mostly see in the } Listserv archives } references to Araldite or Embed 812. Any thoughts } would be appreciated. } } Best Regards, Kirk } } Kirk J. Czymmek, Ph.D. } Associate Professor } Department of Biological Sciences } University of Delaware } Newark, DE 19716 } } ==============================Original } Headers============================== } 5, 18 -- From kirk-at-UDel.Edu Wed Jul 5 08:00:54 2006 } 5, 18 -- Received: from copland.udel.edu } (copland.udel.edu [128.175.13.92]) } 5, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k65D0snr009736 } 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 } Jul 2006 08:00:54 -0500 } 5, 18 -- Received: from [128.175.253.124] } (host-253-124.nss.udel.edu [128.175.253.124]) } 5, 18 -- by copland.udel.edu (8.13.6/8.13.6) with } ESMTP id k65D0rrJ018283 } 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 } Jul 2006 09:00:53 -0400 (EDT) } 5, 18 -- Message-ID: {44ABBAC1.7030305-at-udel.edu} } 5, 18 -- Date: Wed, 05 Jul 2006 09:12:33 -0400 } 5, 18 -- From: kirk czymmek {kirk-at-UDel.Edu} } 5, 18 -- User-Agent: Mozilla/5.0 (Windows; U; } Windows NT 5.0; en-US; rv:1.7.8) Gecko/20050511 } 5, 18 -- X-Accept-Language: en-us, en } 5, 18 -- MIME-Version: 1.0 } 5, 18 -- To: Microscopy-at-microscopy.com } 5, 18 -- Subject: Hydrofluoric acid and LR White } 5, 18 -- Content-Type: text/plain; charset=us-ascii; } format=flowed } 5, 18 -- Content-Transfer-Encoding: 7bit } 5, 18 -- X-Scanned-By: MIMEDefang 2.52 on } 128.175.13.92 } ==============================End of - } Headers============================== }
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I would assume that overcuring or increased embedding could well interfere with the immunolabelling.
So I can only suggest trying liquid nitrogen to free the coverslips or else using an alternative such as melinex instead of glass coverslips because that can be sectioned if necessary. I cannot speak from experience about their use in immunolabelling but have used both techniques as an alternative to HF.
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: nizets2-at-yahoo.com
Hi all
I need a software to build a panorama image, or geant image from a set of overlapping images taken by SEM.
On the web one can find dozen of freewares, sharewares or commercialy one, and I used one which works great, but they are all under windows. The only one I found running under linux is "PanTools" which should work on all OS, but I don't catch how to install and use it. It should work as a plugin for gimp, but... I tried an ImageJ plugin too, but the most part was to be done manually.
So any suggestions are welcome.
-- J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
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: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Friday, June 16, 2006 2:07 PM To: lesley.bechtold-at-jax.org
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I know Autostitch doesn't fall under the 'linux' OS, but its been *THE* best far and beyond for putting together images (in my experience).
Well worth the hassle to move to a windows platform in most cases I would guess.
*I do not have any financial/developmental interest in the software, no connection to Matthew Brown. I am just a very satisfied user of the software. Web page is www.cs.ubc.ca/~mbrown/autostitch/autostitch.html
Regards,
Geoff Williams Leduc Bioimaging Facility Manager Brown University
http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/ -----Original Message----- X-from: jacques.faerber-at-ipcms.u-strasbg.fr [mailto:jacques.faerber-at-ipcms.u-strasbg.fr] Sent: Wednesday, July 05, 2006 12:00 PM To: Williams, Geoffrey
Hi all
I need a software to build a panorama image, or geant image from a set of overlapping images taken by SEM.
On the web one can find dozen of freewares, sharewares or commercialy one, and I used one which works great, but they are all under windows. The only one I found running under linux is "PanTools" which should work on all OS, but I don't catch how to install and use it. It should work as a plugin for gimp, but... I tried an ImageJ plugin too, but the most part was to be done manually.
So any suggestions are welcome.
-- J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
On Jul 5, 2006, at 8:59 AM, jacques.faerber-at-ipcms.u-strasbg.fr wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Hi all } } I need a software to build a panorama image, or geant image from a set } of overlapping images taken by SEM. } } On the web one can find dozen of freewares, sharewares or commercialy } one, and I used one which works great, but they are all under } windows. } The only one I found running under linux is "PanTools" which should } work } on all OS, but I don't catch how to install and use it. It should work } as a plugin for gimp, but... } I tried an ImageJ plugin too, but the most part was to be done } manually. } } So any suggestions are welcome. } } } -- } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } } ==============================Original } Headers============================== } 8, 28 -- From jacques.faerber-at-ipcms.u-strasbg.fr Wed Jul 5 } 10:55:31 2006 } 8, 28 -- Received: from mailhost.u-strasbg.fr (mailhost.u- } strasbg.fr [130.79.200.157]) } 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k65FtU0M021246 } 8, 28 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 10:55:31 } -0500 } 8, 28 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr } [130.79.210.2]) } 8, 28 -- by mailhost.u-strasbg.fr (8.13.6/jtpda-5.5pre1) } with ESMTP id k65FtUTD029465 } 8, 28 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 } 17:55:30 +0200 (CEST) } 8, 28 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr } [130.79.54.3]) } 8, 28 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id B76F710000E4 } 8, 28 -- for {Microscopy-at-Microscopy.Com} ; Wed, 5 Jul 2006 } 17:55:22 +0200 (CEST) } 8, 28 -- Message-ID: {44ABE0D2.9070800-at-ipcms.u-strasbg.fr} } 8, 28 -- Date: Wed, 05 Jul 2006 17:54:58 +0200 } 8, 28 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} } 8, 28 -- User-Agent: Mozilla Thunderbird 1.0.8 (X11/20060503) } 8, 28 -- X-Accept-Language: fr, en } 8, 28 -- MIME-Version: 1.0 } 8, 28 -- To: Microscopy-at-microscopy.com } 8, 28 -- Subject: panorama software under linux } 8, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 8, 28 -- Content-Transfer-Encoding: 8bit } 8, 28 -- X-IPCMS-MailScanner: Found to be clean } 8, 28 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr } 8, 28 -- X-Greylist: Sender IP whitelisted, not delayed by milter- } greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.157]); Wed, 05 } Jul 2006 17:55:30 +0200 (CEST) } 8, 28 -- X-Virus-Scanned: ClamAV 0.88.2/1585/Tue Jul 4 22:39:34 } 2006 on mr7.u-strasbg.fr } 8, 28 -- X-Virus-Status: Clean } 8, 28 -- X-Spam-Status: No, score=-1.4 required=5.0 } tests=ALL_TRUSTED,AWL } 8, 28 -- autolearn=disabled version=3.1.1 } 8, 28 -- X-Spam-Checker-Version: SpamAssassin 3.1.1 (2006-03-10) on } mr7.u-strasbg.fr } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 23 -- From Elliott-at-arizona.edu Wed Jul 5 13:39:15 2006 9, 23 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65IdEnG031716 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 13:39:14 -0500 9, 23 -- Received: from localhost (faramir.email.arizona.edu [10.0.0.218]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id E2BF5E95932 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 11:39:02 -0700 (MST) 9, 23 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 268BBE9726A 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 11:38:51 -0700 (MST) 9, 23 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 23 -- In-Reply-To: {200607051559.k65FxFOj026814-at-ns.microscopy.com} 9, 23 -- References: {200607051559.k65FxFOj026814-at-ns.microscopy.com} 9, 23 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 23 -- Message-Id: {A38537E5-E446-4194-8DA4-37EC4DA2FBF1-at-arizona.edu} 9, 23 -- From: David Elliott {Elliott-at-arizona.edu} 9, 23 -- Subject: Re: [Microscopy] panorama software under linux 9, 23 -- Date: Wed, 5 Jul 2006 11:38:50 -0700 9, 23 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 9, 23 -- X-Mailer: Apple Mail (2.752.2) 9, 23 -- X-Virus-Scanned: amavisd-new at email.arizona.edu 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k65IdEnG031716 ==============================End of - Headers==============================
We routinely embed "Vibratome" sections between two pieces of ACLAR cut to fit on a standard 1" X 3" microscope slide (For convenience in handling--smaller or larger pieces can be used). This does not always result in a perfectly flat sample. If that is a problem then a small weight can be placed on top of the sample during polymerization. This may make a very fragile sample so just carefully remove one of the ACLAR pieces (I use a razor blade to separate the two pieces of ACLAR and then gently pull them apart) and add a few drops of epoxy to the sample and polymerize. This will give a sturdy sample that is very flat on one side. If you wish to section a small part of your sample cut out the area with a razor blade and glue (with cyanoacrylate "SuperGlue") or use epoxy resin to attach to a blank specimen block. Leave a little margin around the area of interest when cutting a sample with a razor blade because the edges may curl a bit during the cutting/gluing process.
Another quick solution if you do not have ACLAR on hand: Make some very thin epoxy sheets by spreading epoxy resin very thinly on your embedding molds. You can even use the backside of the molds to get larger pieces. Polymerization need not be complete but, for example, 24 hours of a 48 hour process. Cut out two pieces and embedd your tissue section with a couple drops of resin between them. Polymerize.
Cutting thicker sections will reduce the curling during processing but not eliminate it util you get to one millimeter or so which makes light microscopy and trimming specific areas of tissue a big problem. I have embedded many tissues, from drosophila embryos to one centimeter diameter sections of mamalian brain, with this basic technique. Best wishes Larry
dyel-at-mail.nih.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 } ---------------------------------------------------------------------------- } } 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 dyel-at-mail.nih.gov as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: dyel-at-mail.nih.gov } Name: Chip Dye } } Organization: NIH } } Title-Subject: [Filtered] Flat embedding } } Question: Dear ListServers: } } Would anyone have any recommendations on the best way to flat embed } mouse embryos (12 days old) which have been sectioned on a vibratome? } I embedded the embryos in 10% gelatin prior to sectioning and cut 50 } micron sections and embedded in Epon. It was during dehydration and } infiltration, that my sections started to curl. After polymerizing } in a flat mold, my sections looked like curled "potato chips". Would } cutting the sections thicker help with this issue? Any assistance as } to the best method to keep sections from doing this, would be greatly } appreciated. } } Regards, } } Chip Dye } } Microscopist } Microscopy & Imaging Core, NICHD, NIH } Building 49, Room 5W-14 } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } Phone: 301-496-3627 } E-mail: dyel-at-mail.nih.gov } } } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Sat Jul 1 05:34:49 2006 } 12, 12 -- Received: from [192.169.1.141] (msdvpn24.msd.anl.gov [130.202.238.88]) } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k61AYk3B004150 } 12, 12 -- for {microscopy-at-microscopy.com} ; Sat, 1 Jul 2006 05:34:48 -0500 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p06020401c0cc003743f2-at-[192.169.1.141]} } 12, 12 -- Date: Sat, 1 Jul 2006 05:34:46 -0500 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: dyel-at-mail.nih.gov (by way of MicroscopyListserver) } 12, 12 -- Subject: viaWWW: Flat embedding } 12, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
All: the URL I previous sent has no spaces in it. Must have been a finger mis-poke. http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTable/
r-holdford-at-ti.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 } ---------------------------------------------------------------------------- } } Debby and anyone else who wants it: here's a link to download the files: } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTable/ } } Debby Sherman wrote: } } } Gee..would you mind sharing that program with others? I would love to have } } a copy. } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } http://www.agriculture.purdue.edu/microscopy } } } } } } On 6/28/06 6:37 PM, "r-holdford-at-ti.com" {r-holdford-at-ti.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 } } } ---------------------------------------------------------------------------- } } } } } } Owen: one of my favorites is EM Periodic Table, a program authored by } } } Scott Walck many years ago. It has a periodic chart layout and a table } } } layout that can be translated to Excel. I got it at a Lehigh short } } } course and I find it very handy. I'd be happy to share it with you. } } } Scott calls this "beerware" meaning when we meet him at conferences, we } } } owe him a beer if we use it. } } } } } } Scott: you can claim your pitcher at M&M in Chicago. This program } } } would be cheap at twice the price. } } } } } } opmills-at-mtu.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 } } } } ---------------------------------------------------------------------------- } } } } } } } } Does anyone know where I can find a spreadsheet or text version of x- } } } } ray emission lines? } } } } } } } } Thanks, } } } } } } } } OWen } } } } } } } } } } } } } } } } } } } } Owen P. Mills } } } } Director, Materials Characterization & Fabrication Facilities } } } } Electron Optics Engineer, Applied Chemical & Morphological Analysis } } } } Laboratory } } } } } } } } Materials Science & Engineering } } } } Michigan Technological University } } } } Rm 512 M&M Bldg. } } } } Houghton, MI 49931 } } } } PH 906-369-1875 } } } } FAX 906-487-2934 } } } } mailto:opmills-at-mtu.edu } } } } http://www.mm.mtu.edu/~opmills } } } } } } } } } } } } } } } } ==============================Original Headers============================== } } } } 10, 31 -- From opmills-at-mtu.edu Wed Jun 28 14:59:39 2006 } } } } 10, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) } } } } 10, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } } } k5SJxdNZ021212 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 14:59:39 -0500 } } } } 10, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu } } } } [141.219.69.6]) } } } } 10, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id k5SJxc9N010211 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } } } } 10, 31 -- Received: from node2.edge.dcsint.mtu.edu (node2.mtu.edu } } } } [141.219.69.2]) } } } } 10, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP } } } } id k5SJxcCb016435 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } } } } 10, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) } } } } 10, 31 -- by node2.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id } } } } k5SJxbZH003257 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } } } } 10, 31 -- (envelope-from opmills-at-mtu.edu) } } } } 10, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu } } } } [141.219.69.6]) } } } } 10, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k5SJxbQP012706 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:37 -0400 } } } } (EDT) } } } } 10, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu } } } } [141.219.192.45]) } } } } 10, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc } } } } v1.0) with ESMTP id k5SJxbbF016425 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:37 -0400 } } } } 10, 31 -- Mime-Version: 1.0 (Apple Message framework v750) } } } } 10, 31 -- Content-Transfer-Encoding: 7bit } } } } 10, 31 -- Message-Id: {82FC92F2-DB6B-4300-BE2B-59C95CA02CBB-at-mtu.edu} } } } } 10, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } } } } format=flowed } } } } 10, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} } } } } 10, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} } } } } 10, 31 -- Subject: need x-ray emission table } } } } 10, 31 -- Date: Wed, 28 Jun 2006 15:59:46 -0400 } } } } 10, 31 -- X-Mailer: Apple Mail (2.750) } } } } 10, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.4.0.264935, } } } } Antispam-Data: 2006.6.28.123433 } } } } 10, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, 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' } } } } ==============================End of - Headers============================== } } } } } } } } } } } } } } } } } } } } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 22 -- From r-holdford-at-ti.com Wed Jul 5 14:50:15 2006 4, 22 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65JoDD5020806 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 14:50:15 -0500 4, 22 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 22 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k65Jo8CS019260 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 14:50:13 -0500 (CDT) 4, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 22 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k65Jo7GA024590 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 14:50:07 -0500 (CDT) 4, 22 -- Message-ID: {44AC17EF.4050803-at-ti.com} 4, 22 -- Date: Wed, 05 Jul 2006 14:50:07 -0500 4, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 22 -- Organization: SC Packaging Development -- FA Development 4, 22 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 4, 22 -- MIME-Version: 1.0 4, 22 -- To: MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Re: [Microscopy] need x-ray emission table 4, 22 -- References: {200607051814.k65IEexl011744-at-ns.microscopy.com} 4, 22 -- In-Reply-To: {200607051814.k65IEexl011744-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
The issue of flat-embedding has been addressed by several authors, including Schwartz (1982), Schafer (1989), Nguyen and Pender (1995), Larue and Winer (1996). In my processing I largely follow the approach established by Larue and Winer (1996) with some modifications (Larue DT, Winer JA. Postembedding immunocytochemistry of large sections of brain tissue: an improved flat embedding technique. J Neurosci Methods, 1996 Sep; 68(1):125-132). I work with the 50 micron thick vibratome sections of mouse brain embedded in agar which for me works better then gelatin. Normally, for flat embedding I osmicate and/or dehydrate the vibratome sections placed between two membrane filters disks with diameter at least two-three times exceeding the tissue size. The sections remained flat between two membrane disks due to the surface tension. The severe section distortion during dehydration is avoided or at least minimized.
The main disadvantage is the increased number of fluid changes and extended dehydration times compared to routine dehydration protocols, which makes sample preparation relatively time consuming. Briefly: place disk of filter paper into wetted compartment of Polystyrene multidish plate, transfer fixed vibratome sections onto filter disk and cover with the other one. Carefully add the solution not allowing the top filter to float. The challenge is to keep a certain minimal solution volume to maintain a surface tension between filters and yet sufficient to avoid sample drying. I not always use propylene oxide when working with TAAB embedding and the polystyrene plate works well with ethanol. For the last 100% change I add alcohol in excess to remove top filter disc without breaking the section. Aclar or Kapton films sandwich works very well for the polymerization Hope this helps, Albina
On Sat Jul 01 06:45:28 EDT 2006, dyel-at-mail.nih.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 } ---------------------------------------------------------------------------- } } 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 dyel-at-mail.nih.gov as well as the MIcroscopy } Listserver } --------------------------------------------------------------------------- } } Email: dyel-at-mail.nih.gov } Name: Chip Dye } } Organization: NIH } } Title-Subject: [Filtered] Flat embedding } } Question: Dear ListServers: } } Would anyone have any recommendations on the best way to flat } embed mouse embryos (12 days old) which have been sectioned on a } vibratome? I embedded the embryos in 10% gelatin prior to } sectioning and cut 50 micron sections and embedded in Epon. It } was during dehydration and infiltration, that my sections started } to curl. After polymerizing in a flat mold, my sections looked } like curled "potato chips". Would cutting the sections thicker } help with this issue? Any assistance as to the best method to } keep sections from doing this, would be greatly appreciated. } } Regards, } } Chip Dye } } Microscopist } Microscopy & Imaging Core, NICHD, NIH } Building 49, Room 5W-14 } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } Phone: 301-496-3627 } E-mail: dyel-at-mail.nih.gov } } } } --------------------------------------------------------------------------- } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Sat Jul 1 05:34:49 2006 } 12, 12 -- Received: from [192.169.1.141] (msdvpn24.msd.anl.gov } [130.202.238.88]) } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k61AYk3B004150 } 12, 12 -- for {microscopy-at-microscopy.com} ; Sat, 1 Jul 2006 } 05:34:48 -0500 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p06020401c0cc003743f2-at-[192.169.1.141]} } 12, 12 -- Date: Sat, 1 Jul 2006 05:34:46 -0500 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: dyel-at-mail.nih.gov (by way of } MicroscopyListserver) } 12, 12 -- Subject: viaWWW: Flat embedding } 12, 12 -- Content-Type: text/plain; charset="us-ascii" ; } format="flowed" } ==============================End of - } Headers============================== } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 8, 22 -- From amich-at-ufl.edu Wed Jul 5 15:49:07 2006 8, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65Kn6EB031660 8, 22 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 15:49:07 -0500 8, 22 -- Received: from osgjas03.cns.ufl.edu (osgjas03.cns.ufl.edu [128.227.74.133]) 8, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k65Kn0NW4886700; 8, 22 -- Wed, 5 Jul 2006 16:49:01 -0400 8, 22 -- Message-ID: {2028944878.112041152132540772.JavaMail.osg-at-osgjas03.cns.ufl.edu} 8, 22 -- Date: Wed, 5 Jul 2006 16:49:00 -0400 (EDT) 8, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 8, 22 -- To: dyel-at-mail.nih.gov 8, 22 -- Subject: Re: [Microscopy] viaWWW: Flat embedding 8, 22 -- Cc: microscopy-at-microscopy.com 8, 22 -- MIME-Version: 1.0 8, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 8, 22 -- Content-Transfer-Encoding: 7bit 8, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 8, 22 -- X-Originating-IP: 70.152.43.35 [70.152.43.35] 8, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 8, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Ritchie, Geoff and others: for some odd reason my email program is thinking there is a space when there isn't one, so the link is not finished. If you copy and paste the URL into your browser it seems to work. Check to see if there is a space in the copy before you press 'enter'. There should be NO spaces in the URL.
Ritchie Sims wrote: } Hi } } Still doesn't work, and looks the same, did you do the same mis-poke? } } please try again } } cheers } } rtch } } } } On 5 Jul 2006 at 14:51, r-holdford-at-ti.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 } } ---------------------------------------------------------------------- } } ------ } } } } All: the URL I previous sent has no spaces in it. Must have been a } } finger mis-poke. } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } able/ } } } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 23 -- From r-holdford-at-ti.com Wed Jul 5 15:52:30 2006 4, 23 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65KqUUj004901 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 15:52:30 -0500 4, 23 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 23 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k65Kq75D016380; 4, 23 -- Wed, 5 Jul 2006 15:52:12 -0500 (CDT) 4, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 23 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k65Kq6Pu021536; 4, 23 -- Wed, 5 Jul 2006 15:52:06 -0500 (CDT) 4, 23 -- Message-ID: {44AC2676.1000800-at-ti.com} 4, 23 -- Date: Wed, 05 Jul 2006 15:52:06 -0500 4, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 23 -- Organization: SC Packaging Development -- FA Development 4, 23 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 4, 23 -- MIME-Version: 1.0 4, 23 -- To: Ritchie Sims {r.sims-at-auckland.ac.nz} , mcauliff-at-umdnj.edu, 4, 23 -- MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 23 -- Subject: Re: [Microscopy] Re: need x-ray emission table 4, 23 -- References: {44ACC831.12352.4E962-at-localhost} 4, 23 -- In-Reply-To: {44ACC831.12352.4E962-at-localhost} 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Just as an FYI, the first and second tries worked fine for me... I didn't have any problem with the url sent.
dj
On Wed, 5 Jul 2006, r-holdford-at-ti.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 } ---------------------------------------------------------------------------- } } Ritchie, Geoff and others: for some odd reason my email program is } thinking there is a space when there isn't one, so the link is not } finished. If you copy and paste the URL into your browser it seems to } work. Check to see if there is a space in the copy before you press } 'enter'. There should be NO spaces in the URL. } } Ritchie Sims wrote: } } Hi } } } } Still doesn't work, and looks the same, did you do the same mis-poke? } } } } please try again } } } } cheers } } } } rtch } } } } } } } } On 5 Jul 2006 at 14:51, r-holdford-at-ti.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 } } } ---------------------------------------------------------------------- } } } ------ } } } } } } All: the URL I previous sent has no spaces in it. Must have been a } } } finger mis-poke. } } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } } able/ } } } } } } } } } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-995-2360 } 972-648-8743 (pager) } SC Packaging FA Development } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } ==============================Original Headers============================== } 4, 23 -- From r-holdford-at-ti.com Wed Jul 5 15:52:30 2006 } 4, 23 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) } 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65KqUUj004901 } 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 15:52:30 -0500 } 4, 23 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) } 4, 23 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k65Kq75D016380; } 4, 23 -- Wed, 5 Jul 2006 15:52:12 -0500 (CDT) } 4, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) } 4, 23 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k65Kq6Pu021536; } 4, 23 -- Wed, 5 Jul 2006 15:52:06 -0500 (CDT) } 4, 23 -- Message-ID: {44AC2676.1000800-at-ti.com} } 4, 23 -- Date: Wed, 05 Jul 2006 15:52:06 -0500 } 4, 23 -- From: Becky Holdford {r-holdford-at-ti.com} } 4, 23 -- Organization: SC Packaging Development -- FA Development } 4, 23 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) } 4, 23 -- MIME-Version: 1.0 } 4, 23 -- To: Ritchie Sims {r.sims-at-auckland.ac.nz} , mcauliff-at-umdnj.edu, } 4, 23 -- MSA ListServer {Microscopy-at-MSA.Microscopy.Com} } 4, 23 -- Subject: Re: [Microscopy] Re: need x-ray emission table } 4, 23 -- References: {44ACC831.12352.4E962-at-localhost} } 4, 23 -- In-Reply-To: {44ACC831.12352.4E962-at-localhost} } 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 23 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 5, 19 -- From dljones-at-bestweb.net Wed Jul 5 16:01:40 2006 5, 19 -- Received: from vms048pub.verizon.net (vms048pub.verizon.net [206.46.252.48]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65L1erQ019493 5, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 16:01:40 -0500 5, 19 -- Received: from localhost ([70.107.112.47]) 5, 19 -- by vms048.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 5, 19 -- 9 2005)) with ESMTPA id {0J1Y00CB77Q24RD9-at-vms048.mailsrvcs.net} for 5, 19 -- Microscopy-at-microscopy.com; Wed, 05 Jul 2006 16:01:19 -0500 (CDT) 5, 19 -- Date: Wed, 05 Jul 2006 17:12:32 -0400 (Eastern Standard Time) 5, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 5, 19 -- Subject: Re: [Microscopy] need x-ray emission table 5, 19 -- In-reply-to: {200607052056.k65KuXxo016061-at-ns.microscopy.com} 5, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 5, 19 -- To: r-holdford-at-ti.com 5, 19 -- Cc: Microscopy-at-microscopy.com 5, 19 -- Message-id: {Pine.WNT.4.64.0607051711330.4076-at-H-F1} 5, 19 -- MIME-version: 1.0 5, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 5, 19 -- References: {200607052056.k65KuXxo016061-at-ns.microscopy.com} ==============================End of - Headers==============================
The fatty and relatively soft brain tissue I embed in 3- 5% low melting point agarose (Sigma Chemical Co., cat# A-9414). I prefer not to trim the block too close to tissue: it seems that excess agarose helping prevent sections from curling.
} What percentage agar do you use when making the brain sections? } Is this regular bacteriological agar or low melting agarose? I } am about to embark on a similar project. I was thinking of } putting the sections into mesh biopsy bags to keep them flat. } } amich-at-ufl.edu wrote: } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } The issue of flat-embedding has been addressed by several } } authors, including Schwartz (1982), Schafer (1989), Nguyen and } } Pender (1995), Larue and Winer (1996). In my processing I } } largely follow the approach established by Larue and Winer } } (1996) with some modifications (Larue DT, Winer JA. } } Postembedding immunocytochemistry of large sections of brain } } tissue: an improved flat embedding technique. J Neurosci } } Methods, 1996 Sep; 68(1):125-132). I work with the 50 micron } } thick vibratome sections of mouse brain embedded in agar which } } for me works better then gelatin. Normally, for flat embedding I } } osmicate and/or dehydrate the vibratome sections placed between } } two membrane filters disks with diameter at least two-three } } times exceeding the tissue size. The sections remained flat } } between two membrane disks due to the surface tension. The } } severe section distortion during dehydration is avoided or at } } least minimized. } } } } The main disadvantage is the increased number of fluid changes } } and extended dehydration times compared to routine dehydration } } protocols, which makes sample preparation relatively time } } consuming. Briefly: place disk of filter paper into wetted } } compartment of Polystyrene multidish plate, transfer fixed } } vibratome sections onto filter disk and cover with the other } } one. Carefully add the solution not allowing the top filter to } } float. The challenge is to keep a certain minimal solution } } volume to maintain a surface tension between filters and yet } } sufficient to avoid sample drying. I not always use propylene } } oxide when working with TAAB embedding and the polystyrene plate } } works well with ethanol. For the last 100% change I add alcohol } } in excess to remove top filter disc without breaking the section. } } Aclar or Kapton films sandwich works very well for the } } polymerization } } Hope this helps, } } Albina } } } } On Sat Jul 01 06:45:28 EDT 2006, dyel-at-mail.nih.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 } } } ---------------------------------------------------------------------------- } } } } } } 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 dyel-at-mail.nih.gov as well as the } } } MIcroscopy Listserver } } } --------------------------------------------------------------------------- } } } } } } Email: dyel-at-mail.nih.gov } } } Name: Chip Dye } } } } } } Organization: NIH } } } } } } Title-Subject: [Filtered] Flat embedding } } } } } } Question: Dear ListServers: } } } } } } Would anyone have any recommendations on the best way to flat } } } embed mouse embryos (12 days old) which have been sectioned on } } } a vibratome? I embedded the embryos in 10% gelatin prior to } } } sectioning and cut 50 micron sections and embedded in Epon. It } } } was during dehydration and infiltration, that my sections } } } started to curl. After polymerizing in a flat mold, my } } } sections looked like curled "potato chips". Would cutting the } } } sections thicker help with this issue? Any assistance as to } } } the best method to keep sections from doing this, would be } } } greatly appreciated. } } } } } } Regards, } } } } } } Chip Dye } } } } } } Microscopist } } } Microscopy & Imaging Core, NICHD, NIH } } } Building 49, Room 5W-14 } } } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } } } Phone: 301-496-3627 } } } E-mail: dyel-at-mail.nih.gov } } } } } } } } } } } } --------------------------------------------------------------------------- } } } ==============================Original } } } Headers============================== } } } 12, 12 -- From zaluzec-at-microscopy.com Sat Jul 1 05:34:49 2006 } } } 12, 12 -- Received: from [192.169.1.141] (msdvpn24.msd.anl.gov } } } [130.202.238.88]) } } } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } } ESMTP id k61AYk3B004150 } } } 12, 12 -- for {microscopy-at-microscopy.com} ; Sat, 1 Jul 2006 } } } 05:34:48 -0500 } } } 12, 12 -- Mime-Version: 1.0 } } } 12, 12 -- X-Sender: (Unverified) } } } 12, 12 -- Message-Id: {p06020401c0cc003743f2-at-[192.169.1.141]} } } } 12, 12 -- Date: Sat, 1 Jul 2006 05:34:46 -0500 } } } 12, 12 -- To: microscopy-at-microscopy.com } } } 12, 12 -- From: dyel-at-mail.nih.gov (by way of } } } MicroscopyListserver) } } } 12, 12 -- Subject: viaWWW: Flat embedding } } } 12, 12 -- Content-Type: text/plain; charset="us-ascii" ; } } } format="flowed" } } } ==============================End of - } } } Headers============================== } } } } } } } } } } } } } } -- } } MIKHAYLOVA,ALBINA, PhD } } Post Doctoral Research Associate } } Materials Science and Engineering } } University of Florida } } } } ==============================Original } } Headers============================== } } 8, 22 -- From amich-at-ufl.edu Wed Jul 5 15:49:07 2006 } } 8, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu } } [128.227.74.149]) } } 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k65Kn6EB031660 } } 8, 22 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 } } 15:49:07 -0500 } } 8, 22 -- Received: from osgjas03.cns.ufl.edu } } (osgjas03.cns.ufl.edu [128.227.74.133]) } } 8, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id } } k65Kn0NW4886700; } } 8, 22 -- Wed, 5 Jul 2006 16:49:01 -0400 } } 8, 22 -- Message-ID: } } {2028944878.112041152132540772.JavaMail.osg-at-osgjas03.cns.ufl.edu} } } 8, 22 -- Date: Wed, 5 Jul 2006 16:49:00 -0400 (EDT) } } 8, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} } } 8, 22 -- To: dyel-at-mail.nih.gov } } 8, 22 -- Subject: Re: [Microscopy] viaWWW: Flat embedding } } 8, 22 -- Cc: microscopy-at-microscopy.com } } 8, 22 -- MIME-Version: 1.0 } } 8, 22 -- Content-Type: text/plain; format=flowed; } } charset=us-ascii } } 8, 22 -- Content-Transfer-Encoding: 7bit } } 8, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) } } 8, 22 -- X-Originating-IP: 70.152.43.35 [70.152.43.35] } } 8, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED } } 8, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, } } tests=ALL_TRUSTED } } 8, 22 -- X-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } 8, 22 -- X-UFL-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } ==============================End of - } } Headers============================== } } } } -- Greg Erdos } 5410 SE 185th Ave. } Micanopy, FL 32667 } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 7, 21 -- From amich-at-ufl.edu Wed Jul 5 21:10:02 2006 7, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k662A1pn001133 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 21:10:02 -0500 7, 21 -- Received: from osgjas03.cns.ufl.edu (osgjas03.cns.ufl.edu [128.227.74.133]) 7, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k6629vkq1925370; 7, 21 -- Wed, 5 Jul 2006 22:09:58 -0400 7, 21 -- Message-ID: {4879372.115621152151797443.JavaMail.osg-at-osgjas03.cns.ufl.edu} 7, 21 -- Date: Wed, 5 Jul 2006 22:09:57 -0400 (EDT) 7, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 7, 21 -- To: gwe-at-ufl.edu, microscopy-at-microscopy.com 7, 21 -- Subject: Re: [Microscopy] Re: viaWWW: Flat embedding 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 7, 21 -- X-Originating-IP: 68.214.118.120 [68.214.118.120] 7, 21 -- X-Spam-Status: hits=0, required=5, tests= 7, 21 -- X-UFL-Spam-Status: hits=0, required=5, tests= 7, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 7, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Has anybody ever Experimentally tried this inside Wehnelt gold coating and measured or noticed a difference in the false peak response as an increase or decrease in the peak-to-peak amplitude of the false peak as seen on an older SEM's CRT waveform? (All other factors being equal.)
Has anyone ever Experimentally tried it with a thick carbon coating and noticed the peak-peak amplitude response of the false peak?
Paul
At 09:07 AM 6/24/06 -0500, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 25 -- From beaurega-at-westol.com Thu Jul 6 01:58:48 2006 7, 25 -- Received: from smtp-gateway-1.winbeam.com (smtp-gateway-1.winbeam.com [64.84.97.66]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k666wmmI020291 7, 25 -- for {microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 01:58:48 -0500 7, 25 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 7, 25 -- by smtp-gateway-1.winbeam.com (8.13.1/8.12.8) with SMTP id k666whiT003030 7, 25 -- for {microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 02:58:45 -0400 7, 25 -- Received: (qmail 31079 invoked by uid 89); 6 Jul 2006 06:58:41 -0000 7, 25 -- Received: from pitts-69-72-13-120.dynamic-dialup.coretel.net (HELO millenium) (69.72.13.120) 7, 25 -- by mail.winbeam.com with SMTP; 6 Jul 2006 06:58:41 -0000 7, 25 -- Message-Id: {3.0.6.32.20060706030001.007d46b0-at-pop3.norton.antivirus} 7, 25 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 7, 25 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 7, 25 -- Date: Thu, 06 Jul 2006 03:00:01 -0400 7, 25 -- To: microscopy-at-microscopy.com 7, 25 -- From: Beaurega {beaurega-at-westol.com} 7, 25 -- Subject: Re: [Microscopy] What would happen if ...... sputter coating 7, 25 -- Wehnelt assembly 7, 25 -- Mime-Version: 1.0 7, 25 -- Content-Type: text/plain; charset="us-ascii" 7, 25 -- X-Winbeam-MailScanner-Information: smtp-gateway-1.winbeam.com - Please contact Technical Support for more information 7, 25 -- X-Winbeam-MailScanner: Found to be clean [smtp-gateway-1.winbeam.com] (courtesy of Winbeam) 7, 25 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, score=0, 7, 25 -- required 6, autolearn=disabled) 7, 25 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
I am seriously considering investing in a microwave for our cell monolayers processing for TEM. I would like to have some comments from people already using this technique (advantages/disadvantages, worth the investment...) and also an idea of the money I would have to spend for it.
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
We use microwaves as our standard processing method now, both for decreasing our processing times and for improved quality of ultrastructure. We find that microwave processing requires much less time than conventional processing and is less extractive of cell contents, as a rule.
Cell layers usually require less time than more bulky samples, but even so I would expect that you will save time using MW processing. A disadvantage is that MW requires the user to be there pretty much constantly, since the steps are so short that it's difficult to wander off and do other things while your specimens sit in solutions for 15 minutes to an hour. Another interesting effect is that you may notice less apparent contrast in your stained grids, probably due to less extraction of cellular constituents leading to more even staining across structures (i.e., staining still works fine, but there is more to stain, leading to less apparent difference between adjacent components.)
Expense can be an issue, since a programmable laboratory microwave with variable wattage (almost a necessity these days), a vacuum chamber (highly recommended), a circulating water bath to prevent hot spots (really highly recommended) can set you back over $10,000US (about 8000 euros or so?). One could do MW processing with an ordinary kitchen microwave, using water and ice loads and neon bulbs to track hot spots, but that is, like, so last century. Not to mention far less repeatable and very inconvenient.
My feeling is that once you go MW, you will not go back, at least for the majority of your samples.
Hope this helps. Feel free to contact me if you have questions.
All the best, Randy
Randy Tindall 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
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Thursday, July 06, 2006 4:51 AM To: Tindall, Randy D.
Dear listers,
I am seriously considering investing in a microwave for our cell monolayers processing for TEM. I would like to have some comments from people already using this technique (advantages/disadvantages, worth the investment...) and also an idea of the money I would have to spend for it.
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Does anyone know where I can obtain an older model Edwards Pirani gauge part number PRCT10-P?
Thank you in advance,
Mick Thomas
------------------------------------------------- Mick Thomas UHV-STEM / FESEM Laboratory E-1 and F-3 Clark Hall (Lab) 212 Clark Hall (mail) Cornell University Ithaca, NY 14853
This is a very useful little program ... But more useful if it also included absrption edges.
genuinely :o) michael shaffer
SEM/MLA Research Coordinator Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
} All: the URL I previous sent has no spaces in it. Must have } been a finger mis-poke. } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTable/ } } r-holdford-at-ti.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 } } } ---------------------------------------------------------------------- } } ------ } } } } Debby and anyone else who wants it: here's a link to } download the files: } } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } able/ } } } } Debby Sherman wrote: } } } } } Gee..would you mind sharing that program with others? I } would love } } } to have a copy. } } } } } } Debby } } } } } } Debby Sherman, Manager Phone: 765-494-6666 } } } Life Science Microscopy Facility FAX: 765-494-5896 } } } Purdue University E-mail: dsherman-at-purdue.edu } } } S-052 Whistler Building } } } 170 S. University Street } } } West Lafayette, IN 47907 } } } http://www.agriculture.purdue.edu/microscopy } } } } } } } } } On 6/28/06 6:37 PM, "r-holdford-at-ti.com" {r-holdford-at-ti.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 } } } } } -------------------------------------------------------------------- } } } } -------- } } } } } } } } Owen: one of my favorites is EM Periodic Table, a } program authored } } } } by Scott Walck many years ago. It has a periodic chart } layout and a } } } } table layout that can be translated to Excel. I got it } at a Lehigh } } } } short course and I find it very handy. I'd be happy to } share it with you. } } } } Scott calls this "beerware" meaning when we meet him at } conferences, } } } } we owe him a beer if we use it. } } } } } } } } Scott: you can claim your pitcher at M&M in Chicago. } This program } } } } would be cheap at twice the price. } } } } } } } } opmills-at-mtu.edu wrote: } } } } } } } } } } } } } } ------------------------------------------------------------------- } } } } } --------- The Microscopy ListServer -- CoSponsor: The } Microscopy } } } } } Society of America To Subscribe/Unsubscribe -- } } } } } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help
==============================Original Headers============================== 5, 20 -- From Michael-at-Shaffer.net Thu Jul 6 12:07:14 2006 5, 20 -- Received: from ws6-1.us4.outblaze.com (ws6-1.us4.outblaze.com [205.158.62.196]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k66H7D2Q010236 5, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 12:07:13 -0500 5, 20 -- Received: (qmail 27077 invoked from network); 6 Jul 2006 17:07:09 -0000 5, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 5, 20 -- by ws6-1.us4.outblaze.com with SMTP; 6 Jul 2006 17:07:08 -0000 5, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 5, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 20 -- Subject: RE: [Microscopy] Re: need x-ray emission table 5, 20 -- Date: Thu, 6 Jul 2006 14:37:07 -0230 5, 20 -- Message-ID: {000a01c6a11e$9d7e0390$8d829986-at-roamingwolf} 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="us-ascii" 5, 20 -- Content-Transfer-Encoding: 7bit 5, 20 -- X-Mailer: Microsoft Office Outlook 11 5, 20 -- In-Reply-To: {200607051950.k65JonqE021573-at-ns.microscopy.com} 5, 20 -- Thread-Index: AcagbFPUO3EWLVX2ScWRsy+z/ZarYgAsfvoQ 5, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
Michael, The program does have the absorption edges in it. That's what an EELS line corresponds to. Check out Si for example. You will find the K-a line for Si at 1.74 in the XEDS mode. Then go to EELS mode and click on Si again and you will see that the Si K edge is 1839 eV.
BTW, to address the original question, the data files that come with the program are CSV (comma separated values) files and will open directly into Excel or another spreadsheet program. Two are XEDS data, one in keV and one it eV. The other file is the EELS data only and is only in eV. Before you copy it to use it, please look at the credits to Nestor, Noran, and EmiSpec for the data in the help menu of the program.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: michael-at-Shaffer.net [mailto:michael-at-Shaffer.net] Sent: Thursday, July 06, 2006 10:12 AM To: Walck-at-SouthBayTech.com
This is a very useful little program ... But more useful if it also included absrption edges.
genuinely :o) michael shaffer
SEM/MLA Research Coordinator Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
} All: the URL I previous sent has no spaces in it. Must have } been a finger mis-poke. } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTab le/ } } r-holdford-at-ti.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 } } } ---------------------------------------------------------------------- } } ------ } } } } Debby and anyone else who wants it: here's a link to } download the files: } } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } able/ } } } } Debby Sherman wrote: } } } } } Gee..would you mind sharing that program with others? I } would love } } } to have a copy. } } } } } } Debby } } } } } } Debby Sherman, Manager Phone: 765-494-6666 } } } Life Science Microscopy Facility FAX: 765-494-5896 } } } Purdue University E-mail: dsherman-at-purdue.edu } } } S-052 Whistler Building } } } 170 S. University Street } } } West Lafayette, IN 47907 } } } http://www.agriculture.purdue.edu/microscopy } } } } } } } } } On 6/28/06 6:37 PM, "r-holdford-at-ti.com" {r-holdford-at-ti.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 } } } } } -------------------------------------------------------------------- } } } } -------- } } } } } } } } Owen: one of my favorites is EM Periodic Table, a } program authored } } } } by Scott Walck many years ago. It has a periodic chart } layout and a } } } } table layout that can be translated to Excel. I got it } at a Lehigh } } } } short course and I find it very handy. I'd be happy to } share it with you. } } } } Scott calls this "beerware" meaning when we meet him at } conferences, } } } } we owe him a beer if we use it. } } } } } } } } Scott: you can claim your pitcher at M&M in Chicago. } This program } } } } would be cheap at twice the price. } } } } } } } } opmills-at-mtu.edu wrote: } } } } } } } } } } } } } } ------------------------------------------------------------------- } } } } } --------- The Microscopy ListServer -- CoSponsor: The } Microscopy } } } } } Society of America To Subscribe/Unsubscribe -- } } } } } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help
==============================Original Headers============================== 5, 20 -- From Michael-at-Shaffer.net Thu Jul 6 12:07:14 2006 5, 20 -- Received: from ws6-1.us4.outblaze.com (ws6-1.us4.outblaze.com [205.158.62.196]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k66H7D2Q010236 5, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 12:07:13 -0500 5, 20 -- Received: (qmail 27077 invoked from network); 6 Jul 2006 17:07:09 -0000 5, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 5, 20 -- by ws6-1.us4.outblaze.com with SMTP; 6 Jul 2006 17:07:08 -0000 5, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 5, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 20 -- Subject: RE: [Microscopy] Re: need x-ray emission table 5, 20 -- Date: Thu, 6 Jul 2006 14:37:07 -0230 5, 20 -- Message-ID: {000a01c6a11e$9d7e0390$8d829986-at-roamingwolf} 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="us-ascii" 5, 20 -- Content-Transfer-Encoding: 7bit 5, 20 -- X-Mailer: Microsoft Office Outlook 11 5, 20 -- In-Reply-To: {200607051950.k65JonqE021573-at-ns.microscopy.com} 5, 20 -- Thread-Index: AcagbFPUO3EWLVX2ScWRsy+z/ZarYgAsfvoQ 5, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
==============================Original Headers============================== 16, 24 -- From walck-at-southbaytech.com Thu Jul 6 13:27:01 2006 16, 24 -- Received: from ylpvm01.prodigy.net (ylpvm01-ext.prodigy.net [207.115.57.32]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k66IR0fZ021802 16, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 13:27:00 -0500 16, 24 -- X-ORBL: [64.169.193.90] 16, 24 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 16, 24 -- by ylpvm01.prodigy.net (8.13.7 out spool5000 dk/8.13.7) with ESMTP id k66IQuY5000387; 16, 24 -- Thu, 6 Jul 2006 14:26:58 -0400 16, 24 -- From: "Scott Walck" {walck-at-southbaytech.com} 16, 24 -- To: {michael-at-Shaffer.net} 16, 24 -- Cc: {Microscopy-at-microscopy.com} 16, 24 -- Subject: RE: [Microscopy] need x-ray emission table 16, 24 -- Date: Thu, 6 Jul 2006 11:27:46 -0700 16, 24 -- Message-ID: {003201c6a129$e232a9e0$7801a8c0-at-dynamicbl8uno3} 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-Type: text/plain; 16, 24 -- charset="us-ascii" 16, 24 -- Content-Transfer-Encoding: 7bit 16, 24 -- X-Priority: 3 (Normal) 16, 24 -- X-MSMail-Priority: Normal 16, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 16, 24 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 16, 24 -- Importance: Normal 16, 24 -- In-Reply-To: {200607061712.k66HCE9x016726-at-ns.microscopy.com} ==============================End of - Headers==============================
I am looking for a side mounted camera assembly for a FEI Tecnai 12. Philips CM or 400 series TEM. While a complete assembly, including camera, would be ideal, I have some digital cameras I can mount if I can get the ports and the yag/prism sidearm assembly.
Please contact me offline in this regard if you have a unit you are willing to part with
Steve
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu
==============================Original Headers============================== 6, 18 -- From sbarlow-at-sunstroke.sdsu.edu Thu Jul 6 14:32:17 2006 6, 18 -- Received: from sciences.sdsu.edu (sciences.sdsu.edu [130.191.140.33]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k66JWH49000947 6, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 14:32:17 -0500 6, 18 -- Received: from [146.244.225.122] ([146.244.225.122]) 6, 18 -- by sciences.sdsu.edu (8.12.11.20060614/8.12.10/SCEC-032005-9-50) with ESMTP id k66JWFDp026671 6, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 12:32:16 -0700 (PDT) 6, 18 -- Mime-Version: 1.0 6, 18 -- Message-Id: {p06110405c0d3154fc4d8-at-[146.244.225.122]} 6, 18 -- Date: Thu, 6 Jul 2006 12:32:08 -0700 6, 18 -- To: Microscopy-at-microscopy.com 6, 18 -- From: Steve Barlow {sbarlow-at-sunstroke.sdsu.edu} 6, 18 -- Subject: (TEM) used side mounted camera assembly 6, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 18 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-1.6 (sciences.sdsu.edu [130.191.140.35]); Thu, 06 Jul 2006 12:32:16 -0700 (PDT) 6, 18 -- X-COS-MailScanner: Found to be clean 6, 18 -- X-MailScanner-SpamCheck: not spam, SpamAssassin (score=0, required 8) 6, 18 -- X-MailScanner-From: sbarlow-at-sunstroke.sdsu.edu ==============================End of - Headers==============================
Listers, I am trying to fix an old LBK IV ultramicrotome Model 2128. The metal band that connects the drive motor to the specimen arm has broken. I can probably carry out the repairs myself but need to locate a replacement part. I think that there is no official parts source these days but wondered if anyone had already cannibalized such an instrument or had an unused one suitable for spares. Any help much appreciated.
Best regards
Chris
Christopher J Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.A04 UT Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-9039 Phone +1 214 648 2827 Fax +1 214 648 6408
==============================Original Headers============================== 6, 21 -- From christopher.gilpin-at-utsouthwestern.edu Thu Jul 6 17:43:51 2006 6, 21 -- Received: from swlx166.swmed.edu (swlx166.swmed.edu [199.165.152.166]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k66MhpKo014100 6, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 17:43:51 -0500 6, 21 -- Received: from [129.112.148.58] (helo=cgdesktop) 6, 21 -- by swlx166.swmed.edu with esmtp (Exim 4.44) 6, 21 -- id 1FycZZ-0002YK-Kh 6, 21 -- for Microscopy-at-microscopy.com; Thu, 06 Jul 2006 17:43:51 -0500 6, 21 -- From: "Christopher Gilpin" {christopher.gilpin-at-utsouthwestern.edu} 6, 21 -- To: {Microscopy-at-microscopy.com} 6, 21 -- Date: Thu, 6 Jul 2006 17:43:58 -0500 6, 21 -- Message-ID: {001301c6a14d$ab4d2030$3a947081-at-cgdesktop} 6, 21 -- MIME-Version: 1.0 6, 21 -- Content-Type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-Transfer-Encoding: 7bit 6, 21 -- X-Mailer: Microsoft Office Outlook 11 6, 21 -- Thread-Index: AcahTasR/3FlEY2zSBiQQoQxao6KCA== 6, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 6, 21 -- X-Scan-Signature: caec9e7ca10ae3625b33f2c31db97675 6, 21 -- Subject: Parts and repair for LKB IV ultramicrotome ==============================End of - Headers==============================
It gets pretty hot near the filament. I suspect that the sputtered on coating would flake off due to the differential thermal expansion of the two metals since conventionally sputtered metals are sitting lightly on the surface. On the other hand, electroplated metals would be more stable. For example, we have an ancient Wehnelt from an RCA that is gold plated. It was stable for 30 or more years. BUT, then, apparently, someone tried to clean it too vigorously and removed some of the gold. Now the cleaned area is tarnished.
Perhaps Steve Chapman and Chuck Garber might want to chime in here as they are experienced in scope maintenance and metalurgy, respectively.
JB -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 4, 16 -- From bozzola-at-siu.edu Thu Jul 6 20:57:31 2006 4, 16 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 4, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k671vVKk026946 4, 16 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 6 Jul 2006 20:57:31 -0500 4, 16 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 4, 16 -- by abbmta2.siu.edu (Switch-3.1.9/Switch-3.1.7) with ESMTP id k671vT7A004272 4, 16 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 6 Jul 2006 20:57:30 -0500 (CDT) 4, 16 -- Mime-Version: 1.0 4, 16 -- X-Sender: bozzola-at-saluki-mail.siu.edu 4, 16 -- Message-Id: {p0611040bc0d36d89bfad-at-[131.230.177.142]} 4, 16 -- Date: Thu, 6 Jul 2006 20:57:31 -0500 4, 16 -- To: Microscopy-at-msa.microscopy.com 4, 16 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 4, 16 -- Subject: Re: What would happen if ...... sputter coating 4, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 16 -- X-MASF: 0.00% ==============================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 shapiro-at-unbc.ca as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: shapiro-at-unbc.ca Name: Aaron Shapiro
Organization: University of Northern British Columbia
Title-Subject: [Filtered] Human CNS Atlas
Question: I was wondering if anyone was aware of a good histology atlas which shows TEM images of human or rat central nervous system - particularly the granule cell layer of the cerebellum.
you can also try MA-Table for X-ray emission lines, wisible with EDS. There are more lines in L- and M-series than in other data basis. This is the manual and there you'll find also the download: http://microanalyst.mikroanalytik.de/manual.html
Regards
Frank
opmills-at-mtu.edu schrieb:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 19 -- From eggert-at-mikroanalytik.de Fri Jul 7 00:57:32 2006 7, 19 -- Received: from Mail1.KONTENT.De (mail1.kontent.de [81.88.34.36]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k675vW2f026780 7, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 00:57:32 -0500 7, 19 -- Received: from mikroanalytik.de (p54BDD829.dip.t-dialin.net [84.189.216.41]) 7, 19 -- by Mail1.KONTENT.De (Postfix) with ESMTP id 290C3108FAFB; 7, 19 -- Fri, 7 Jul 2006 07:57:33 +0200 (CEST) 7, 19 -- Message-ID: {44ADF8B4.7090102-at-mikroanalytik.de} 7, 19 -- Date: Fri, 07 Jul 2006 08:01:24 +0200 7, 19 -- From: Frank Eggert {eggert-at-mikroanalytik.de} 7, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; de-DE; rv:1.4) Gecko/20030619 Netscape/7.1 (ax) 7, 19 -- X-Accept-Language: de, de-de, en 7, 19 -- MIME-Version: 1.0 7, 19 -- To: opmills-at-mtu.edu, Microscopy-at-microscopy.com 7, 19 -- Subject: Re: [Microscopy] need x-ray emission table 7, 19 -- References: {200606282002.k5SK2YYY024728-at-ns.microscopy.com} 7, 19 -- In-Reply-To: {200606282002.k5SK2YYY024728-at-ns.microscopy.com} 7, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
We are happy to announce the first release of the 2dx software.
2dx provides a graphical user interface for the computer processing of 2D crystal images. So far, the program 2dx_image is available for the processing of individual images of tilted or non-tilted 2D crystals. Merging will be implemented in the near future.
2dx assists running c-shell scripts that call other stand-alone programs. In the current release we provide a set of MRC programs, which were slightly adapted to interact with the 2dx software, and we added a few functions trying to implement automatic determination of all required parameters. It is easy to adapt or replace the provided scripts with your own scripts and/or to make use of other back-end programs (Spider, bsoft, etc.) or your own MRC software.
2dx including the source codes is freely available, and we provide compiled versions for Linux x86_64bit and Mac-PPC (G4 (slow), G5) and the new Mac-Intel computers, see here: http://2dx.org The installer hopefully take care of everything (all goes into /usr/ local/2dx), and installs also the required modified MRC programs. The only thing that is required in addition is CCP4, as described on the server. But CCP4 is only used in the last script for the generation of a map. The rest should also work without CCP4. On the server are also some test data, in MRC-image2000 file format for big and small endian machines, together with the corresponding config files (2dx_image.cfg).
A manuscript that describes this software is under review at JSB, but we don't have written the manual for 2dx yet. However, hopefully, most features in 2dx are self-explanatory. Just try the right mouse button over all the different items, or double-clicking.
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 AIM:HStahlberg-at-mac.com
Workshop on Electron Crystallography, August 7-11, 2006, at UC Davis. http://2dx.org/workshop
2dx Software Version 1.0.0 now Available http://2dx.org/download/2dx-software/ _____________________________________________________________
==============================Original Headers============================== 18, 16 -- From HStahlberg-at-ucdavis.edu Fri Jul 7 15:40:07 2006 18, 16 -- Received: from pop23.ucdavis.edu (pop23.ucdavis.edu [169.237.105.13]) 18, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k67Ke7ol026329 18, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 15:40:07 -0500 18, 16 -- Received: from [169.237.214.65] ([169.237.214.65]) 18, 16 -- by pop23.ucdavis.edu (8.13.6/8.13.1/it-std-5.2.0) with ESMTP id k67Ke7a7014362 18, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 13:40:07 -0700 (PDT) 18, 16 -- Mime-Version: 1.0 (Apple Message framework v752.2) 18, 16 -- Content-Transfer-Encoding: 7bit 18, 16 -- Message-Id: {EACF3DDD-9442-4698-98F9-9B64934B101C-at-ucdavis.edu} 18, 16 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 18, 16 -- To: Microscopy-at-microscopy.com 18, 16 -- From: Henning Stahlberg {HStahlberg-at-ucdavis.edu} 18, 16 -- Subject: 2dx - image processing for 2D membrane protein crystals 18, 16 -- Date: Fri, 7 Jul 2006 13:40:01 -0700 18, 16 -- X-Mailer: Apple Mail (2.752.2) ==============================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 gradecak-at-fas.harvard.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
I have a question regarding image-diffraction pattern rotation in TEM. While in some TEMs this rotation is compensated, it is still possible to have a 180deg rotation between a TEM image and the corresponding diffraction pattern. What are the procedures to measure this rotation with or without special calibration samples?
I have a user who wants to know how thick a metal coating has to be to clog up the pores in a nucleopore filter.
I know, why not just try different amounts until it works, but this is an engineer type who wants to know the real numbers.
For this project he is trying to restrict the number of open pores in the filter to just a few in the center of his 1 cm dia. filter. The plan to do this for now is to put a 1 mm x 1 mm mask on the filter and evaporate or sputter some metal over the rest.
We were thinking gold or gold/paladium, but if you have a better idea, we're game for that.
The pores he wants to clog are 10 nm, 30 nm, 50 nm or 80 nm in a standard polycarbonate nucleopore filter. Objective is to stop fluid from going through the clogged pores, while still keeping the central 1 mm square pores open. The clogging stuff does not have to be electrically conductive, metal coating was just our first guess at a way to do it. Other strategies OK if they will work.
Thanks
Jon
--
Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 12, 21 -- From jmkrupp-at-ucsc.edu Fri Jul 7 17:38:22 2006 12, 21 -- Received: from smtp-prod-mx1.ucsc.edu (smtp-prod-mx1.ucsc.edu [128.114.125.43]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k67McLFb015788 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 17:38:21 -0500 12, 21 -- Received: from ucsc.edu (cruzmail-fe1.ucsc.edu [128.114.125.5]) 12, 21 -- by smtp-prod-mx1.ucsc.edu (8.13.7/8.13.7) with ESMTP id k67McH9r018876 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 15:38:17 -0700 (PDT) 12, 21 -- Received: from [128.114.25.141] (account jmkrupp-at-ucsc.edu [128.114.25.141] verified) 12, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 12, 21 -- with ESMTPA id 62079501 for microscopy-at-microscopy.com; Fri, 07 Jul 2006 15:38:17 -0700 12, 21 -- Mime-Version: 1.0 12, 21 -- Message-Id: {p06230906c0d4903dcf6c-at-[128.114.25.141]} 12, 21 -- Date: Fri, 7 Jul 2006 15:38:16 -0700 12, 21 -- To: microscopy-at-microscopy.com 12, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 12, 21 -- Subject: Thickness of metal coating needed 12, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 12, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 12, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 12, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 12, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
Go to convergent beam mode and go to cross over with the condenser lens. Then decrease the lens strength on the condenser (CCW) until you see the shadow image of the sample in the bright field disk. It should agree with the image orientation in the mag mode. (I think all microscopes have the knobs go clockwise to increase lens strength and CCW to decrease lens strength on the lenses.)
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: gradecak-at-fas.harvard.edu [mailto:gradecak-at-fas.harvard.edu] Sent: Friday, July 07, 2006 2:54 PM To: Walck-at-SouthBayTech.com
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 gradecak-at-fas.harvard.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
I have a question regarding image-diffraction pattern rotation in TEM. While in some TEMs this rotation is compensated, it is still possible to have a 180deg rotation between a TEM image and the corresponding diffraction pattern. What are the procedures to measure this rotation with or without special calibration samples?
We recently are having serious embedding problems using EPON generic (LX-112) formulation that we have used for years. Most of the problems seem to be in high pressure frozen samples including cultured mammalian cells and single cell cyanobacteria. Both samples embedded just fine a few months ago as did plant samples in the same resin formulation. Current cells are cryo-protected with dextran (~10% final concentration) and freezing looks great.
Cells themselves are well infiltrated but problems result in sections not spreading well and resin pulling away from cell walls (cyanobacteria). The samples go through a very long (5day) gradual infiltration, gentle agitation, and attention is paid to keeping everything dry, etc...all the things we have been doing for years with never a problem.
Any suggestions would be appreciated. I am about to revert back to Spurr resin even though we may have problems with the new VCD replacement. We went to the EPON formulation to get better contrast. Suggestions as to other resins or resin mixtures would also be appreciated.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 7, 21 -- From dsherman-at-purdue.edu Fri Jul 7 21:50:59 2006 7, 21 -- Received: from 1061exfe03.adpc.purdue.edu (1061exfe03.adpc.purdue.edu [128.210.63.225]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k682ox3o006266 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 21:50:59 -0500 7, 21 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe03.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 21 -- Fri, 7 Jul 2006 22:50:59 -0400 7, 21 -- Received: from 74.132.211.81 ([74.132.211.81]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 7, 21 -- Sat, 8 Jul 2006 02:50:56 +0000 7, 21 -- User-Agent: Microsoft-Entourage/11.2.4.060510 7, 21 -- Date: Fri, 07 Jul 2006 22:50:55 -0400 7, 21 -- Subject: Embedding problems 7, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 7, 21 -- Message-ID: {C0D495CF.3EC2%dsherman-at-purdue.edu} 7, 21 -- Thread-Topic: Embedding problems 7, 21 -- Thread-Index: AcaiOVUHk4GIvg4sEdu0zgAKlcoUxg== 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-OriginalArrivalTime: 08 Jul 2006 02:50:59.0628 (UTC) FILETIME=[57C93EC0:01C6A239] ==============================End of - Headers==============================
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Email: mdyousuf-at-qu.edu.qa Name: Mohammed Yousuf
Organization: Qatar University
Title-Subject: [Filtered] Philips CM12 - digital cameras
Question: Dear All, Please advice regarding purchase of a new digital camera for our CM12 scope. As of now the scope is in average health given the time that it has served since its instillation in 1992. My question is - is it adviseable to go in for a digital camera to this instrument? If yes, what are the options available out there. I need information from places who have had a recent instillation. Also need to know the price/ performance index for various options (individual brands/models and their resolution). Kindly respond to me direct.
Thanks in advance.
Mohammed Yousuf EM unit, Central Research Laboratories Qatar University P.O.Box 2713 Doha, State of Qatar mdyousuf-at-qu.edu.qa mdyousuf99-at-yahoo.com
This Question was submitted to Ask-A-Microscopist by (lasvegassierra-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, July 9, 2006 at 07:52:01 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lasvegassierra-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I found this site kinda by chance. I am excited at the possiblility of sharing some findings of mine. I also hope that maybe one of the scientist can help me to identify this thing. I dont know if anyone is willing to try or how I would send you a copy. Thru regular email is one thing but this program is different. Is it possible for you to view my microscopic views with me? Thank you for your time. Amie
I am a bit puzzled by your questions because you used methanol on other materials and it appeared to work for you. Anyway, the questions you asked are probably of general use. You asked: 1) does methanol evaporation cause particle agglomeration on the grid? I think you mean flocculation and not agglomeration. Yes, pure methanol does cause this problem and so do other solvents. It depends on the nature of your particles as far as the materials they are made up of and whether they are hydrophobic or hydrophilic as well. These are not the only variables by a long shot.
} 2) does methanol use cause morphological changes of the particles? Not to the primary particles or primary structural units, unless they dissolve in MeOH.
} 3) are particles size-fractionated on the TEM grid during the } evaporation process? Absolutely. Particles do not 'drop out' of solution by gravity onto a grid. The particles move around in a dynamic way. So they can be segregated. Some of this can be stopped to get good representative dispersions. Furthermore, dropping the particle suspension onto a grid on a piece of filter paper should be avoided unless you want to skew the size results to get a biased result. This is done in some patents and is a 'hidden' bias in one ASTM TEM method on carbon blacks. Flocculation on a grid can also combine aggregate structural units to make you think the 'particle sizes' are larger.
} 4) does methanol dissolve some phases? I guess the technical answer is yes, if the phase is even slightly soluble in methanol. I would not think this applies to your 'flyash' samples that should have things like glassy particles and unburned coal or carbon in them.
} My question to the Listserver is - should I avoid methanol? Not if you need a slightly polar solvent to disperse the particles of interest. } which solvents should I use and why? Try different solvent systems. Use the one you think works the best for your sample situation.
General comments: Don't get tunnel vision when dispersing powders, aggregates, or primary particles. Try different solvents and look at them in a TEM. Look at how they behave. Examine the whole grid to see the different patterns you see as a dried dispersion.
There's no magic bullet single dispersing system here. That's certainly true of colored ink jet pigment dispersions. The number of factors that come into play in dispersing powders is not a single dimensional variable called a solvent. Here's a pointed example of tunnel vision. I was told that a sample of some plasma generated nanoparticles was hydrophobic. The newly hired chemist's sonicated settling tests showed it would not disperse in water based systems. What would you use? I was told which organic solvent to use and I used that one. Then I dispersed the hydrophobic sample in a water based system. Both preps gave equivalent flocculation free dispersions on TEM grids. This was done by manipulating the other factors important in dispersion besides the type of solvent system.
In a previous posting from Leslie at IBM the following summary item was detailed. } There were two (who) mentioned dry loading the grid, which is dumping some of } the powder on the grid and tapping off the excess. This is the method I } tried first as it was the easiest, but I have not looked at the grid yet } to see if there is enough material to do EELS on. This (DeGussa-Germany?) method relies on the 'adhesion' of the finest aggregates to a grid film. They are attached simply by static charge. My experience with silica and plasma nanoparticles was that the populations were usually too light for me to use. The aggregate size distribution was skewed towards smaller sizes. Another problem is that the powder aggregates get on both sides of the grid without special handling precautions. This causes some aggregates on the wrong side of the grid, to be over focused. Yet, dry preps are faster to perform than solvent preps or mulling. I used dry preps when speed was critical, and the structure or size was not of interest.
Recently, the change in the size of ppt'd silica particles was blamed on a certain dispersing solvent. A dry prep would have shown that the size increased without that solvent ever being used. A dry prep is a good referee technique. For me, high speed high-res imaging of silica dry preps showed the presence of single digit nanometer scale salt on the surfaces of primary particles that accelerates this above mentioned "poached egg" look but it would happen even without the trace salts, given enough beam time. So dry preps have a place in a TEM lab and are useful.
Disclaimers: I performed dispersions on all types of powders and examined thin sections of the interiors of agglomerates, large aggregates, coatings of all types, and silica tire cleats for over 20 years. Your samples might perform differently than stated here. There are at least 10 factors or variables that apply to dispersions of powders and aggregates. That's too many to discuss here. Obtaining a good dispersion is a learned skill and takes practice.
Paul Beauregard Senior Research Associate Greensburg, PA 724-834-2247
At 09:10 PM 6/25/06 -0500, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 13, 24 -- From beaurega-at-westol.com Mon Jul 10 08:54:47 2006 13, 24 -- Received: from smtp-gateway-5.winbeam.com (smtp-gateway-5.winbeam.com [64.84.97.70]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6ADskA1016864 13, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Jul 2006 08:54:47 -0500 13, 24 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 13, 24 -- by smtp-gateway-5.winbeam.com (8.13.2/8.12.8) with SMTP id k6ADsTX9028600 13, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Jul 2006 09:54:29 -0400 13, 24 -- Received: (qmail 26830 invoked by uid 89); 10 Jul 2006 13:54:27 -0000 13, 24 -- Received: from pitts-69-72-110-171.dynamic-dialup.coretel.net (HELO millenium) (69.72.110.171) 13, 24 -- by mail.winbeam.com with SMTP; 10 Jul 2006 13:54:27 -0000 13, 24 -- Message-Id: {3.0.6.32.20060710095550.007f8560-at-pop3.norton.antivirus} 13, 24 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus (Unverified) 13, 24 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 13, 24 -- Date: Mon, 10 Jul 2006 09:55:50 -0400 13, 24 -- To: mgb-at-ansto.gov.au, Microscopy-at-microscopy.com, lkrupp-at-us.ibm.com 13, 24 -- From: Beaurega {beaurega-at-westol.com} 13, 24 -- Subject: Re: [Microscopy] TEM powder sample preparation 13, 24 -- Mime-Version: 1.0 13, 24 -- Content-Type: text/plain; charset="us-ascii" 13, 24 -- X-Winbeam-MailScanner-Information: - Please contact Technical Support for more information 13, 24 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) 13, 24 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, score=0, 13, 24 -- required 6, autolearn=disabled) 13, 24 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
One of our users is asking for references (books, articles etc.) concerning the accuracy of SEM measurements for particles. I explained to him the principles (difficulties with edge effect etc.), but he needs printed references for his article.
If anyone has an article name/book chapter, I would greatly appreciate it.
Thanks, Daniel Salamon Technical Officer, Electron Microscopy
National Institute for Nanotechnology, NRC 11421 Saskatchewan Dr. Edmonton, AB. T6G 2M9
Phone: (780) 641-1663 Fax: (780) 641-1601
==============================Original Headers============================== 7, 23 -- From Daniel.Salamon-at-nrc-cnrc.gc.ca Mon Jul 10 10:18:12 2006 7, 23 -- Received: from nrccenexf2.nrc.ca (nrccenexf2.nrc.ca [132.246.15.83]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6AFIBRM028823 7, 23 -- for {microscopy-at-microscopy.com} ; Mon, 10 Jul 2006 10:18:12 -0500 7, 23 -- Received: from nrccenexb2.nrc.ca ([132.246.15.98]) by nrccenexf2.nrc.ca with Microsoft SMTPSVC(6.0.3790.1830); 7, 23 -- Mon, 10 Jul 2006 11:18:06 -0400 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: SEM magnification accuracy references 7, 23 -- Date: Mon, 10 Jul 2006 11:18:05 -0400 7, 23 -- Message-ID: {923687253229634E96E808B8D3124C839789C3-at-nrccenexb2.nrc.ca} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: SEM magnification accuracy references 7, 23 -- Thread-Index: AcakNAquO+NWUArVRvi14ZEbnys6cQ== 7, 23 -- From: "Salamon, Daniel" {Daniel.Salamon-at-nrc-cnrc.gc.ca} 7, 23 -- To: {microscopy-at-microscopy.com} 7, 23 -- X-OriginalArrivalTime: 10 Jul 2006 15:18:06.0202 (UTC) FILETIME=[0B551DA0:01C6A434] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6AFIBRM028823 ==============================End of - Headers==============================
X-from cross over, under- or over-focusing the condenser lens will make both make it parallel. In the analytical machines that have more than two condenser lenses (as in the last 20+) years, you obtain the parallel condition faster (by less turns of the knob) by over-focusing the C2 lens than going under-focus. In machines that only have the two condenser system, you under focus the condenser to go to parallel condition faster. In either case, you never get to a parallel condition, you get to a very small convergence angle.
With respect to my previous answer, I believe that you can check the technique that I gave with a GaAs [011] CBED image since it is a non-centrosymmetric crystal.
As far as I know, TEM's still have the condition that CW and CCW still relate to the strength in the lens. You can watch the current or voltage monitor of the lenses when you turn the knob. Some manufacturer's of SEM's have changed that, especially with the objective lens. Now they relate the CW turn of the focus know with increasing the working distance, which of course, is decreasing the strength of the objective lens.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: Dusha [mailto:a.chuvilin-at-microscopist.ru] Sent: Sunday, July 09, 2006 8:36 AM To: walck-at-southbaytech.com
Don't to forget about dimensional calibration of the SEM. If this is not correct, all bets are off.
gary g.
At 08:20 AM 7/10/2006, you wrote:
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My first thought would be to use this filter exactly as he is intended to: to filter! Masking the central part, you can filter a suspension of particles which will eventually block the filter pores. I am confident that precipitates likes calcium carbonate or calcium oxalate would block the filter. Of course it all depends on the solution you have to filter (and its pH). Any particulate suspension will block the filter. Chemically neutral particles like fine mineral powder (silicate?) should work well too. It very much depends on what is available to you.
Regards,
Stephane
--- jmkrupp-at-ucsc.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi: } } I have a user who wants to know how thick a metal } coating has to be } to clog up the pores in a nucleopore filter. } } I know, why not just try different amounts until it } works, but this } is an engineer type who wants to know the real } numbers. } } For this project he is trying to restrict the number } of open pores in } the filter to just a few in the center of his 1 cm } dia. filter. The } plan to do this for now is to put a 1 mm x 1 mm mask } on the filter } and evaporate or sputter some metal over the rest. } } We were thinking gold or gold/paladium, but if you } have a better } idea, we're game for that. } } The pores he wants to clog are 10 nm, 30 nm, 50 nm } or 80 nm in a } standard polycarbonate nucleopore filter. Objective } is to stop fluid } from going through the clogged pores, while still } keeping the central } 1 mm square pores open. The clogging stuff does not } have to be } electrically conductive, metal coating was just our } first guess at a } way to do it. Other strategies OK if they will work. } } Thanks } } Jon } } } -- } } } Jonathan Krupp } Microscopy & Imaging Lab } C230 Earth & Marine Science } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-ucsc.edu } } ==============================Original } Headers============================== } 12, 21 -- From jmkrupp-at-ucsc.edu Fri Jul 7 17:38:22 } 2006 } 12, 21 -- Received: from smtp-prod-mx1.ucsc.edu } (smtp-prod-mx1.ucsc.edu [128.114.125.43]) } 12, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k67McLFb015788 } 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Jul 2006 17:38:21 -0500 } 12, 21 -- Received: from ucsc.edu } (cruzmail-fe1.ucsc.edu [128.114.125.5]) } 12, 21 -- by smtp-prod-mx1.ucsc.edu (8.13.7/8.13.7) } with ESMTP id k67McH9r018876 } 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Jul 2006 15:38:17 -0700 (PDT) } 12, 21 -- Received: from [128.114.25.141] (account } jmkrupp-at-ucsc.edu [128.114.25.141] verified) } 12, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP } 4.3.7) } 12, 21 -- with ESMTPA id 62079501 for } microscopy-at-microscopy.com; Fri, 07 Jul 2006 15:38:17 } -0700 } 12, 21 -- Mime-Version: 1.0 } 12, 21 -- Message-Id: } {p06230906c0d4903dcf6c-at-[128.114.25.141]} } 12, 21 -- Date: Fri, 7 Jul 2006 15:38:16 -0700 } 12, 21 -- To: microscopy-at-microscopy.com } 12, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} } 12, 21 -- Subject: Thickness of metal coating needed } 12, 21 -- Content-Type: text/plain; } charset="us-ascii" ; format="flowed" } 12, 21 -- X-UCSC-CATS-Information: This message was } scanned by the ITS MailScanner } 12, 21 -- X-UCSC-CATS-MailScanner: Found to be clean } 12, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: } 12, 21 -- X-UCSC-CATS-MailScanner-From: } jmkrupp-at-ucsc.edu } ==============================End of - } Headers============================== }
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We have been doing laser microdissection with MMI's (Molecular Machines & Industries) setup. We have been asked to do a sample for RNase analysis. MMI supplies RNase free membrane slides for the sections, but the collection tubes are not RNase free. Does anyone have a source for RNase free collection tubes for microdissection?
Mannie Steglich U T M D Anderson Cancer Center
==============================Original Headers============================== 2, 15 -- From msteglic-at-mdanderson.org Tue Jul 11 09:46:03 2006 2, 15 -- Received: from UTM-MAIL04A.mdacc.tmc.edu (utmlnmail04nt.mdacc.tmc.edu [143.111.84.152]) 2, 15 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6BEk22T028914 2, 15 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 11 Jul 2006 09:46:03 -0500 2, 15 -- To: Microscopy-at-MSA.Microscopy.Com 2, 15 -- Subject: RNase free Microdissection 2, 15 -- MIME-Version: 1.0 2, 15 -- X-Mailer: Lotus Notes Release 5.0.11 July 24, 2002 2, 15 -- Message-ID: {OF6E4AD7C7.00F49114-ON862571A8.0050B5F4-862571A8.00513D27-at-mdacc.tmc.edu} 2, 15 -- From: msteglic-at-mdanderson.org 2, 15 -- Date: Tue, 11 Jul 2006 09:46:01 -0500 2, 15 -- X-MIMETrack: Serialize by Router on UTM-MAIL04A/HOU/UTMDACC(Release 5.0.11 |July 24, 2002) at 2, 15 -- 07/11/2006 09:46:03 AM, 2, 15 -- Serialize complete at 07/11/2006 09:46:03 AM 2, 15 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Geoff- You are correct. The Sapphatomes were sapphire knives. They were supposed to be low-cost alternatives to diamonds. They cut well, but were almost impossible to keep clean, and did not wear nearly as well as hoped. Ages ago, we had one in the lab I was in at the time. It became the training knife for a new technician. Then it went into a drawer, never again to see the light of day. 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, 24 -- From lcgould-at-med.cornell.edu Tue Jul 11 14:28:57 2006 1, 24 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 1, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6BJSuU5021477 1, 24 -- for {microscopy-at-microscopy.com} ; Tue, 11 Jul 2006 14:28:56 -0500 1, 24 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 24 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k6BJSrNO026917 1, 24 -- for {microscopy-at-microscopy.com} ; Tue, 11 Jul 2006 15:28:54 -0400 (EDT) 1, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 24 -- by mpx1.med.cornell.edu 1, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 24 -- with ESMTPA id {0J29002Z07G5PL50-at-mpx1.med.cornell.edu} for 1, 24 -- microscopy-at-microscopy.com; Tue, 11 Jul 2006 15:28:53 -0400 (EDT) 1, 24 -- Date: Tue, 11 Jul 2006 15:20:58 -0400 1, 24 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 24 -- Subject: Re: [Microscopy] microtome knife question 1, 24 -- In-reply-to: {200607111913.k6BJDd9P012443-at-ns.microscopy.com} 1, 24 -- Sender: lcgould-at-med.cornell.edu 1, 24 -- To: Geoffrey_Williams-at-brown.edu, 1, 24 -- Microscopy Listserver {microscopy-at-microscopy.com} 1, 24 -- Message-id: {p06230914c0d9a9a1dfb4-at-[140.251.48.23]} 1, 24 -- MIME-version: 1.0 1, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 24 -- References: {200607111913.k6BJDd9P012443-at-ns.microscopy.com} 1, 24 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.7.11.121432 ==============================End of - Headers==============================
You are cordially invited to a two day, hands-on, High Pressure Freezing workshop on Aug 3 & 4, co-hosted by The Department of Molecular Genetics and Cell Biology and The Bioscience Division at The University of Chicago. The workshop will be conducted in the laboratory of Dr. Joe Austin utilizing his Bal-Tec HPM 010 High Pressure Freezer and related cryo EM sample preparation equipment. Lectures Thursday morning: 09:30 - 11:30 (Dr. Andres Kaech, Life Science Application Manager, Bal-Tec AG and Dr. Joe Austin) followed by lunch and hands-on practical lab work and instruction until 5:00pm The workshop practical sessions will continue Friday morning from 09:00 until noon Lecture room space is limited to approximately 30 people and the hands-on practical work group is restricted to a maximum of 8 participants so we encourage you to sign up early and reserve your spot. There is no charge to attend the workshop and Thursday's lunch and the Friday morning refreshments are provided with the compliments of Boeckeler Instruments.
Please email Cheryl Johnson (Cheryl-at-boeckeler.com) or Dave Roberts (dave-at-boeckeler.com) for additional details and to reserve your place.
If you are attending the Microscopy & Microanalysis meeting the University of Chicago is a 30 minute ride by cab or car from Navy Pier.
Sincerely
Dave Roberts Bal-Tec RMC Boeckeler Instruments Inc 4650 S. Butterfield Drive Tucson, AZ 85714 Tel: 520-745-0001 Fax: 520-745-0004 www.baltec-rmc.com
==============================Original Headers============================== 12, 20 -- From dave-at-boeckeler.com Tue Jul 11 17:14:34 2006 12, 20 -- Received: from txslsmtp2.vzwmail.net (txslsmtp2.vzwmail.net [66.174.85.156] (may be forged)) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6BMEYdN001717 12, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Jul 2006 17:14:34 -0500 12, 20 -- Received: from tweedledee (smtp.vzwmail.net [66.174.85.25]) 12, 20 -- (authenticated bits=0) 12, 20 -- by txslsmtp2.vzwmail.net (8.12.9/8.12.9) with ESMTP id k6BMEQgj002682 12, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Jul 2006 22:14:29 GMT 12, 20 -- Message-Id: {200607112214.k6BMEQgj002682-at-txslsmtp2.vzwmail.net} 12, 20 -- From: "Dave Roberts" {dave-at-boeckeler.com} 12, 20 -- To: {microscopy-at-msa.microscopy.com} 12, 20 -- Subject: High Pressure Freezing Workshop 12, 20 -- Date: Tue, 11 Jul 2006 15:14:22 -0700 12, 20 -- MIME-Version: 1.0 12, 20 -- Content-Type: text/plain; 12, 20 -- charset="us-ascii" 12, 20 -- Content-Transfer-Encoding: 7bit 12, 20 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 12, 20 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 12, 20 -- Thread-Index: Acak/GnWD2jcXp0qRbaI1+GPf3gr7A== ==============================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 mckee-at-helix.mgh.harvard.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mckee-at-helix.mgh.harvard.edu Name: Mary McKee
Organization: MGH
Title-Subject: [Filtered] decalcification of mouse nasal bones
Question: We need to decalcify adult mouse nasal tissue for parafin embedding and possibly also for epon embedding. Does anyone have protocols for this? Thanks in advance.
Most general suppliers of labware and consumables have RNase/DNase siliconized tubes of varying sizes, ranging from 0.2 to 2.0ml. Just call your local sales rep, or go to the molecular biology people and they will be able to give you the information. In fact, they will probably just give you the tubes, they cost less than 10¢ Cdn/tube for most makes. Some expensive tubes may run up to about 15¢ Cdn each.
Alternatively, pretty well every sterile, individually wrapped polypropylene tube with cap - 6ml, is RNase/DNase free even if they are not certified.
The other point is how you are extracting the nucleic acids. If they are using guanidinium extraction then the chaotropic salts are extremely dissociative, and I have yet to see an RNase survive the extraction process. If you put the tissue directly into the Extraction Buffer system you will not have to worry about RNase/DNase until the elution stage. And I add RNasin to that stage to prevent activity then. Check with the supplier of the extraction kit you will use, their sales reps will give you fairly good advice as to the extraction process. While I have preferences, it probably does not make much difference whether you use Invitrogen, Qiagen or Roche (I apologize to those whom I am forgetting just now) or whether it is spin column or magna-bead. They all work.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Bldg 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone: 204-789-3313 Pager: 204-931-9354 Cell: 204-781-1502 Fax: 204-789-3926
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Email: goldiyadav-at-yahoo.com Name: Vikas Yadav
Organization: JNU
Title-Subject: [Filtered] Golgi Staining: Rapid Golgi method of Stenasus
Question: I am trying to stain dendritic spines of rat brain using the rapid golgi method. Below are the details of the protocol i use:
RAPID GOLGI
1) 5mm slices of brain 2) Fixing solution (4 days) K2Cr2O7 ñ 5g Chloral hydrate- 5g Formaldehyde- 6ml Glutaraldehyde ñ 5ml DMSO- 5 drops Distilled H2O- 100 ml Change the solution every 2 days for 4 days. 3) Wash in 0.75%AgNO3(4 times) 4) Keep in 0.75% AgNO3- 4 days 5) Dehydration- 50%, 70%, 90% (2 hours each) 100% 2 changes of 1 hour each. 6) Ethyl Alcohol: Diethyl ether 1:1 ratio 1 hour 7) Embedding in celloidin (hot celloidin method) 8) Block (30% celloidin).
My problem is that while tissue fixation, embedding and sectioning seem to be OK the stain has apparently not penetrated deep into the tissue. While the cortical cells are well stained the deeper cells are not as well stained.
Secondly after 10 days the intensity of the stain seems to have gone down.
What could be the cause of this problewm? Can anyone help me?
I have so far been able to avoid having to section cells on their substrate (I usually pop them off) for TEM, but a client wants to look at the interaction of biofilms with the substrate. I'm looking for an opinion from anyone who has tried to thin section films on Thermanox, Permanox, or has cast their own resin slides (I think I've seen a mold for that). How likely is it that any of these materials will delaminate after embedding in epoxy resin? Acrylic resin? Slide shapes are preferable to coverslip types.
Any advice appreciated!
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 Wed Jul 12 14:59:40 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 k6CJxdQC032019 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Jul 2006 14:59:40 -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 k6CJxbve026319 7, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Jul 2006 09:59:38 -1000 (HST) 7, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k6CJxaEU026315 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Jul 2006 09:59:37 -1000 (HST) 7, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 7, 19 -- Date: Wed, 12 Jul 2006 09:59:36 -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: Sectioning Thermanox, Permanox, resin 7, 19 -- Message-ID: {Pine.GSO.4.21.0607120935370.26169-100000-at-halia} 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
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Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
Sorry for any delay in my reply. Overseas and having trouble sending mail!!??
Well I have kept out of the gold coated cathode debate for those that know me will already have guessed my reply!
However, I too know of the RCA gold plated cathode and 100% agree with John Bozzola in that I believe sputtering of gold would not be efficient in this case.
The answer my clients would expect is "why go to such lengths when cleaning a cathode assembly should be simple?" This is what we use -
"The cathode assembly is placed, aperture face upwards, in a beaker of stock ammonia solution diluted 3 parts ammonia to one part water. The stock solution is thought to be about 40% ammonia. After 15 minutes in the ultrasonic cleaner the beaker is placed under running water and thoroughly flushed through. Care is taken to ensure that none of the clamping or alignment screws had fallen out of the cathode assembly and could be flushed away! The cathode is then washed with alcohol before being dried with a hair drier. A new filament is fitted and centred. The assembly is checked for cleanliness by observing with a 20X lens prior to re installation in the microscope. Total time for this procedure should be less than 25 minutes break to pump down."
If the contamination in the nose of the cathode is obstinate use a little metal polish but only a polish soluble in ammonia. A follow the above procedure again but for only two or three minutes ammonia solution agitation.
The full story is available on our web site.
Steve Chapman Protrain for EM training & consultancy world wide www.emcourses.com Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967
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Email: moxleyma-at-slu.edu Name: Michael A. Moxley
Question: The correction collar on an olympus objective is very stiif almost impossible to turn. This is an inheirited scope so before investing in repairs is there anything I can do about this. Thanks
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Email: dmcdaniel-at-usuhs.mil Name: Dennis McDaniel
Organization: Uniformed Services University of the Health Sciences
I have many years experience with various types of microscopy but am relatively new to immunoEM. I am looking for a good starting protocol for gold labeling surface antigens on bacterial spore surfaces. Any nudge in the right direction would be greatly appreciated. Thanks.
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Email: m_jarnik-at-fccc.edu Name: M. Jarnik
Organization: Fox Chase Cancer Ctr
Title-Subject: [Filtered] ReAsH labeling for EM
Question: Does anybody on the list have hands-on experience with this reagent? I would like to use it both for flurescent microscopy and EM (after DAB photoconversion). A reference to a technically oriented publication would be appreciated as well.
There are many publications about labelling of suface antigens using large or small sized gold particles, pre- and postembedding.
A pre-embedding approach may be the easiest way to start, although LPS may cause steric hindrance and mask antigens in intact cells. in such cases cryoultramicrotomy may be needed to expose antigens. See e.g.
Tommassen J, Leunissen J, van Damme-Jongsten M, Overduin P. Failure of E. coli K-12 to transport PhoE-LacZ hybrid proteins out of the cytoplasm. EMBO J. 1985 Apr;4(4):1041–1047 Bosch D, Leunissen J, Verbakel J, de Jong M, van Erp H, Tommassen J. Periplasmic accumulation of truncated forms of outer-membrane PhoE protein of Escherichia coli K-12. J Mol Biol. 1986 Jun 5;189(3):449–455
Feel free to get in touch off-line for details, if you like.
Jan Leunissen
Aurion - President Research Advisor EM Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4797301 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz
==============================Original Headers============================== 8, 21 -- From leunissen-at-aurion.nl Thu Jul 13 20:49:35 2006 8, 21 -- Received: from fep03.xtra.co.nz (fep03.xtra.co.nz [210.54.141.243]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6E1nYUu032360 8, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 13 Jul 2006 20:49:35 -0500 8, 21 -- Received: from [192.168.1.50] (really [222.153.174.99]) by fep03.xtra.co.nz 8, 21 -- with ESMTP 8, 21 -- id {20060714014926.QHSR5275.fep03.xtra.co.nz-at-[192.168.1.50]} ; 8, 21 -- Fri, 14 Jul 2006 13:49:26 +1200 8, 21 -- In-Reply-To: {200607140038.k6E0cbKg010977-at-ns.microscopy.com} 8, 21 -- References: {200607140038.k6E0cbKg010977-at-ns.microscopy.com} 8, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 21 -- Content-Type: text/plain; charset=WINDOWS-1252; delsp=yes; format=flowed 8, 21 -- Message-Id: {39BF0265-60FB-4841-B07C-53A697A4DFB4-at-aurion.nl} 8, 21 -- Cc: Microscopy-at-microscopy.com 8, 21 -- From: Jan Leunissen {leunissen-at-aurion.nl} 8, 21 -- Subject: Re: [Microscopy] viaWWW: bacterial spore immunoEM 8, 21 -- Date: Fri, 14 Jul 2006 13:49:24 +1200 8, 21 -- To: dmcdaniel-at-usuhs.mil 8, 21 -- X-Mailer: Apple Mail (2.752.2) 8, 21 -- Content-Transfer-Encoding: 8bit 8, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6E1nYUu032360 ==============================End of - Headers==============================
I have no experience with LR-White. Yesterday I got a few ml of LR-white to try flat embedding in a petri-dish. I wanted to try chemical curing (with the accelerator) overnight at 4°C. Today the resin is still liquid and the plastic petri dish looks somewhat attacks by the product. Can't I use LR-White in plastic petri dishes (I used only ethanol)? I don't know how old is this resin, it was stored at 4°C.
I want to avoid curing in the oven, but if I cannot avoid it, I want the lowest temperature possible. I wondered if I couldn't polymerize the resin over the weekend at 40°C or 50°C.
Has someone a protocol using LR-White which would fit my wishes?
Regards,
Stephane
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Flat embedding LRW is always fun. The key is that, since it won't polymerize in the presence of oxygen, the molds holding sample and resin must be protected from oxygen by some means. We often use Thermonox cover slips (large rectangular ones) to cover the mold openings, because it is easy to peel them off after polymerization without the breakage of glass coverslips.. It is necessary to fill adjacent molds with LRW as "buffers" and cover them too, since the molds near the edges of the coverslips tend to lose lots of resin due to evaporation. This unit can be heat-polymerized like other resins or UV-polymerized in an ice chest or refrigerator to keep temperatures low. We often use molds from Ted Pella (cat. No. 10506, for example), for this purpose since they are strips held flat by a metal frame and exactly fit 22x60 mm Thermonox cover slips to seal the openings. We overfill the openings with resin so that there is overflow and the coverslips seal well. Other suppliers may have similar molds or others that would work as well.
Other options would be polymerizing in a sealed container filled with nitrogen or argon or some other gas that is not oxygen, or polymerizing in a vacuum chamber. My one experience with the latter resulted in the loss of nearly all the LRW in the mold as it evaporated, but it should be relatively easy to do some variation on the coverslip procedure to keep the resin from sublimely wandering off.
Hope this helps. Good luck.
Randy
Randy Tindall 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
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Friday, July 14, 2006 1:24 AM To: Tindall, Randy D.
Dear listers,
I have no experience with LR-White. Yesterday I got a few ml of LR-white to try flat embedding in a petri-dish. I wanted to try chemical curing (with the accelerator) overnight at 4°C. Today the resin is still liquid and the plastic petri dish looks somewhat attacks by the product. Can't I use LR-White in plastic petri dishes (I used only ethanol)? I don't know how old is this resin, it was stored at 4°C.
I want to avoid curing in the oven, but if I cannot avoid it, I want the lowest temperature possible. I wondered if I couldn't polymerize the resin over the weekend at 40°C or 50°C.
Has someone a protocol using LR-White which would fit my wishes?
Regards,
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
When using flat embedding molds with resins that creep, we found that if you put a flat-ended cryo pin into the resin, they don't creep. This works in an AFS chamber where the atmosphere is nitrogen. Elaine
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-- Dr. Elaine Humphrey Microscopy Specialist President, Microscopy Society of Canada (2003-2005) University of British Columbia e-mail: ech-at-interchange.ubc.ca
POSTDOCTORAL POSITION AVAILABLE IMMEDIATELY DEPARTMENT OF BIOCHEMISTRY AND BIOPHYSICS UNIVERSITY OF CALIFORNIA AT SAN FRANCISCO, CALIFORNIA, USA
A post-doctoral position is available immediately in the laboratory of Gang Ren at University of California at San Francisco (UCSF). The qualified candidate may participate in ongoing research projects of the Ren laboratory. The successful candidate should have a Ph.D. degree with highly motivation and interest in structure biology. Experiences in electron microscopy, computer processing/programming, crystallography and/or protein expression and purification will be advantaged.
A major research focus in the Ren Laboratory is to study the protein conformation changes at different biological conditions by electron cryo-microscopic techniques, such as single particle analysis, electron crystallography and electron tomography. Areas of interest include structural studies of lipoproteins, ABC transporters and other transmembrane proteins.
UCSF is a leading university that consistently defines health care worldwide by conducting advanced biomedical research, educating graduate students in life sciences and providing complex patient care. The Ren laboratory is located in the UCSF Mission Bay campus (beside San Francisco Bay), which is a new life sciences campus for teaching and research. The department is well equipped with electron cryo-microscopies, which include an FEI Polara 300kV FEG, helium stage microscope with Gatan Tridiem Imaging Energy Filter on a unique 4kx4k UltaCam lens-coupled CCD camera, and a FEI T20 microscope with 4kx4k CCD camera. Computer clusters, VitroBot and high pressure freezing are also available.
Interested candidates should send a CV including areas of expertise and research interest, publications list, and names and contact information for 3 references to Dr. Gang Ren at gren-at-msg.ucsf.edu.
Gang ‘Gary’ Ren, Ph.D. University of California, San Francisco Department of Biochemistry & Biophysics San Francisco, CA 94158 Email: gren-at-msg.ucsf.edu
==============================Original Headers============================== 6, 16 -- From gren-at-msg.ucsf.edu Fri Jul 14 11:15:26 2006 6, 16 -- Received: from msg.ucsf.edu (msg.ucsf.edu [169.230.20.46]) 6, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6EGFPCL023745 6, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 14 Jul 2006 11:15:25 -0500 6, 16 -- Received: from [128.249.148.38] (account gren-at-msg.ucsf.edu) 6, 16 -- by msg.ucsf.edu (CommuniGate Pro WebUser 4.2.8) 6, 16 -- with HTTP id 3706385 for Microscopy-at-microscopy.com; Fri, 14 Jul 2006 09:15:25 -0700 6, 16 -- From: "Gang 'Gary' Ren" {gren-at-msg.ucsf.edu} 6, 16 -- Subject: Postdoctoral Position Available at UCSF 6, 16 -- To: Microscopy-at-microscopy.com 6, 16 -- X-Mailer: CommuniGate Pro WebUser Interface v.4.2.8 6, 16 -- Date: Fri, 14 Jul 2006 09:15:25 -0700 6, 16 -- Message-ID: {web-3706385-at-msg.ucsf.edu} 6, 16 -- MIME-Version: 1.0 6, 16 -- Content-Type: text/plain; charset="ISO-8859-1"; format="flowed" 6, 16 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (cartoonish2-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, July 15, 2006 at 06:58:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both cartoonish2-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: cartoonish2-at-yahoo.com Name: nish pandya
Education: Graduate College
Location: washington dc usa
Title: career
Question: Hi.
Wanted to get your advice on how I can become a Microscopist. I have a Master's degree in biomedical engineering from the Univ of Miami. My graduate work was in biomedical optics and laser applications. I also did some research at Duke Univ. For the past 4 years, I have worked as a Software Engineer but I want to get back into the field of optics. Do you have any suggestions on how to get into the field ? I am willing to further my education. I just don't know where to look for resources. Thanks.
Question: Once again the MSA Education Committee is organizing Exhibitor Demonstrations and Tutorials at the MM2006 Meeting. These tutorials will take place on Tuesday, August 1 starting at 5:00 pm in the Navy Pier Exhibit Hall. These mini-seminars or tutorial demonstrations are held in the booths of the participating companies after the xhibit Hall is closed to non-participants.
Reservations sheets with titles and descriptions will be at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA MegaBooth soon, since the demonstrations get filled up quickly!
You may download a PDF file with an extended outline of the participating Exhibitors as well as titles of all the presentations at http://mm2006.microscopy.org . Then follow the links to the Exhibitors or Tutorials pages.
Dr. Andreas Holzenburg MSA Education Committee - Exhibitors Tutorials SubCommittee Chair
Why would you want to do that? Any idiot with a microscope can be called a microscopist. It is just like anyone with a camera can be a landscape photographer. Semi-true but much more complicated. Been there, done that.
You can make much more income doing software work than traditional tech SEM work. Frankly, I don't see EM as a big money maker or legacy venue. You are way better off where you are. And you can grow to bigger venues.
If you add computer engineering to your CV, that will IMO greatly assist you. Stop and think about what you want to do versus what you need to do. Your personal preferences may override all other inputs. That is fine. Just be aware of the consequences and cost of each decision.
But fundamentally--why do you want to be a microscopist? TEM, SEM, STEM, AFM, etc.? You will get solicitations that expect you to have a Ph.D in something and scads of experience in all of the specific units that the solicitor has. Either this is for one specific person or for a phantom.
Sigh....the EM folks may not like this posting. But it is the truth and from the heart.
Why am I a microscopist? I love it. EM or LM. But I am also an EE. The SEM and LM are merely tools. But they are not simple to use. However, adding EE is not a simple nor easy process. You will find this to be akin to computer engineering.
gary g.
At 08:05 AM 7/15/2006, you wrote:
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I did not mean to sound harsh. If I did, I apologize for that.
What I was trying to say, obviously with a poor approach, was that to do microscopy well requires a lot of training, work and perhaps a bit of luck. But after all of the training and course work, what type of job can one find? The MRS Bulletin lists a few job offerings. Most require a Ph.D. and typically pay $60K or so. Some jobs are available as "technicians" but these usually pay less. It is interesting that the majority of microscopy job openings are published in non-microscopy materials. Zero in Microscopy Today and I do not recall seeing one in an M&M issue--well, maybe one some time ago.
I don't like low paying EM jobs. That is why I'm an EE. The EM is simply a tool. But it is a great tool. And it can do really fantastic things.
You are right about optics. He made the initial venture into microscopy but it sounds like he really wants to get into optical physics. That is different.
gary g.
At 10:51 AM 7/16/2006, you wrote: } Hello Gary: } } I think you are very harsh to Nish. I am not sure you gave him/her the } proper advice sought. If you love microscopy with its low pay, someone } else could, don't you think so? } } Nish can get into microscopy (actually, optics, as he/she seems to } suggest) via a formal training in materials science/solid state } physics. Since he/she already has a degree in an engineering } discipline, he/she would naturally fit into materials/solid state } physics. What if Nish is intersted in synchrotron science? There are } several "optics" thing in that area too. } } Thanks and have a nice day. } } Ike Oguocha } --- gary-at-gaugler.com wrote: } } } Why would you want to do that? Any idiot with a } } microscope can be called a microscopist. It is just } } like anyone with a camera can be a landscape photographer. } } Semi-true but much more complicated. Been there, done that. } } } } You can make much more income doing software } } work than traditional tech SEM work. Frankly, } } I don't see EM as a big money maker or legacy } } venue. You are way better off where you are. } } And you can grow to bigger venues. } } } } If you add computer engineering to your CV, that } } will IMO greatly assist you. Stop and think about } } what you want to do versus what you need to do. } } Your personal preferences may override all other } } inputs. That is fine. Just be aware of the consequences } } and cost of each decision. } } } } But fundamentally--why do you want to be a microscopist? } } TEM, SEM, STEM, AFM, etc.? You will get solicitations that } } expect you to have a Ph.D in something and scads of } } experience in all of the specific units that the solicitor } } has. Either this is for one specific person or for a phantom. } } } } Sigh....the EM folks may not like this posting. But it is } } the truth and from the heart. } } } } Why am I a microscopist? I love it. EM or LM. But I } } am also an EE. The SEM and LM are merely tools. But they } } are not simple to use. However, adding EE is not a simple nor } } easy process. You will find this to be akin to computer } } engineering. } } } } gary g. } } } } } } At 08:05 AM 7/15/2006, you wrote: } } } } } } } } } on Saturday, July 15, 2006 at 06:58:20 } } } Remember to consider the Grade/Age of the student when considering } } } the Question } } } } } } Email: cartoonish2-at-yahoo.com } } } Name: nish pandya } } } } } } Education: Graduate College } } } } } } Location: washington dc usa } } } } } } Title: career } } } } } } Question: Hi. } } } } } } Wanted to get your advice on how I can become a Microscopist. I } } } have a Master's degree in biomedical engineering from the Univ of } } } Miami. My graduate work was in biomedical optics and laser } } } applications. I also did some research at Duke Univ. } } } For the past 4 years, I have worked as a Software Engineer but I } } } want to get back into the field of optics. } } } Do you have any suggestions on how to get into the field ? I am } } } willing to further my education. I just don't know where to look } } } for resources. } } } Thanks. } } } } } } } } } } ___________________________________________________________ } All new Yahoo! Mail "The new Interface is stunning in its simplicity } and ease of use." - PC Magazine } http://uk.docs.yahoo.com/nowyoucan.html
==============================Original Headers============================== 8, 21 -- From gary-at-gaugler.com Sun Jul 16 13:21:56 2006 8, 21 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6GILtrb015074 8, 21 -- for {microscopy-at-microscopy.com} ; Sun, 16 Jul 2006 13:21:56 -0500 8, 21 -- Received: (qmail 1770 invoked from network); 16 Jul 2006 11:21:54 -0700 8, 21 -- Received: by simscan 1.1.0 ppid: 1767, pid: 1768, t: 0.1830s 8, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 8, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 8, 21 -- by qsmtp2 with SMTP; 16 Jul 2006 11:21:54 -0700 8, 21 -- Message-Id: {7.0.1.0.2.20060716110359.02528658-at-gaugler.com} 8, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 8, 21 -- Date: Sun, 16 Jul 2006 11:21:55 -0700 8, 21 -- To: Ike Oguocha {oguocha-at-yahoo.com} 8, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 8, 21 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: how I can become a 8, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} , cartoonish2-at-yahoo.com 8, 21 -- In-Reply-To: {20060716175114.49347.qmail-at-web53715.mail.yahoo.com} 8, 21 -- References: {200607160618.k6G6ID7P015936-at-ns.microscopy.com} 8, 21 -- {20060716175114.49347.qmail-at-web53715.mail.yahoo.com} 8, 21 -- Mime-Version: 1.0 8, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Nish Pandya wrote: =========================================================== Wanted to get your advice on how I can become a Microscopist. I have a Master's degree in biomedical engineering from the Univ of Miami. My graduate work was in biomedical optics and laser applications. I also did some research at Duke Univ. For the past 4 years, I have worked as a Software Engineer but I want to get back into the field of optics. Do you have any suggestions on how to get into the field ? I am willing to further my education. I just don't know where to look for resources. Thanks. ========================================================== I am not sure there is any "one" best way that one becomes a microscopist. I am sure that we could even have a lively dialogue as to just what constitutes being a "microscopist". But there are numerous ways one could "become" a microscopist, from enrolling in courses to taking a position in a setting where you would be working under an experienced microscopist for a period of time.
But I would like to comment on previous comments to your question about opportunities in microscopy. From my vantage point, the opportunities have never appeared greater in the nearly 40 years since leaving graduate school myself. Certainly in the USA, there has been a huge proliferation of new start-up companies in areas such as nanotechnology and biomaterials, and one just does not get very far in those areas without having on board an experienced microscopy department and laboratory. There is also, again, at least in the USA, a growth in the number of junior colleges and even high schools that are putting in SEMs for the benefit of their students, and of course, one knowledgeable about microscopy is needed for those institutional situations as well. Anyone attending several recent exhibitions of the MSA (e.g. the "M&M" meeting) would also see that there is an ever increasing number of firms offering microscopy-related products, and with increasingly greater sophistication and technology, and such firms could not operate without employing an increasingly larger number of experienced microscopists.
The issue of salary is indeed important and I see our industry as being very very healthy and it would not be attracting the good people it needs to grow and innovate if it was not paying the kinds of salaries that could attract the best of minds. Certainly there is a spread in terms of what people earn, just as there is for just about any other walk of life. But I would not agree with the suggestion that the opportunities, professional or financial, are not present for those contemplating a career in microscopy.
Chuck
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==============================Original Headers============================== 9, 26 -- From cgarber-at-2spi.com Sun Jul 16 23:17:39 2006 9, 26 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 9, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6H4Hdr4023195 9, 26 -- for {microscopy-at-msa.microscopy.com} ; Sun, 16 Jul 2006 23:17:39 -0500 9, 26 -- Received: from yourb27fb1c401 (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) 9, 26 -- (authenticated bits=0) 9, 26 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id k6H4HZCi009309 9, 26 -- for {microscopy-at-msa.microscopy.com} ; Mon, 17 Jul 2006 00:17:39 -0400 9, 26 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] 9, 26 -- X-IDV-HELO: yourb27fb1c401 9, 26 -- X-IDV-Authenticated-User: cgarber 9, 26 -- Message-ID: {009e01c6a957$ef454120$6401a8c0-at-yourb27fb1c401} 9, 26 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 9, 26 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 9, 26 -- Subject: More on "becoming a microscopist" 9, 26 -- Date: Mon, 17 Jul 2006 00:17:35 -0400 9, 26 -- MIME-Version: 1.0 9, 26 -- Content-Type: text/plain; 9, 26 -- format=flowed; 9, 26 -- charset="iso-8859-1"; 9, 26 -- reply-type=original 9, 26 -- Content-Transfer-Encoding: 7bit 9, 26 -- X-Priority: 3 9, 26 -- X-MSMail-Priority: Normal 9, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 9, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
Obviously the plastic container was the cause of the bad polymerization since curing in Falcon tubes or eppendorfs works and the blocks are ready in 20 min with the accelerator. I am currently trying to find the ideal container for my purpose but it is really not easy. It must be in PE or PP, have a flat bottom and be large enough to accomodate 18x18 coverslips and it must be one-use.
Chemical curing is really easier than the other methods because one does not need to care about the presence of air and it is much (and I mean much) faster. Notwithstanding I wonder how it is possible to heat in oven AND at the same time keep a nitrogen atmosphere. Why couldn't one simply cover the resin with oil to avoid contact with air? Or another liquid which does not mixes with the resin?
BTW, can someone tell me what plastic are made the caps of 50 ml falcon tubes? I couldn't find this information.
regards,
Stéphane
--- nizets2-at-yahoo.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } I have no experience with LR-White. Yesterday I got } a } few ml of LR-white to try flat embedding in a } petri-dish. I wanted to try chemical curing (with } the } accelerator) overnight at 4°C. Today the resin is } still liquid and the plastic petri dish looks } somewhat } attacks by the product. Can't I use LR-White in } plastic petri dishes (I used only ethanol)? I don't } know how old is this resin, it was stored at 4°C. } } I want to avoid curing in the oven, but if I cannot } avoid it, I want the lowest temperature possible. I } wondered if I couldn't polymerize the resin over the } weekend at 40°C or 50°C. } } Has someone a protocol using LR-White which would } fit } my wishes? } } Regards, } } Stephane } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam } protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 7, 18 -- From nizets2-at-yahoo.com Fri Jul 14 01:22:05 } 2006 } 7, 18 -- Received: from web37414.mail.mud.yahoo.com } (web37414.mail.mud.yahoo.com [209.191.87.67]) } 7, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k6E6M5XL016952 } 7, 18 -- for {microscopy-at-microscopy.com} ; Fri, 14 } Jul 2006 01:22:05 -0500 } 7, 18 -- Received: (qmail 88984 invoked by uid } 60001); 14 Jul 2006 06:22:04 -0000 } 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; } c=nofws; } 7, 18 -- s=s1024; d=yahoo.com; } 7, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 7, 18 -- } b=iz7y9kHVNTA7vVqeF0/eP8sTXePnPx2ykJjmp547BWX3234uTXAOwrmqeVZ1PhYMSnv3A28RmFE3xzXMxuWyjQ0aoq8Ts3j87PpI2B87dOLqPrC1jyPDlEvomV5DpRbpma7L0g9GrXaMHSse319FTP3AwxvFvi0TEavGtOqyQys= } ; } 7, 18 -- Message-ID: } {20060714062204.88982.qmail-at-web37414.mail.mud.yahoo.com} } 7, 18 -- Received: from [80.122.101.102] by } web37414.mail.mud.yahoo.com via HTTP; Thu, 13 Jul } 2006 23:22:04 PDT } 7, 18 -- Date: Thu, 13 Jul 2006 23:22:04 -0700 (PDT) } 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 7, 18 -- Subject: Urgent HELP: LR-White informations } needed } 7, 18 -- To: microscopy-at-microscopy.com } 7, 18 -- MIME-Version: 1.0 } 7, 18 -- Content-Type: text/plain; } charset=iso-8859-1 } 7, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
I'm a generalist. I would not want it any other way. I use PLM, phase contrast, dispersion staining, SEM, EDS, TEM and microchemical testing. I've polished thin sections of minerals, coal and pencil urchin spines. I made blood spears and cut thin sections of wood, rubber and plastics. I've polished metal and cross sections of dipped kevlar cord. I can't imaging spending 30 years doing citric acid titrations or measuring beach sand particles.
Do I have a career? No, I have an advocation. Am I rich? No, but I've made a good living doing what I would do as a hobby if I could afford the equipment.
You want to be a microscopist? Blow off all of this advise and study microscopy. Learn about PLN and image analysis. Study SEM and TEM/EELS. Learn to take photomicrographs that people want to see. Poke around in microchemical testing and staining. Look at some minerals from the point of view as a geologist and then as the forensic microscopist. Associate with other microscopists. And when you find an area you like stay with it.
My first job out of college was a QC chemist. I hated it. I started sleeping later and later every day. I started at work arriving later and later every day. Fortunately my interest in microscopy found me a position I wanted. I like being a generalist. Yes I look forward to Friday night, but come Sunday night, I start thinking about the job and most of the time I'm fired up about starting a new work week. I know people who make a lot more money, but they also hate their job.
Now I gotta go... I have about 500 particle of carbon black i have to measure and i can't wait to find out if they'll match the earlier data!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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==============================Original Headers============================== 12, 18 -- From frank.karl-at-degussa.com Mon Jul 17 08:03:16 2006 12, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HD3FCL018106 12, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 17 Jul 2006 08:03:15 -0500 12, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 12, 18 -- by framailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k6HD3BAQ030210 12, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 17 Jul 2006 15:03:12 +0200 12, 18 -- In-Reply-To: {200607170419.k6H4JEPW025439-at-ns.microscopy.com} 12, 18 -- Subject: Re: [Microscopy] More on "becoming a microscopist" 12, 18 -- To: microscopy-at-msa.microscopy.com 12, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 12, 18 -- Message-ID: {OF7C0E96DB.EA10FE62-ON852571AE.0045E79E-852571AE.0047B1FC-at-degussa.com} 12, 18 -- From: frank.karl-at-degussa.com 12, 18 -- Date: Mon, 17 Jul 2006 09:03:06 -0400 12, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 12, 18 -- 07/17/2006 08:03:13 AM 12, 18 -- MIME-Version: 1.0 12, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Yep, flat embedding with LR-White has some problems. You are correct =20=
that the plastic dish is a problem. Oxygen is also a problem. Here =20 is what I have done.
Coat TC plates with sterile, molten 1.5% Agar, 0.5% Gelatin in =20 ddH2O. Wash around the inside of the plate and on the top of the =20 plate cover, pour out the excess and let dry. Set up cells. Cells may not adhere to this coating as well as to =20 plastic, so you may need to increase the length of cell binding =20 incubations (and be more gentle with future treatments). Do all of the things you normally do. Change the LR-White several =20 times. The final should fill the TC dish to over full. Place the =20 cover upside-down on top of the Dish (diagonally) to exclude all air.
The covering of the Agar & Gelatin will keep the LR-White from =20 interacting with the plastic of the dish. Having the dish full of LR- =20=
White and putting the cover on the dish (Agar/Gelatin side in contact =20=
with the LR-White) will exclude most of the oxygen. This has worked =20 for me in the past. Let me know if you have any questions.
David
On Jul 13, 2006, at 11:26 PM, nizets2-at-yahoo.com wrote:
--| ------ --| --| Dear listers, --| --| I have no experience with LR-White. Yesterday I got a --| few ml of LR-white to try flat embedding in a --| petri-dish. I wanted to try chemical curing (with the --| accelerator) overnight at 4=B0C. Today the resin is --| still liquid and the plastic petri dish looks somewhat --| attacks by the product. Can't I use LR-White in --| plastic petri dishes (I used only ethanol)? I don't --| know how old is this resin, it was stored at 4=B0C. --| --| I want to avoid curing in the oven, but if I cannot --| avoid it, I want the lowest temperature possible. I --| wondered if I couldn't polymerize the resin over the --| weekend at 40=B0C or 50=B0C. --| --| Has someone a protocol using LR-White which would fit --| my wishes? --| --| Regards, --| --| Stephane
==============================Original Headers============================== 13, 20 -- From Elliott-at-arizona.edu Mon Jul 17 09:19:41 2006 13, 20 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HEJfou029415 13, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 17 Jul 2006 09:19:41 -0500 13, 20 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 13, 20 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 32944EA98BB 13, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 17 Jul 2006 07:19:41 -0700 (MST) 13, 20 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 13, 20 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id E637EEA9624 13, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 17 Jul 2006 07:19:39 -0700 (MST) 13, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 13, 20 -- Content-Transfer-Encoding: 7bit 13, 20 -- Message-Id: {C46C7CD7-5FB2-423F-B14C-4A426291871E-at-arizona.edu} 13, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 13, 20 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 13, 20 -- From: David Elliott {Elliott-at-arizona.edu} 13, 20 -- Subject: Re: LR-White information needed 13, 20 -- Date: Mon, 17 Jul 2006 07:19:38 -0700 13, 20 -- X-Mailer: Apple Mail (2.752.2) 13, 20 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Hi Listers, Anyone out there have a set of operator's manuals (I have the schematics, so I won't need these) for the Stereoscan 260 SEM that I may copy or purchase? Please contact me off list, thanks.
Gary Easton
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==============================Original Headers============================== 7, 20 -- From garyeaston-at-scannerscorp.com Mon Jul 17 09:27:11 2006 7, 20 -- Received: from omr1.networksolutionsemail.com (omr1.networksolutionsemail.com [205.178.146.51]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HERAdh007174 7, 20 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 09:27:10 -0500 7, 20 -- Received: from mail.networksolutionsemail.com (ns-omr1.mgt.hosting.dc2.netsol.com [10.49.6.64]) 7, 20 -- by omr1.networksolutionsemail.com (8.13.6/8.13.6) with SMTP id k6HER8JL019862 7, 20 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 10:27:08 -0400 7, 20 -- Received: (qmail 24039 invoked by uid 78); 17 Jul 2006 14:26:48 -0000 7, 20 -- Received: from unknown (HELO ?192.168.1.46?) (70.22.76.210) 7, 20 -- by 10.49.36.64 with SMTP; 17 Jul 2006 14:26:48 -0000 7, 20 -- Received: from 127.0.0.1 (AVG SMTP 7.1.394 [268.10.1/389]); Mon, 17 Jul 2006 10:26:41 -0400 7, 20 -- Message-ID: {44BB9E21.5000108-at-scannerscorp.com} 7, 20 -- Date: Mon, 17 Jul 2006 10:26:41 -0400 7, 20 -- From: "Gary M. Easton" {garyeaston-at-scannerscorp.com} 7, 20 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 7, 20 -- To: Microscopy Society of America {microscopy-at-microscopy.com} 7, 20 -- Subject: CAMBRIDGE/LEICA/LEO S260 SEM MANUALS 7, 20 -- Mime-Version: 1.0 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- Content-Type: text/plain; charset=ISO-8859-1; 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 amyhinsley-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: amyhinsley-at-yahoo.com Name: Amy
Title-Subject: [Filtered] PLM training
Question: I'm interested in attending a course for PLM for asbestos identification and so far have only been able to find the McCrone Institute. Are there other options?
This Question was submitted to Ask-A-Microscopist by (fortier-at-slu.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, July 17, 2006 at 14:24:19 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both fortier-at-slu.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: fortier-at-slu.edu Name: Joseph Fortier
Organization: Saint Louis University
Education: Graduate College
Location: Saint Louis, MO, U.S.A.
Title: critical point drying insects
Question: In order to prepare very small (less than or equal to 5mm) insects for electron microscopy, is it necessary to chemically dry them with acetone and amyl acetate in addition to CPD, if they have already been prepared in 100% ethanol?
If the samples are currently in 100% ethanol, you can go directly into the CPD device. Passage through amyl acetate is not necessary since liquid CO2 is miscible with absolute ethanol. Simply, place 5-10 ml of abs ethanol in the CPD chamber to maintain a saturated ethanol atmosphere (thereby preventing evaporation and drying of the insects), transfer the insects into the chamber and CPD as usual. Since the specimens are so small, I assume that they will be placed into appropriate carriers, otherwise they will be lost in the exchange of CO2. I do not believe that liquid CO2 will extract DNA, but I would defer to a molecular biologist if they now otherwise.
If you need more info, please do not hesitate to ask.
JB
} } Email: fortier-at-slu.edu } Name: Joseph Fortier } } Organization: Saint Louis University } } Education: Graduate College } } Location: Saint Louis, MO, U.S.A. } } Title: critical point drying insects } } Question: In order to prepare very small (less than or equal to 5mm) } insects for electron microscopy, is it necessary to chemically dry } them with acetone and amyl acetate in addition to CPD, if they have } already been prepared in 100% ethanol? } } Also, I wonder if the above chemical damage DNA? } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 9, 12 -- From zaluzec-at-microscopy.com Mon Jul 17 15:17:17 2006 } 9, 12 -- Received: from [10.255.254.48] (msdvpn24.msd.anl.gov } [130.202.238.88]) } 9, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k6HKHFk0024720 } 9, 12 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 } 15:17:16 -0500 } 9, 12 -- Mime-Version: 1.0 } 9, 12 -- X-Sender: (Unverified) } 9, 12 -- Message-Id: {p06020402c0e1a0be0f7d-at-[10.255.254.48]} } 9, 12 -- Date: Mon, 17 Jul 2006 16:17:19 -0400 } 9, 12 -- To: microscopy-at-microscopy.com } 9, 12 -- From: fortier-at-slu.edu (by way of Ask-A-Microscopist) } 9, 12 -- Subject: AskAMicroscopist: critical point drying insects } 9, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers==============================
-- ############################################################## 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 Web: http://www.siu.edu/~image/ ##############################################################
Thomas A. Kubik & Assoc. (TAKA) P.O. Box 208 Greenlawn, NY 11704 Phone: 516-261-2117
Peter M. Cooke Microscopy Instruction, Consultation & Analysis (MICA) 5807 N. Maplewood Chicago, IL 60659 Phone: 773-334-2240 pmcooke-at-earthlink.net
I do not know much about TAKA, so I can't give you any first hand advise about the company.
I can provide you some information about MICA though. Peter was an instructor at McCrone for many years before starting up his own company. I took my PLM training courses with him at McCrone and have used his company to train new PLM analysts at two laboratories which I have managed. His courses are taught both in Chicago and on-site if needed. I consider him the best asbestos PLM trainer in the USA. (I also have NO financial interest in MICA).
Good luck in whichever training course you choose.
Steven Grevelis Asbestos Laboratory Manager Groundwater Analytical, Inc. Buzzards Bay, MA 02532 Ph. 508.759.4441 Fax 508.759.4475 www.groundwateranalytical.com
-----Original Message----- X-from: amyhinsley-at-yahoo.com [mailto:amyhinsley-at-yahoo.com] Sent: Monday, July 17, 2006 4:30 PM To: sgrevelis-at-groundwateranalytical.com
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Email: amyhinsley-at-yahoo.com Name: Amy
Title-Subject: [Filtered] PLM training
Question: I'm interested in attending a course for PLM for asbestos identification and so far have only been able to find the McCrone Institute. Are there other options?
Go from 100% ethanol directly to the CPD using ethanol as your transition solvent. The other solvents are not necessary.
I have also just recently run across a paper that uses the CPD to reverse the effects of chemical fixation with formalin allowing sequencing so I would surmise damage if any is negligible.
Good luck
Scott Whittaker NMNH Imaging Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-633-0891
-----Original Message----- X-from: fortier-at-slu.edu [mailto:fortier-at-slu.edu] Sent: Monday, July 17, 2006 4:21 PM To: Whittaker, Scott
This Question was submitted to Ask-A-Microscopist by (fortier-at-slu.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, July 17, 2006 at 14:24:19 Remember to consider the Grade/Age of the student when considering the Question ------------------------------------------------------------------------ --- Please reply to both fortier-at-slu.edu as well as to the Microscopy Listserver ------------------------------------------------------------------------ ---
Email: fortier-at-slu.edu Name: Joseph Fortier
Organization: Saint Louis University
Education: Graduate College
Location: Saint Louis, MO, U.S.A.
Title: critical point drying insects
Question: In order to prepare very small (less than or equal to 5mm) insects for electron microscopy, is it necessary to chemically dry them with acetone and amyl acetate in addition to CPD, if they have already been prepared in 100% ethanol?
Hello Joseph. I just returned from Woods Hole, Massachusetts where we did some SEM on some small ants, Aphids and spiders. An entomologist there gave us a protocol where you rinse the bugs in 70% ETOH and directly sputter coat. I am not sure whether the ETOH killed them completely. We carefully mounted them on double stick tape on stubs and sputter coated them. The structure was very nice and the preparation extremely easy. Good Luck JoAnn Buchanan Stanford University School of Medicine At 01:22 PM 7/17/2006, you wrote:
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==============================Original Headers============================== 6, 21 -- From redhair-at-stanford.edu Mon Jul 17 16:25:00 2006 6, 21 -- Received: from smtp3.stanford.edu (smtp3.Stanford.EDU [171.67.20.26]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HLOxp3011171 6, 21 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 16:24:59 -0500 6, 21 -- Received: from smtp3.stanford.edu (localhost.localdomain [127.0.0.1]) 6, 21 -- by localhost (Postfix) with SMTP id 4DEAD4CA81; 6, 21 -- Mon, 17 Jul 2006 14:24:55 -0700 (PDT) 6, 21 -- Received: from BIG-DADDY.stanford.edu (ssmith-pbdsl1.Stanford.EDU [171.66.197.50]) 6, 21 -- by smtp3.stanford.edu (Postfix) with ESMTP id E38504C839; 6, 21 -- Mon, 17 Jul 2006 14:24:28 -0700 (PDT) 6, 21 -- Message-Id: {6.2.5.6.2.20060717141736.03765af0-at-stanford.edu} 6, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 6, 21 -- Date: Mon, 17 Jul 2006 14:24:27 -0700 6, 21 -- To: fortier-at-slu.edu, "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 21 -- From: JoAnn Buchanan {redhair-at-stanford.edu} 6, 21 -- Subject: Re: [Microscopy] AskAMicroscopist: critical point drying 6, 21 -- insects 6, 21 -- In-Reply-To: {200607172022.k6HKMslc032695-at-ns.microscopy.com} 6, 21 -- References: {200607172022.k6HKMslc032695-at-ns.microscopy.com} 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Thomas A. Kubik & Assoc. (TAKA) P.O. Box 208 Greenlawn, NY 11704 Phone: 516-261-2117
Peter M. Cooke Microscopy Instruction, Consultation & Analysis (MICA) 5807 N. Maplewood Chicago, IL 60659 Phone: 773-334-2240 pmcooke-at-earthlink.net
I do not know much about TAKA, so I can't give you any first hand advise about the company.
I can provide you some information about MICA though. Peter was an instructor at McCrone for many years before starting up his own company. I took my PLM training courses with him at McCrone and have used his company to train new PLM analysts at two laboratories which I have managed. His courses are taught both in Chicago and on-site if needed. I consider him the best asbestos PLM trainer in the USA. (I also have NO financial interest in MICA).
Good luck in whichever training course you choose.
Steven Grevelis Asbestos Laboratory Manager Groundwater Analytical, Inc. Buzzards Bay, MA 02532 Ph. 508.759.4441 Fax 508.759.4475 www.groundwateranalytical.com
I know Peter as well as Thomas "Tom" A. Kubik & Assoc. (TAKA); I am biased though as I teach at the McCrone Research Institute.
Peter is a good teacher and has been an NVLAP inspector.
Tom is well versed in PLM and acceptable as well; been around for a long time.
There are a couple on the West Coast (Seattle and CA) that I know of but too indirectly now and thus will not commet on because of that.
Another option is the Environmental Institute in Atlanta/Marietta, GA. Dave Hogue (formerly of GA Tech RI) runs the place and Tom Laubenthal usually teaches the PLM asbestos course. Tom has been around for years (we met when we both worked at McCrone in Atlanta in 1987) and has been an NVLAP inspector. He knows the PLM, asbestos and is not too shabby on the regs as well as field work. Unfortunately, Tom has become a little too jaded over the years (like myself) and it tends to slip into the training. Would still recommend him though.
The Environmental Institute 1300 Williams Drive Marietta, GA 30066 (770) 427-3600 (770) 421-2484 Fax info-at-tei-atl.com
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
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==============================Original Headers============================== 23, 31 -- From ph2-at-sprynet.com Mon Jul 17 18:18:05 2006 23, 31 -- Received: from elasmtp-spurfowl.atl.sa.earthlink.net (elasmtp-spurfowl.atl.sa.earthlink.net [209.86.89.66]) 23, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HNI4AY022580 23, 31 -- for {Microscopy-at-Microscopy.Com} ; Mon, 17 Jul 2006 18:18:05 -0500 23, 31 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 23, 31 -- s=dk20050327; d=sprynet.com; 23, 31 -- b=JjKbfG34jr3nM5RJe726NY/WnBuLD3DchXSqZFkI+PwaMTkKTtrKuPbNl31VMmeT; 23, 31 -- h=Received:Reply-To:From:To:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; 23, 31 -- Received: from [4.224.249.111] (helo=yourw92p4bhlzg) 23, 31 -- by elasmtp-spurfowl.atl.sa.earthlink.net with asmtp (TLSv1:RC4-MD5:128) 23, 31 -- (Exim 4.34) 23, 31 -- id 1G2cLe-0004FK-Q5; Mon, 17 Jul 2006 19:18:02 -0400 23, 31 -- Reply-To: {ph2-at-sprynet.com} 23, 31 -- From: "Tony Havics" {ph2-at-sprynet.com} 23, 31 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} 23, 31 -- Subject: PLM Training on Asbestos 23, 31 -- Date: Mon, 17 Jul 2006 19:24:28 -0500 23, 31 -- Organization: pH2 23, 31 -- Message-ID: {000201c6aa00$8b464d00$6ff9e004-at-QEPI.local} 23, 31 -- MIME-Version: 1.0 23, 31 -- Content-Type: text/plain; 23, 31 -- charset="utf-8" 23, 31 -- X-Priority: 3 (Normal) 23, 31 -- X-MSMail-Priority: Normal 23, 31 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 23, 31 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 23, 31 -- Importance: Normal 23, 31 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f9973d681e0e4feb7e488d415b7b9318c3350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 23, 31 -- X-Originating-IP: 4.224.249.111 23, 31 -- Content-Transfer-Encoding: 8bit 23, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6HNI4AY022580 ==============================End of - Headers==============================
Sorry I did not provide the reference. It had just hit my hand a couple weeks ago but not yet been entered into my database and my assistant wasn't around to ask where it was.
Formalin Removal from Archival Tissue by Critical Point Drying Biotechniques 33:604-611 September (2002).
Scott Whittaker NMNH Imaging Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-633-0891
-----Original Message----- X-from: Jim Ehrman [mailto:jehrman-at-mta.ca] Sent: Monday, July 17, 2006 7:17 PM To: Whittaker, Scott
Hello, I have used the microwave from Pella for TEM/SEM processing. However I don't have a budget for one right now.
I was wondering if anyone has tried using the new variable wattage microwaves available for consumer use? One I was considering testing is from Panasonic. The technology is explained here:
or a link if wrapping beyond the margins doesn't work: http://makeashorterlink.com/?M24131D6D
Granted, I may have to make use of water baths to avoid hot spots, but it is a far cheaper route than the $15k. The wattage for histology microwaves are variable from 20-1000 watts, or 250-750 watts. The full power of the panasonic one is 1250 watts, but has low; medium; and high wattage settings.
Anyone given these a try, or know of other commercial microwaves with variable wattage settings that report actual wattage numbers? Thanks for your help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 8, 24 -- From gvrdolja-at-nature.berkeley.edu Tue Jul 18 11:35:02 2006 8, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6IGZ1eV020273 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 11:35:02 -0500 8, 24 -- Received: from localhost (localhost [127.0.0.1]) 8, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 13872C1E69 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 09:35:00 -0700 (PDT) 8, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 8, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 8, 24 -- with ESMTP id 26824-02 for {microscopy-at-microscopy.com} ; 8, 24 -- Tue, 18 Jul 2006 09:34:45 -0700 (PDT) 8, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 8, 24 -- id 186B2C1E59; Tue, 18 Jul 2006 09:34:45 -0700 (PDT) 8, 24 -- Received: from localhost (localhost [127.0.0.1]) 8, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 0EBD3C1E48 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 09:34:44 -0700 (PDT) 8, 24 -- Date: Tue, 18 Jul 2006 09:34:44 -0700 (PDT) 8, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 8, 24 -- To: microscopy-at-microscopy.com 8, 24 -- Subject: use of microwaves for sample preparation 8, 24 -- Message-ID: {Pine.SOC.4.64.0607180904180.9949-at-nature.Berkeley.EDU} 8, 24 -- MIME-Version: 1.0 8, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
The following position is available at Virginia Commonwealth University School of Medicine.
Microscopy Technician (Position # 55122)
A position is available in the Microscopy Facility of the Department of Anatomy and Neurobiology in the School of Medicine at Virginia Commonwealth University. The facility houses confocal, multi-photon, fluorescence, and electron microscopes (TEM & SEM). The successful candidate will assist with microscopy studies of various biological systems. Duties include instructing and assisting users of the facility, sample preparation, image analysis and routine equipment maintenance.
Applicants should have excellent communication and organizational skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload. Qualifications include a degree in Biology/Life Sciences, a minimum of 2 years experience with laser scanning microscopy (confocal or multi-photon), sample preparation, digital imaging, and image analysis. Experience with electron microscopy is an asset. Computer skills are essential.
Applications are to be submitted online via the VCU Jobs website at: www.vcujobs.com
Click on the "Search Postings" link and under the "Working Title" field, select "Microscopy Technician".
--------------------------
Scott Henderson, Ph.D. Director of Microscopy Associate Professor Dept. Anatomy & Neurobiology Virginia Commonwealth University School of Medicine Sanger Hall, Rm. 9-069d 1101 East Marshall St. Richmond, VA 23298-0709
==============================Original Headers============================== 9, 28 -- From schenderson-at-vcu.edu Tue Jul 18 15:51:30 2006 9, 28 -- Received: from cedar.vcu.edu (cedar.vcu.edu [128.172.8.172]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6IKpUJv005000 9, 28 -- for {Microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 15:51:30 -0500 9, 28 -- Received: from Henderson ([128.172.29.122]) 9, 28 -- by cedar.vcu.edu (Lotus Domino Release 7.0.1FP1) 9, 28 -- with ESMTP id 2006071816512448-1271643 ; 9, 28 -- Tue, 18 Jul 2006 16:51:24 -0400 9, 28 -- Reply-To: {schenderson-at-vcu.edu} 9, 28 -- From: "Scott Henderson" {schenderson-at-vcu.edu} 9, 28 -- To: {Microscopy-at-microscopy.com} 9, 28 -- Subject: microscopy position available 9, 28 -- Date: Tue, 18 Jul 2006 16:51:23 -0400 9, 28 -- Organization: Virginia Commonwealth Unversity 9, 28 -- Message-ID: {002401c6aaab$f233d310$d361850a-at-Henderson} 9, 28 -- MIME-Version: 1.0 9, 28 -- X-Mailer: Microsoft Office Outlook 11 9, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 9, 28 -- Thread-Index: Acaqq+22V4y7UFr4SdO7xNFlc5sNMA== 9, 28 -- X-MIMETrack: Itemize by SMTP Server on CEDAR/VCU(Release 7.0.1FP1|April 17, 2006) at 07/18/2006 9, 28 -- 04:51:24 PM, 9, 28 -- Serialize by Router on CEDAR/VCU(Release 7.0.1FP1|April 17, 2006) at 07/18/2006 9, 28 -- 04:51:30 PM, 9, 28 -- Serialize complete at 07/18/2006 04:51:30 PM 9, 28 -- Content-Type: text/plain; 9, 28 -- charset="iso-8859-1" 9, 28 -- Content-Transfer-Encoding: 8bit 9, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6IKpUJv005000 ==============================End of - Headers==============================
I have a customer who would like to image in an SEM the crystal structure of PEEK (proprietary fluorocarbon polymer).
They can do it by light microscopy but the extra depth of focus in the SEM would be useful plus the ability to potentially ID inclusions by EDS.
Any suggestions? -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
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==============================Original Headers============================== 4, 14 -- From larry-at-cymru.freewire.co.uk Tue Jul 18 16:07:18 2006 4, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 4, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6IL7I08015287 4, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 18 Jul 2006 16:07:18 -0500 4, 14 -- Received: from [217.154.253.6] (th6dc-217-154-253-6.dial.mistral.co.uk [217.154.253.6]) 4, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k6IL7FDY021168 4, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 18 Jul 2006 22:07:16 +0100 4, 14 -- Mime-Version: 1.0 4, 14 -- Message-Id: {p06210200c0e2fcf1179a-at-[217.154.254.49]} 4, 14 -- Date: Tue, 18 Jul 2006 22:06:03 +0100 4, 14 -- To: Microscopy-at-MSA.Microscopy.Com 4, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 4, 14 -- Subject: Polymer Crystals by SEM? 4, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Does anyone know of an alternative to the Haskris R050 air cooled chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at least every year. The pump/motor unit will seize. Then the compressor control circuitry will fry.
Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I don't see any other maker that can match this.
If they are there, please advise.
gary g.
==============================Original Headers============================== 5, 17 -- From gary-at-gaugler.com Tue Jul 18 20:14:25 2006 5, 17 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6J1EOxO029041 5, 17 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 20:14:25 -0500 5, 17 -- Received: (qmail 30280 invoked from network); 18 Jul 2006 18:14:24 -0700 5, 17 -- Received: by simscan 1.1.0 ppid: 30277, pid: 30278, t: 0.0991s 5, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 5, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 5, 17 -- by qsmtp3 with SMTP; 18 Jul 2006 18:14:24 -0700 5, 17 -- Message-Id: {7.0.1.0.2.20060718181319.02462e60-at-gaugler.com} 5, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 5, 17 -- Date: Tue, 18 Jul 2006 18:14:25 -0700 5, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 5, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 5, 17 -- Subject: 5, 17 -- Mime-Version: 1.0 5, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Questions: Does anyone know of any companies that sell anti-BrdU antibodies that bind without the need for DNA denaturation? If not, is there an acid-free method for labelling cells in S-phase? And does anyone have a detergent-free method for labelling cells in M and S phases?
The details of this problem are as follows.
Details: I'm trying to determine is a protein toxin (directly labelled using a Molecular Probes Oregon Green 488 labelling kit) is internalised by a eukaryotic cell with respect to cell cycle. For S phase labelling I use BrdU incorporation plus a mouse monoclonal anti-BrdU from Zymed/Invitrogen. For M phase, I use murine anti-Phospho Histone H3, Ser10 (anti-PHH3). While labelling for cells in M/S phases has been successful, I have encountered a major problem with each labelling and I think they *may* be related.
Protocol: In short, non-synchronous populations of live cells are incubated with Oregon Green 488-labelled Stx (Stx-OG488) for 30min (and BrdU if labelling for cells in S phase), formalin fixed and permeabilised with 0.1% Triton X-100. For BrdU antigen retrieval, DNA is denatured for 30mins with 2M HCl and neutralised with Borax buffer. There after, M and S phase labellings are identical, in which the cells are blocked with PBS containing 20% FCS and 0.1% Triton. Primary antibodies (anti-BrdU and anti-PHH3) and secondaries conjugated to Alexa Fluor 594 are diluted in the blocking buffer and incubated for 1hr.
Hence, using dual fluorescence (green/red), we should be able to determine if cells in either M or S phase have a propensity to internalise Stx-OG488.
Here's the problem: Although visualising the cell phase is fine in the red field (texas red filter), I've found that the Stx-OG488 signal is a rather dull yellow or altogether missing when I switch to the green field (FITC filter).
Possible causes: We think this quenching is due to the use of acid denaturation for BrdU detection. I've tried 0.4M NaOH but this is highly alkaline and, as expected, also caused quenching of Stx-OG488. Also, treatment with DNase I -at- 10 U/ml for 1hr hasn't provided decent BrdU staining.
Furthermore, the continual presence of Triton X-100 during immunodetection for both M and S phases may cause dissociation of the toxin from the cell. I've tried lowering the Triton concentration and tried other detergents such as tween-20 and saponin with no improvement of the Stx-OG488 signal.
My questions are as follows: Are there any companies that sell anti-BrdU antibodies that bind without the need for DNA denaturation? If not, is there an acid-free method for labelling cells in S-phase? And does anyone have a detergent-free method for labelling cells in M and S phases?
I would be greatful for any suggestions/advice!
Thanks in advance, Damien Chong
Molecular Life Sciences Building Gate 8, Victoria Drive The University of Adelaide South Australia 5005
==============================Original Headers============================== 14, 29 -- From damien.chong-at-adelaide.edu.au Tue Jul 18 23:33:22 2006 14, 29 -- Received: from tosh.services.adelaide.edu.au (mailguard-send.adelaide.edu.au [192.43.227.21]) 14, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6J4XLsW009570 14, 29 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 23:33:22 -0500 14, 29 -- Received: from stimpy.services.adelaide.edu.au ([10.0.10.10]) 14, 29 -- by tosh.services.adelaide.edu.au with ESMTP; 19 Jul 2006 14:03:19 +0930 14, 29 -- X-IronPort-Anti-Spam-Filtered: true 14, 29 -- X-IronPort-Anti-Spam-Result: AQAAAAdTvUQN 14, 29 -- X-IronPort-AV: i="4.06,257,1149431400"; 14, 29 -- d="scan'208"; a="7422374:sNHT71535387" 14, 29 -- Received: from stimpy.services.adelaide.edu.au (localhost.localdomain [127.0.0.1]) 14, 29 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1) with ESMTP id k6J4XJpE012077 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- Received: (from apache-at-localhost) 14, 29 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1/Submit) id k6J4XJcT012068 14, 29 -- for microscopy-at-microscopy.com; Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- Received: from molb00325.molecularbiosciences.adelaide.edu.au (molb00325.molecularbiosciences.adelaide.edu.au [129.127.106.33]) 14, 29 -- by webmail.adelaide.edu.au (IMP) with HTTP 14, 29 -- for {dchong01-at-localhost} ; Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- Message-ID: {1153283599.44bdb60f20c60-at-webmail.adelaide.edu.au} 14, 29 -- Date: Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- From: Damien Chong {damien.chong-at-adelaide.edu.au} 14, 29 -- To: Microscopy ListServer {microscopy-at-microscopy.com} 14, 29 -- Subject: LM (IF): Quenching(?) with mitosis and S phase labelling 14, 29 -- MIME-Version: 1.0 14, 29 -- Content-Type: text/plain; charset=ISO-8859-1 14, 29 -- Content-Transfer-Encoding: 8bit 14, 29 -- User-Agent: Internet Messaging Program (IMP) 3.2.6 14, 29 -- X-Originating-IP: 129.127.106.33 ==============================End of - Headers==============================
I think that the antibody cannot access native DNA (and so incorporated BrU) because it is covered by proteins, so your problem does not come from the antibody (Anti-BrdU antibody are very good, at least the one from Boehringer I tried was excellent). You have to denature the DNA in order to give access to the DNA. For the permeabilization and detergent problem, you can try methanol fixation at -20°C, which permeabilizes the cells at the same time. For the detection, don't use Triton, but use serum or gelatin or fat-free milk powder to minimize non-specific interactions. If your secondary antibodies are from goat, use goat serum, not bovine serum!
Good luck,
Stephane
--- damien.chong-at-adelaide.edu.au 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 } ---------------------------------------------------------------------------- } } Send To: Microscopy-at-Microscopy.Com } } Hi Listers, } } Questions: Does anyone know of any companies that } sell anti-BrdU antibodies } that bind without the need for DNA denaturation? If } not, is there an acid-free } method for labelling cells in S-phase? And does } anyone have a detergent-free } method for labelling cells in M and S phases? } } The details of this problem are as follows. } } Details: } I'm trying to determine is a protein toxin (directly } labelled using a Molecular } Probes Oregon Green 488 labelling kit) is } internalised by a eukaryotic cell } with respect to cell cycle. For S phase labelling I } use BrdU incorporation } plus a mouse monoclonal anti-BrdU from } Zymed/Invitrogen. For M phase, I use } murine anti-Phospho Histone H3, Ser10 (anti-PHH3). } While } labelling for cells in M/S phases has been } successful, I have encountered a } major problem with each labelling and I think they } *may* be related. } } Protocol: } In short, non-synchronous populations of live cells } are incubated with Oregon } Green 488-labelled Stx (Stx-OG488) for 30min (and } BrdU if labelling for cells } in S phase), formalin fixed and permeabilised with } 0.1% Triton X-100. For BrdU } antigen retrieval, DNA is denatured for 30mins with } 2M HCl and neutralised with } Borax buffer. There after, M and S phase labellings } are identical, in which } the cells are blocked with PBS containing 20% FCS } and 0.1% Triton. Primary } antibodies (anti-BrdU and anti-PHH3) and secondaries } conjugated to Alexa Fluor } 594 are diluted in the blocking buffer and incubated } for 1hr. } } Hence, using dual fluorescence (green/red), we } should be able to determine if } cells in either M or S phase have a propensity to } internalise Stx-OG488. } } Here's the problem: } Although visualising the cell phase is fine in the } red field (texas red } filter), I've found that the Stx-OG488 signal is a } rather dull yellow or } altogether missing when I switch to the green field } (FITC filter). } } Possible causes: } We think this quenching is due to the use of acid } denaturation for BrdU } detection. I've tried 0.4M NaOH but this is highly } alkaline and, as expected, } also caused quenching of Stx-OG488. Also, treatment } with DNase I -at- 10 U/ml for } 1hr hasn't provided decent BrdU staining. } } Furthermore, the continual presence of Triton X-100 } during immunodetection for } both M and S phases may cause dissociation of the } toxin from the cell. I've } tried lowering the Triton concentration and tried } other detergents such as } tween-20 and saponin with no improvement of the } Stx-OG488 signal. } } My questions are as follows: } Are there any companies that sell anti-BrdU } antibodies that bind without the } need for DNA denaturation? If not, is there an } acid-free method for labelling } cells in S-phase? And does anyone have a } detergent-free method for labelling } cells in M and S phases? } } I would be greatful for any suggestions/advice! } } Thanks in advance, } Damien Chong } } Molecular Life Sciences Building } Gate 8, Victoria Drive } The University of Adelaide } South Australia 5005 } } ==============================Original } Headers============================== } 14, 29 -- From damien.chong-at-adelaide.edu.au Tue Jul } 18 23:33:22 2006 } 14, 29 -- Received: from } tosh.services.adelaide.edu.au } (mailguard-send.adelaide.edu.au [192.43.227.21]) } 14, 29 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k6J4XLsW009570 } 14, 29 -- for {microscopy-at-microscopy.com} ; Tue, 18 } Jul 2006 23:33:22 -0500 } 14, 29 -- Received: from } stimpy.services.adelaide.edu.au ([10.0.10.10]) } 14, 29 -- by tosh.services.adelaide.edu.au with } ESMTP; 19 Jul 2006 14:03:19 +0930 } 14, 29 -- X-IronPort-Anti-Spam-Filtered: true } 14, 29 -- X-IronPort-Anti-Spam-Result: AQAAAAdTvUQN } 14, 29 -- X-IronPort-AV: i="4.06,257,1149431400"; } 14, 29 -- d="scan'208"; a="7422374:sNHT71535387" } 14, 29 -- Received: from } stimpy.services.adelaide.edu.au } (localhost.localdomain [127.0.0.1]) } 14, 29 -- by stimpy.services.adelaide.edu.au } (8.13.1/8.13.1) with ESMTP id k6J4XJpE012077 } 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 19 } Jul 2006 14:03:19 +0930 } 14, 29 -- Received: (from apache-at-localhost) } 14, 29 -- by stimpy.services.adelaide.edu.au } (8.13.1/8.13.1/Submit) id k6J4XJcT012068 } 14, 29 -- for microscopy-at-microscopy.com; Wed, 19 } Jul 2006 14:03:19 +0930 } 14, 29 -- Received: from } molb00325.molecularbiosciences.adelaide.edu.au } (molb00325.molecularbiosciences.adelaide.edu.au } [129.127.106.33]) } 14, 29 -- by webmail.adelaide.edu.au (IMP) with } HTTP } 14, 29 -- for {dchong01-at-localhost} ; Wed, 19 Jul } 2006 14:03:19 +0930 } 14, 29 -- Message-ID: } {1153283599.44bdb60f20c60-at-webmail.adelaide.edu.au} } 14, 29 -- Date: Wed, 19 Jul 2006 14:03:19 +0930 } 14, 29 -- From: Damien Chong } {damien.chong-at-adelaide.edu.au} } 14, 29 -- To: Microscopy ListServer } {microscopy-at-microscopy.com} } 14, 29 -- Subject: LM (IF): Quenching(?) with } mitosis and S phase labelling } 14, 29 -- MIME-Version: 1.0 } 14, 29 -- Content-Type: text/plain; } charset=ISO-8859-1 } 14, 29 -- Content-Transfer-Encoding: 8bit } 14, 29 -- User-Agent: Internet Messaging Program } (IMP) 3.2.6 } 14, 29 -- X-Originating-IP: 129.127.106.33 } ==============================End of - } Headers============================== }
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This Question was submitted to Ask-A-Microscopist by (lgauthier-at-victrex.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 19, 2006 at 04:07:13 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lgauthier-at-victrex.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Dear Sir or Madam, I would like to know if it is possible to observe with a SEM the morphology (spherolites, cristals...) of a polymer (for my current study it is a PEEK). If yes in which conditions (HV or LV, pressure etc..) I should work? Thank you very much in advance.
Hello everybody, Recently I received samples that I processed for SEM (oysters). When I finished the last step of processing sputter coating I saw that my samples where not coated but the stubs were. I had a good vacuum (around 10-1 mbar), tank full of argon, current 10 mA, and I coated 6 min total. Would any body have any idea what might be the problem? It's needless to say that scoping is impossible due to charging. Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Wed Jul 19 09:24:18 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JEOHBT016664 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 09:24:18 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1G3CyE-0004oK-01 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 11:24:14 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 19 Jul 06 11:24:14 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 19 Jul 06 11:23:02 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Wed, 19 Jul 2006 11:20:29 -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: Sputter coating problems 1, 22 -- Message-ID: {44BE157D.12915.774EA9-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
I expect that you will be disappointed. Typically, elucidation of the morphology of crystalline morphology of polymers generally requires etching or staining.
Regards,
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
lgauthier-at-vict rex.com To gary.m.brown-at-exxonmobil.com 07/19/06 06:39 cc AM Subject [Microscopy] AskAMicroscopist: Please respond Observation of polymers morphology to lgauthier-at-vict rex.com
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This Question was submitted to Ask-A-Microscopist by (lgauthier-at-victrex.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 19, 2006 at 04:07:13 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lgauthier-at-victrex.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Dear Sir or Madam, I would like to know if it is possible to observe with a SEM the morphology (spherolites, cristals...) of a polymer (for my current study it is a PEEK). If yes in which conditions (HV or LV, pressure etc..) I should work? Thank you very much in advance.
We have an old Lab-line/Hooker Plant Microtome, cat #1225 with a fried motor. Lab-line is no more, and the current company (Barnstead, I think) no longer has parts for these. Does anyone have any spares or a motor or know of a source. We've been checking the used lab equipment people to no avail. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Wed Jul 19 10:15:53 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JFFqwF005507 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 10:15:52 -0500 3, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k6JFnNfd021702 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 11:49:28 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Wed, 19 Jul 2006 11:15:50 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f0623090ac0e3fca33212-at-[141.209.160.132]} 3, 20 -- Date: Wed, 19 Jul 2006 11:15:50 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: Hooker plant microtome parts 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 19 Jul 2006 15:15:50.0637 (UTC) FILETIME=[383F49D0:01C6AB46] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -3.2 () L_EXCH_MF,PORN_RP_HOOKERS_ 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
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==============================Original Headers============================== 8, 32 -- From ph2-at-sprynet.com Wed Jul 19 10:55:28 2006 8, 32 -- Received: from elasmtp-dupuy.atl.sa.earthlink.net (elasmtp-dupuy.atl.sa.earthlink.net [209.86.89.62]) 8, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JFtSSM016463 8, 32 -- for {Microscopy-at-Microscopy.Com} ; Wed, 19 Jul 2006 10:55:28 -0500 8, 32 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 32 -- s=dk20050327; d=sprynet.com; 8, 32 -- b=GCYsn7xBN4riSOfErUTzEG0LWF7ZtPZWwoEOFuTi4mvWcPWLv0NeHlQ9Apqs0M4U; 8, 32 -- h=Received:Reply-To:From:To:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; 8, 32 -- Received: from [4.224.201.228] (helo=yourw92p4bhlzg) 8, 32 -- by elasmtp-dupuy.atl.sa.earthlink.net with asmtp (TLSv1:RC4-MD5:128) 8, 32 -- (Exim 4.34) 8, 32 -- id 1G3EOU-0006Id-IN 8, 32 -- for Microscopy-at-Microscopy.Com; Wed, 19 Jul 2006 11:55:27 -0400 8, 32 -- Reply-To: {ph2-at-sprynet.com} 8, 32 -- From: "Tony Havics" {ph2-at-sprynet.com} 8, 32 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} 8, 32 -- Subject: Observation of polymers morphology 8, 32 -- Date: Wed, 19 Jul 2006 12:02:03 -0500 8, 32 -- Organization: pH2 8, 32 -- Message-ID: {002a01c6ab55$10be61c0$2f12e104-at-QEPI.local} 8, 32 -- MIME-Version: 1.0 8, 32 -- Content-Type: text/plain; 8, 32 -- charset="utf-8" 8, 32 -- X-Priority: 3 (Normal) 8, 32 -- X-MSMail-Priority: Normal 8, 32 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 8, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 8, 32 -- Importance: Normal 8, 32 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f99a782020437a081fb312cc71061d26e0350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 8, 32 -- X-Originating-IP: 4.224.201.228 8, 32 -- Content-Transfer-Encoding: 8bit 8, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JFtSSM016463 ==============================End of - Headers==============================
I have had this occur (with various specimen materials). I have not investigated it, but my first guess is significant off-gassing of the sample. Depending on the gauging configuration/location and vacuum pump capacity, it is entirely possible to indicate a good vacuum, but this may not be true in the local area of the specimen. Try pumping for quite a while* (best to slightly warm the specimen) to outgas - then try to coat.
*- I have an untrapped rough pump (only) coater, so I leak a low partial pressure of argon while long term pumping to minimize pump oil contamination.
Woody White BWXT Services
-----Original Message----- X-from: wadowska-at-upei.ca [mailto:wadowska-at-upei.ca] Sent: Wednesday, July 19, 2006 10:25 AM To: White, Woody N.
Hello everybody, Recently I received samples that I processed for SEM (oysters). When I finished the last step of processing sputter coating I saw that my samples where not coated but the stubs were. I had a good vacuum (around 10-1 mbar), tank full of argon, current 10 mA, and I coated 6 min total. Would any body have any idea what might be the problem? It's needless to say that scoping is impossible due to charging. Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Wed Jul 19 09:24:18 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JEOHBT016664 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 09:24:18 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1G3CyE-0004oK-01 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 11:24:14 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 19 Jul 06 11:24:14 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 19 Jul 06 11:23:02 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Wed, 19 Jul 2006 11:20:29 -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: Sputter coating problems 1, 22 -- Message-ID: {44BE157D.12915.774EA9-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
==============================Original Headers============================== 13, 26 -- From nwwhite-at-bwxt.com Wed Jul 19 13:00:39 2006 13, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 13, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JI0cdx028557 13, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 13:00:39 -0500 13, 26 -- Received: from ([131.184.13.224]) 13, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.1733907; 13, 26 -- Wed, 19 Jul 2006 14:00:29 -0400 13, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 13, 26 -- Wed, 19 Jul 2006 14:00:28 -0400 13, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 26 -- Content-class: urn:content-classes:message 13, 26 -- MIME-Version: 1.0 13, 26 -- Content-Type: text/plain; 13, 26 -- charset="US-ASCII" 13, 26 -- Subject: RE: [Microscopy] Sputter coating problems 13, 26 -- Date: Wed, 19 Jul 2006 14:00:28 -0400 13, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703CFFC-at-BWXSPO01.BWXS.BWXTECH.NET} 13, 26 -- X-MS-Has-Attach: 13, 26 -- X-MS-TNEF-Correlator: 13, 26 -- Thread-Topic: [Microscopy] Sputter coating problems 13, 26 -- thread-index: AcarPzmGxRaSPq8CQJGntw3be2JC/wAHMyQg 13, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 13, 26 -- To: {wadowska-at-upei.ca} , "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 13, 26 -- X-OriginalArrivalTime: 19 Jul 2006 18:00:28.0839 (UTC) FILETIME=[381D4B70:01C6AB5D] 13, 26 -- Content-Transfer-Encoding: 8bit 13, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JI0cdx028557 ==============================End of - Headers==============================
Sorry I misspelled your name before. A senior moment, I suppose.
There is a variety of tools that may be used to study the crystalline morphology of polymers. Confocal microscopy, as suggested by Tony, may be use. The choice of tool will depend on several factors including the form of the polymer (reactor particles, molded parts, blown/cast films, etc.), the scale of the structures to be analyzed (spherulites, crystallites/lamellae), the tools available for sample preparation, and the type of SEM you use (conventional, variable pressure, or field emission SEM).
Suggest you check out the latest edition of Sawyer and Grubbs Polymer Morphology. This book is a great place to start and should have a place on every lab's book shelf.
Best of luck,
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 ----- Forwarded by Gary M Brown/Baytown/ExxonMobil on 07/19/06 01:31 PM -----
gary.m.brown-at-e xxonmobil.com To gary.m.brown-at-exxonmobil.com 07/19/06 10:09 cc AM Subject [Microscopy] Re: AskAMicroscopist: Please respond Observation of polymers morphology to gary.m.brown-at-e xxonmobil.com
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Igauthier,
I expect that you will be disappointed. Typically, elucidation of the morphology of crystalline morphology of polymers generally requires etching or staining.
Regards,
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
lgauthier-at-vict rex.com To gary.m.brown-at-exxonmobil.com 07/19/06 06:39 cc AM Subject [Microscopy] AskAMicroscopist: Please respond Observation of polymers morphology to lgauthier-at-vict rex.com
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
This Question was submitted to Ask-A-Microscopist by (lgauthier-at-victrex.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 19, 2006 at 04:07:13 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lgauthier-at-victrex.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Dear Sir or Madam, I would like to know if it is possible to observe with a SEM the morphology (spherolites, cristals...) of a polymer (for my current study it is a PEEK). If yes in which conditions (HV or LV, pressure etc..) I should work? Thank you very much in advance.
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 smythen-at-musc.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: smythen-at-musc.edu Name: Nancy Smythe
Organization: Medical University of South Carolina
Title-Subject: [Filtered] help finding a book
Question: I am looking for a book that is out of print. The name is "Ultrastructural Atlas of the Inner Ear" Edited by Friedmann and Ballantyne. I had a copy but like things in labs do it grew legs and walked! Thanks
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 shaenon-at-hotmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] LKB 8800A-NM Ultrotome III Problem
Question: Hi, I am having a problem with my ultramicrotome. The specimen arm appears to be stuck in the "down" position. I cannot get it to return to the original position. I can gently push the arm up with my hand but it immediately falls back down when I let go. Does anyone have any suggestions on how I may mix this?
PS - I have tried switched back and forth between manual and automatic modes, locked the arm and returning it to the "free" position.
Thanks for the on- and off-list replies. Very helpful as usual.
An additive is definitely needed. Haskris says no--just use distilled water. that does not work for long. I use 10% mix of Ethylene Glycol (1/2G to 4.5G distilled water). That works fine. I change the nylon and toilet paper filters about once a year and change out the water mix. It is also good practice to periodically check the nylon filter for signs of algae.
The R050 chillers for Zeiss/LEO are built with quite a few options. One of these is option G. This is a hot gas bypass system. The net effect of this is that the compressor runs continuously at the factory set temperature (65F). The temperature can be changed but the compressor will still run continuously. When Zeiss installed the system, they essentially bypassed this option. They changed the thermostat set point to 72F. Doing so causes the compressor to cycle. However, by using the turbine pump which is always running, compressor on/off is not seen at the SEM. The rationale for doing this change was to bring the chilled water closer to ambient temperature of the SEM to avoid condensation.
Zeiss uses a cooling design for its electronics that IMO is much more sophisticated than others. The consequence of this is that the chiller needs to have a larger reservoir (5G) and higher flow rate. but with pumps and chiller in separate rooms from the SEM, the heat output from the SEM is very low.
All feedback about Haskris has been hugely positive. My experiences with Haskris people is also very positive. So I will stick with them.
My problem turns out to be a blown compressor rather than a seized pump. The pump motor will not operate since the high temp sensor disables the motor. When the water cools down a little from the limit, the motor runs OK. The compressor is the problem. The windings are open and the current relay is basically vaporized. So the whole compressor has to be replaced. Has anyone seen this problem before? The only known rationale for this is low line voltage. In my case, the chiller is on a Liebert Nfinity 8KVA UPS. Since I always run in inverter mode, the chiller will always get perfect power. The UPS will take out any line sags or spikes.
The concept of a single point of failure is in operation....sigh.
gary g.
At 08:30 AM 7/19/2006, you wrote:
} Gary, I have a LEO/Zeiss SEM and also the same chiller. My previous } experience } with Haskriss chillers is that some sort of water additive must be } used to eliminate } bacteria growth and corrosion. When our 1550VP was first installed } we had problems } with clogging filters and thermal alarms on the SEM until we started } treating the water. } I have been using the same additive that our building chilled water } system engineers } use at about 4 ounces per tank. I have absolutely no idea what is in } it, but we no longer } have filter issues and the Haskriss runs continuously without any } sign of corrosion } anywhere. } } } } } } Does anyone know of an alternative to the Haskris R050 air cooled } chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at } least every year. The pump/motor unit will seize. Then the } compressor control circuitry will fry. } } Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I } don't see any other maker that can match this. } } If they are there, please advise. } } gary g.
1. The cord or wire that raises and lowers the arm may be broken, slipped or stretched. Best way to determine this is to remove the front plate covering over the arm and see if the cord is intact. When you turn the manual knob, does anything move? Look specifically for the motor where the cord ends. You should see the motor turn as you move the knob. If the motor does not move, go to step 2.
2. The toothed belt that goes from the manual control knob to a small potentiometer (that raises and lowers the arm) may be worn out. Over time, this belt rots and just disintegrates. You may be moving the large manual knob but the movement is not being transferred to the potentiometer. Best way to determine this is to remove the top (3-sided, dark blue) cover from the control unit. Look for the belt that connects to the potentiometer from the manual knob. If broken, it needs to be replaced. Good luck finding one as I think they are no longer available. However, you may be able to get a similar one from a machine supply house. Check back with this listserver, as well, since someone may have one to spare. Be careful when removing covers to AVOID GETTING ELECTRICAL SHOCKS. Observe but don't probe around amongst the electronics.
3. Lastly, the potentiometer itself may be defective (if the belt is in place and moving the potentiometer control and nothing is happening with specimen arm). Another possibility: sometimes the potentiometers seize up and the belt gets stripped when it tries to turn it. Do you have any undue resistance when you move the manual control on the front of the control unit?
Try these diagnostics, take an aspirin and call me back in the morning if you have persistent problems. We are glad to help.
JB
} } Title-Subject: [Filtered] LKB 8800A-NM Ultrotome III Problem } } Question: Hi, I am having a problem with my ultramicrotome. The } specimen arm appears to be stuck in the "down" position. I cannot } get it to return to the original position. I can gently push the arm } up with my hand but it immediately falls back down when I let go. } Does anyone have any suggestions on how I may mix this? } } PS - I have tried switched back and forth between manual and } automatic modes, locked the arm and returning it to the "free" } position. } } Thanks, } Shannon } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 8, 12 -- From zaluzec-at-microscopy.com Wed Jul 19 14:44:43 2006 } 8, 12 -- Received: from [10.255.254.48] (msdvpn26.msd.anl.gov } [130.202.238.90]) } 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k6JJiffv020179 } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 } 14:44:42 -0500 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: (Unverified) } 8, 12 -- Message-Id: {p06020406c0e43c1982c3-at-[10.255.254.48]} } 8, 12 -- Date: Wed, 19 Jul 2006 15:44:43 -0400 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: shaenon-at-hotmail.com (by way of MicroscopyListserver) } 8, 12 -- Subject: viaWWW: LKB 8800A-NM Ultrotome III Problem } 8, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers==============================
-- ############################################################## 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 Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 12, 18 -- From bozzola-at-siu.edu Wed Jul 19 15:32:20 2006 12, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JKWKwA016808 12, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:32:20 -0500 12, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 12, 18 -- by abbmta2.siu.edu (Switch-3.1.9/Switch-3.1.7) with ESMTP id k6JKWGsJ019609 12, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:32:18 -0500 (CDT) 12, 18 -- Mime-Version: 1.0 12, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 12, 18 -- Message-Id: {p0611040bc0e4433498fd-at-[131.230.177.142]} 12, 18 -- In-Reply-To: {200607191945.k6JJjSfq022550-at-ns.microscopy.com} 12, 18 -- References: {200607191945.k6JJjSfq022550-at-ns.microscopy.com} 12, 18 -- Date: Wed, 19 Jul 2006 15:32:15 -0500 12, 18 -- To: Microscopy-at-msa.microscopy.com 12, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 12, 18 -- Subject: Re: [Microscopy] viaWWW: LKB 8800A-NM Ultrotome III Problem 12, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 12, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
We have an LKB Knifemaker Model 7801A that just stopped making knives. I had recently adjusted it, replaced the cutting wheel, lubricated a couple parts, etc., and it was making knives like gangbusters. About 3 weeks later the locking arm (the lever on the left that locks the scoring apparatus down) seemed to become looser and easily depresses to a lower point than it ever did before. It doesn't seem to be applying enough pressure on the cutting arm. It's still scoring, but turning the breaking wheel (the one on lower right that applies pressure to the scored glass) doesn't cause the glass to break.
Does anyone have a clue about how to fix this, or even what went wrong, from my probably garbled description? Are there any schematics out there that someone might be willing to share? Is there any relief from this heat?
Again, the only thing that seems to have changed is the locking arm feeling looser and it's not obvious to me how to adjust this.
Thanks for any advice you might have! I didn't think you could damage one of these things with an Abrams tank....
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well When Our Knifebreaker Works! 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
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Wed Jul 19 16:01:36 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 k6JL1ZCP027508 10, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 16:01:35 -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 -- Wed, 19 Jul 2006 16:01:30 -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: LKB Knifemaker on the skids 10, 23 -- Date: Wed, 19 Jul 2006 16:01:31 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E338E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: LKB Knifemaker on the skids 10, 23 -- thread-index: AcardoOcvNcWH+D4QRmtpEpa94Gopg== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:30.0992 (UTC) FILETIME=[82764F00:01C6AB76] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JL1ZCP027508 ==============================End of - Headers==============================
Many years ago, we developed a technique for etching PEEK for observation under TEM. But it would also work under a good modern SEM. Here are the two papers. If you want any more details, please contact me at R.H.Olley-at-reading.ac.uk .
On Crystallization Phenomena in PEEK Bassett,D.C., Olley,R.H., Al Raheil,I.A.M. Polymer, 1988, vol.29, pp.1745-1754
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
==============================Original Headers============================== 6, 23 -- From hinmeigeng-at-hotmail.com Wed Jul 19 16:01:45 2006 6, 23 -- Received: from bay0-omc2-s33.bay0.hotmail.com (bay0-omc2-s33.bay0.hotmail.com [65.54.246.169]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JL1j4E027635 6, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 19 Jul 2006 16:01:45 -0500 6, 23 -- Received: from hotmail.com ([64.4.56.23]) by bay0-omc2-s33.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Wed, 19 Jul 2006 14:01:44 -0700 6, 23 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 6, 23 -- Wed, 19 Jul 2006 14:01:43 -0700 6, 23 -- Message-ID: {BAY101-F1328BD6B77428118F7CCFFCA600-at-phx.gbl} 6, 23 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 6, 23 -- Wed, 19 Jul 2006 21:01:41 GMT 6, 23 -- X-Originating-IP: [86.128.172.175] 6, 23 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 6, 23 -- X-Sender: hinmeigeng-at-hotmail.com 6, 23 -- Reply-To: R.H.Olley-at-reading.ac.uk 6, 23 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 6, 23 -- To: Microscopy-at-MSA.Microscopy.Com 6, 23 -- Cc: R.H.Olley-at-reading.ac.uk 6, 23 -- Subject: Re: AskAMicroscopist: Observation of polymers morphology 6, 23 -- Date: Wed, 19 Jul 2006 21:01:41 +0000 6, 23 -- Mime-Version: 1.0 6, 23 -- Content-Type: text/plain; format=flowed 6, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:43.0828 (UTC) FILETIME=[8A1CED40:01C6AB76] ==============================End of - Headers==============================
What is happening is that the locking head is not locking it tight enough. This is caused by people (not you, of course, as I know that you know better) consistently overtightening the locking lever. If the lever is pushed all the way down so that the black ball is touching the base plate, it can no longer exert enough pressure to clamp the head in place. I believe that you can insert a metal washer on (the end of?) the shaft of the tightening rod and this will allow more pressure to be applied. But the big NO, NO here is (after it is fixed) not to push the arm all the way down to lock it in place. You stop pushing down when you feel resistance. Ideally, it should be safely locked with the black ball about 1 inch above the base plate.
You going to M&M?
JB
} We have an LKB Knifemaker Model 7801A that just stopped making knives. } I had recently adjusted it, replaced the cutting wheel, lubricated a } couple parts, etc., and it was making knives like gangbusters. About 3 } weeks later the locking arm (the lever on the left that locks the } scoring apparatus down) seemed to become looser and easily depresses to } a lower point than it ever did before. It doesn't seem to be applying } enough pressure on the cutting arm. It's still scoring, but turning the } breaking wheel (the one on lower right that applies pressure to the } scored glass) doesn't cause the glass to break. } } Does anyone have a clue about how to fix this, or even what went wrong, } from my probably garbled description? Are there any schematics out } there that someone might be willing to share? Is there any relief from } this heat? } } Again, the only thing that seems to have changed is the locking arm } feeling looser and it's not obvious to me how to adjust this. } } Thanks for any advice you might have! I didn't think you could damage } one of these things with an Abrams tank.... } } Cheers, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well When Our } Knifebreaker Works! } 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 } } } } } } } } ==============================Original Headers============================== } 10, 23 -- From TindallR-at-missouri.edu Wed Jul 19 16:01:36 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 k6JL1ZCP027508 } 10, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 } 16:01:35 -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 -- Wed, 19 Jul 2006 16:01:30 -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: LKB Knifemaker on the skids } 10, 23 -- Date: Wed, 19 Jul 2006 16:01:31 -0500 } 10, 23 -- Message-ID: } {91108EF9255B394CBF8B7E3789814A411E338E-at-UM-XMAIL08.um.umsystem.edu} } 10, 23 -- X-MS-Has-Attach: } 10, 23 -- X-MS-TNEF-Correlator: } 10, 23 -- Thread-Topic: LKB Knifemaker on the skids } 10, 23 -- thread-index: AcardoOcvNcWH+D4QRmtpEpa94Gopg== } 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 10, 23 -- To: {microscopy-at-microscopy.com} } 10, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:30.0992 (UTC) } FILETIME=[82764F00:01C6AB76] } 10, 23 -- Content-Transfer-Encoding: 8bit } 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k6JL1ZCP027508 } ==============================End of - Headers==============================
-- ############################################################## 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 Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 8, 18 -- From bozzola-at-siu.edu Wed Jul 19 16:20:17 2006 8, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JLKHj6015591 8, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 16:20:17 -0500 8, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 8, 18 -- by abbmta2.siu.edu (Switch-3.1.9/Switch-3.1.7) with ESMTP id k6JLKGIF001208 8, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 16:20:16 -0500 (CDT) 8, 18 -- Mime-Version: 1.0 8, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 8, 18 -- Message-Id: {p0611040ec0e450cdc906-at-[131.230.177.142]} 8, 18 -- In-Reply-To: {200607192103.k6JL3JkD031100-at-ns.microscopy.com} 8, 18 -- References: {200607192103.k6JL3JkD031100-at-ns.microscopy.com} 8, 18 -- Date: Wed, 19 Jul 2006 16:20:15 -0500 8, 18 -- To: Microscopy-at-msa.microscopy.com 8, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 8, 18 -- Subject: Re: [Microscopy] LKB Knifemaker on the skids 8, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 8, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
One last comment. Haskris offers, in addition to air- and water-cooled chillers, a heat exchanger for lower volume cooling needs. It depends solely on heat exchange between a supply of cool water and the closed system cooling the microscope. One significant limitation is that it depends on a cool water supply. Here in Texas, our supply of surface water in the summer can easily approach nearly 90F; too high a temperature for this heat exchanger to do much cooling. I suggest that you check out this heat exchanger if you work in an area where the tap water is cool to cold and the heat from your equipment is modest.
Regarding the need for an algaecide in chillers: We use opaque black hose for the cooling lines of our microscopes. This eliminates any source of light that can support growth of algae. Never use tubing of any kind. The hose is also heavier than tubing and resists hydrodynamic failure.
Practicing this over the past 20 years, we have never encountered problems with algae. Fill the chiller with deionized water and you're good to go.
Regards,
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
gary-at-gaugler.c om To gary.m.brown-at-exxonmobil.com 07/19/06 03:19 cc PM Subject [Microscopy] Re: Haskris chiller for Please respond Zeiss/LEO to gary-at-gaugler.c om
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Thanks for the on- and off-list replies. Very helpful as usual.
An additive is definitely needed. Haskris says no--just use distilled water. that does not work for long. I use 10% mix of Ethylene Glycol (1/2G to 4.5G distilled water). That works fine. I change the nylon and toilet paper filters about once a year and change out the water mix. It is also good practice to periodically check the nylon filter for signs of algae.
The R050 chillers for Zeiss/LEO are built with quite a few options. One of these is option G. This is a hot gas bypass system. The net effect of this is that the compressor runs continuously at the factory set temperature (65F). The temperature can be changed but the compressor will still run continuously. When Zeiss installed the system, they essentially bypassed this option. They changed the thermostat set point to 72F. Doing so causes the compressor to cycle. However, by using the turbine pump which is always running, compressor on/off is not seen at the SEM. The rationale for doing this change was to bring the chilled water closer to ambient temperature of the SEM to avoid condensation.
Zeiss uses a cooling design for its electronics that IMO is much more sophisticated than others. The consequence of this is that the chiller needs to have a larger reservoir (5G) and higher flow rate. but with pumps and chiller in separate rooms from the SEM, the heat output from the SEM is very low.
All feedback about Haskris has been hugely positive. My experiences with Haskris people is also very positive. So I will stick with them.
My problem turns out to be a blown compressor rather than a seized pump. The pump motor will not operate since the high temp sensor disables the motor. When the water cools down a little from the limit, the motor runs OK. The compressor is the problem. The windings are open and the current relay is basically vaporized. So the whole compressor has to be replaced. Has anyone seen this problem before? The only known rationale for this is low line voltage. In my case, the chiller is on a Liebert Nfinity 8KVA UPS. Since I always run in inverter mode, the chiller will always get perfect power. The UPS will take out any line sags or spikes.
The concept of a single point of failure is in operation....sigh.
gary g.
At 08:30 AM 7/19/2006, you wrote:
} Gary, I have a LEO/Zeiss SEM and also the same chiller. My previous } experience } with Haskriss chillers is that some sort of water additive must be } used to eliminate } bacteria growth and corrosion. When our 1550VP was first installed } we had problems } with clogging filters and thermal alarms on the SEM until we started } treating the water. } I have been using the same additive that our building chilled water } system engineers } use at about 4 ounces per tank. I have absolutely no idea what is in } it, but we no longer } have filter issues and the Haskriss runs continuously without any } sign of corrosion } anywhere. } } } } } } Does anyone know of an alternative to the Haskris R050 air cooled } chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at } least every year. The pump/motor unit will seize. Then the } compressor control circuitry will fry. } } Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I } don't see any other maker that can match this. } } If they are there, please advise. } } gary g.
Randy, We have a model 7800 knife maker. If these models are similar, here's what I think might have happened. When the unit was lubricated, it has allowed the adjusting collar on the tightening handle shaft to move (round collar with circular holes in it for a spanner wrench). Simply turning this adjusting collar to the correct position will correct the problem. Maybe removing some lubricant too!
Good luck, Kevin
Kevin Battjes Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Wednesday, July 19, 2006 5:07 PM To: battjes-at-impactanalytical.com
We have an LKB Knifemaker Model 7801A that just stopped making knives. I had recently adjusted it, replaced the cutting wheel, lubricated a couple parts, etc., and it was making knives like gangbusters. About 3 weeks later the locking arm (the lever on the left that locks the scoring apparatus down) seemed to become looser and easily depresses to a lower point than it ever did before. It doesn't seem to be applying enough pressure on the cutting arm. It's still scoring, but turning the breaking wheel (the one on lower right that applies pressure to the scored glass) doesn't cause the glass to break.
Does anyone have a clue about how to fix this, or even what went wrong, from my probably garbled description? Are there any schematics out there that someone might be willing to share? Is there any relief from this heat?
Again, the only thing that seems to have changed is the locking arm feeling looser and it's not obvious to me how to adjust this.
Thanks for any advice you might have! I didn't think you could damage one of these things with an Abrams tank....
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well When Our Knifebreaker Works! 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
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Wed Jul 19 16:01:36 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 k6JL1ZCP027508 10, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 16:01:35 -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 -- Wed, 19 Jul 2006 16:01:30 -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: LKB Knifemaker on the skids 10, 23 -- Date: Wed, 19 Jul 2006 16:01:31 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E338E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: LKB Knifemaker on the skids 10, 23 -- thread-index: AcardoOcvNcWH+D4QRmtpEpa94Gopg== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:30.0992 (UTC) FILETIME=[82764F00:01C6AB76] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JL1ZCP027508 ==============================End of - Headers==============================
==============================Original Headers============================== 22, 16 -- From battjes-at-impactanalytical.com Wed Jul 19 16:45:35 2006 22, 16 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 22, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JLjYkp003923 22, 16 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 16:45:34 -0500 22, 16 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 22, 16 -- id {M0R0NM63} ; Wed, 19 Jul 2006 17:45:33 -0400 22, 16 -- Message-ID: {089C7709BE9235448E3622C6F38D828F03EF6010-at-mitconexch.mitcon.org} 22, 16 -- From: "Battjes, Kevin" {battjes-at-impactanalytical.com} 22, 16 -- To: "'TindallR-at-missouri.edu'" {TindallR-at-missouri.edu} , 22, 16 -- "Microscopy List (E-mail)" {microscopy-at-microscopy.com} 22, 16 -- Subject: RE: [Microscopy] LKB Knifemaker on the skids 22, 16 -- Date: Wed, 19 Jul 2006 17:45:29 -0400 22, 16 -- MIME-Version: 1.0 22, 16 -- X-Mailer: Internet Mail Service (5.5.2656.59) 22, 16 -- Content-Type: text/plain; 22, 16 -- charset="iso-8859-1" ==============================End of - Headers==============================
On Jul 18, 2006, at 6:14 PM, gary-at-gaugler.com wrote:
} Does anyone know of an alternative to the Haskris R050 air cooled } chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at } least every year. The pump/motor unit will seize. Then the } compressor control circuitry will fry. } } Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I } don't see any other maker that can match this. } Dear Gary, Have you looked into the water-cooled Haskris units? Our two have had no problems since installation ~3 years ago, and the (larger) Haskris units on the HVEM at Albany NY gave virtually trouble-free performance for more than 20 years. We do and did regular preventive maintenance--checking flow rates, changing filters, adjusting pH anti-algae and anti-corrosion additives, etc. Here at Caltech, we have a house chilled water supply for cooling. Do you have enough air flow to cool your units properly, or do the specs indicate that you should use a larger unit? Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Jul 19 17:13:33 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 k6JMDWPa014798 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 17:13:33 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 7065C109B07 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:13:32 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id C16F6355A6 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:13:26 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200607190114.k6J1EXpI029214-at-ns.microscopy.com} 4, 22 -- References: {200607190114.k6J1EXpI029214-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {56d29ad5847931b087bff96111bbacad-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] 4, 22 -- Date: Wed, 19 Jul 2006 15:14:55 -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 earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Be sure to get a new Freon filter for your installation. Tecumseh makes the compressor and evaporator units. Current model is AKA4460YXD and should come with the current relay and start capacitor.
gary g.
At 01:28 PM 7/19/2006, you wrote: } Gary, } } We're in the process of replacing compressor and condenser unit because } this second compressor has worn out. We are changing to modern } refrigerant at the same time, hence the new condenser unit. Haskris } will tell you what model to get from local suppliers. You don't have to } buy it from them. The whole unit costs about $700. } } Ron L } } -----Original Message----- } From: gary-at-gaugler.com [mailto:gary-at-gaugler.com] } Sent: Wednesday, July 19, 2006 4:21 PM } To: lherault-at-bu.edu } Subject: [Microscopy] Re: Haskris chiller for Zeiss/LEO } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America
We've been looking at fungal hyphae in plant root tissues, and they contain unidentified compounds in what look like lipidic vesicles (ranging in size from tiny to a few microns in diameter) that are fluorescent under UV. After staining tissue plus fungus with toluidine blue, the compounds are substantially more fluorescent - blue-white fluorescence, which goes against any ancestral knowledge I've heard about - I usually use tol. blue to quench unwanted autofluorescence. Not only that, but the vesicle fluorescence increases substantially with increasing exposure to UV.
Any ideas as to what type of compound(s) these might be? Something with ring structures to absorb light - does animal lipofuchsin respond this way to UV?
thanks for any suggestions, cheers, Rosemray
Dr. Rosemary White rosemary.white-at-csiro.au CSIRO Plant Industry ph. 61 (0)2-6246 5475 GPO Box 1600 mob. 61 (0)402 835 973 Canberra, ACT 2601 fax. 61 (0)2-6246 5334 Australia
we have the exact same knife maker and the schematics + service instructions. There are only about six pages so I can easily scan them as JPEGs and email them off-list if you wish.
My only concern is that you should never lubricate the flat surfaces between the cutting head and the main stand with the handle. These rely on metal/metal contact.
If the head is no longer tightening at the right height there is a simple adjustment which involves removing a domed chrome screw at the front of the cutter stand near the clamping handle. Beneath this is a 3mm hexagonal screw which can be loosened slightly. When this is done you should be able to rotate a collar at the base of the clamping handle. This adjusts the position at which the locking lever clamps - so just try turning it a bit and test locking arm until it clamps at about the horizontal position. Then re-tighten the 3mm hexagonal screw and put the covering chrome screw back.
I can also send the schematics if you haven't already got them - LET ME KNOW, THOUGH. These details are under IV Troubleshooting Fault 6.
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: TindallR-at-missouri.edu
Dear group: - any chance someone out there has a copy of the operating manual for this model of centrifuge - it is refrigerated. Thanks Barbara
==============================Original Headers============================== 1, 18 -- From maloneyb-at-fiu.edu Thu Jul 20 10:18:27 2006 1, 18 -- Received: from smtp8.fiu.edu (smtp8.fiu.edu [131.94.79.25]) 1, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KFIQiU008830 1, 18 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Jul 2006 10:18:26 -0500 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu [131.94.95.156]) 1, 18 -- by smtp8.fiu.edu (MOS 3.7.1-GA) 1, 18 -- with ESMTP id CAI24841; 1, 18 -- Thu, 20 Jul 2006 11:18:23 -0400 (EDT) 1, 18 -- Message-ID: {44BF9F01.4020605-at-fiu.edu} 1, 18 -- Date: Thu, 20 Jul 2006 11:19:29 -0400 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 18 -- X-Accept-Language: en-us, en 1, 18 -- MIME-Version: 1.0 1, 18 -- To: microscopy-at-msa.microscopy.com 1, 18 -- Subject: Jouan CR4.11 centrifuge 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hello, I think the internal safety pressure seal on our critical point drier has burst. Does anyone know where I can get a PELCO CPD2 Critical point drier repaired at?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 2, 24 -- From gvrdolja-at-nature.berkeley.edu Thu Jul 20 11:29:44 2006 2, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 2, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KGTiaX020133 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 11:29:44 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 71648C1E8C 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 09:29:44 -0700 (PDT) 2, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 2, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 2, 24 -- with ESMTP id 17439-05 for {microscopy-at-microscopy.com} ; 2, 24 -- Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id 496F8C1E9B; Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 40A37C1E8C 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- Date: Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: microscopy-at-microscopy.com 2, 24 -- Subject: repair of critical point drier 2, 24 -- Message-ID: {Pine.SOC.4.64.0607200927360.11284-at-nature.Berkeley.EDU} 2, 24 -- MIME-Version: 1.0 2, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 2, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
I don't know how well etching will work with this material. Somebody else on the listserver will probably know better than me.
Alternative, you can stain with ruthenium tetroxide and examine using SEM, FE-SEM or TEM. Procedure is well described in following reference. This should be no problem if the components can be stained with RuO4.
GM Brown and JH Butler, New method for the characterization of domain morphology of polymer blends using ruthenium tetroxide staining and low voltage scanning electron microscopy (LVSEM), Polymer 38 (15), 3937-3945, 1997.
Regards,
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
"Sadhukhan, Pat" {SadhukhanPat-at- To BFUSA.com} {gary.m.brown-at-exxonmobil.com} cc
Dear Gary: Can we etch rubber particles from a cross-linked rubber/plastic composite ? If so, can you kindly suggest a procedure ? Regards, Pat Sadhukhan
Igauthier,
I expect that you will be disappointed. Typically, elucidation of the morphology of crystalline morphology of polymers generally requires etching or staining.
Regards,
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============================== 26, 19 -- From gary.m.brown-at-exxonmobil.com Thu Jul 20 12:21:03 2006 26, 19 -- Received: from hoespc02.exxonmobil.com (hoespc02.exxonmobil.com [192.67.48.39]) 26, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KHL3q6031222 26, 19 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 12:21:03 -0500 26, 19 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 26, 19 -- by hoespc02.exxonmobil.com (Switch-3.1.7/Switch-3.1.7) with ESMTP id k6KHL0Ob011027 26, 19 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 12:21:01 -0500 (CDT) 26, 19 -- In-Reply-To: {1178C664B84FD311AE9B0008C7A4F0BF05627D4E-at-serverex.bfs.com} 26, 19 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: Observation of Polymers morphology 26, 19 -- Importance: 26, 19 -- To: "Sadhukhan, Pat" {SadhukhanPat-at-BFUSA.com} , microscopy-at-microscopy.com 26, 19 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 26, 19 -- Message-ID: {OFEE8F83CD.0BCAA994-ON862571B1.005EBB62-862571B1.005F4D3C-at-exxonmobil.com} 26, 19 -- From: gary.m.brown-at-exxonmobil.com 26, 19 -- Date: Thu, 20 Jul 2006 12:20:58 -0500 26, 19 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652FP1HF193|March 26, 19 -- 02, 2006) at 07/20/2006 12:21:01 PM 26, 19 -- MIME-Version: 1.0 26, 19 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, July 20, 2006 at 13:24:06 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, U.S.A.
Title: Tissue Processing for EM
Question: We are considering purchasing a Lynx el tissue processor from Electron Microscopy Sciences. I would appreciate some opinions, pro and con concerning the Lynx. Also, we are looking at obtaining a vacuum oven from EMS, either the hydraulic thermostat controlled or the programmable microprocessor temperature controlled. Thank you all for your help.
We have routinely used the Lynx el Processor for approximately 15 years now. Extremely reliable. We have an established 14-vial processing protocol following fixation and rinsing of minced tissues: rinse, osmication, dehydration, PO, infiltration through pure resin - -~15ml each. Max time is 26 hours, though it's set to run overnight so whether you start at 8AM or 4PM, it's ready the next morning.
We routinely use the 4-well baskets. To maximize infiltration, don't overload wells. As we only embed 5 samples per tissue, I only include 5-6 minced pieces. On rare occasions, a sample will work it's way through the slots, but they never have crossed into other vials.
Cleaning is with lots of acetone. Osmium will react with metallic stem pieces and thus we rinse in acetone, then light wipe/scrub with Soft Scrub, and final sonication in diluted detergent.
The system is designed to process 7baskets x 4 wells each, but I typically will limit myself to 5 baskets (20 unique tissue samples) at a time. 3-well and 8-well baskets are also available.
The only other similar unit is from Leica at http://www.leica-microsystems.com/EM_Specimen_Prep
Best regards. Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
-----Original Message----- X-from: dfine-at-seton.org [mailto:dfine-at-seton.org] Sent: Thursday, July 20, 2006 2:58 PM To: Bobrowski, Walter
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, July 20, 2006 at 13:24:06 Remember to consider the Grade/Age of the student when considering the Question ------------------------------------------------------------------------ --- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ------------------------------------------------------------------------ ---
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, U.S.A.
Title: Tissue Processing for EM
Question: We are considering purchasing a Lynx el tissue processor from Electron Microscopy Sciences. I would appreciate some opinions, pro and con concerning the Lynx. Also, we are looking at obtaining a vacuum oven from EMS, either the hydraulic thermostat controlled or the programmable microprocessor temperature controlled. Thank you all for your help.
==============================Original Headers============================== 8, 12 -- From zaluzec-at-microscopy.com Thu Jul 20 13:51:20 2006 8, 12 -- Received: from [10.255.254.48] (msdvpn26.msd.anl.gov [130.202.238.90]) 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KIpIb9010602 8, 12 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 13:51:19 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: (Unverified) 8, 12 -- Message-Id: {p06020407c0e58118a310-at-[10.255.254.48]} 8, 12 -- Date: Thu, 20 Jul 2006 14:51:22 -0400 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- From: dfine-at-seton.org (by way of Ask-A-Microscopist) 8, 12 -- Subject: AskAMicroscopist: Tissue Processing for EM 8, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers============================== ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
==============================Original Headers============================== 26, 29 -- From Walter.Bobrowski-at-pfizer.com Thu Jul 20 14:42:56 2006 26, 29 -- Received: from gromsgoa01.pfizer.com (gromsgo.pfizer.com [148.168.34.153]) 26, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KJguIF021850 26, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 14:42:56 -0500 26, 29 -- Received: from groamrexc01.amer.pfizer.com (groamrexc01.amer.pfizer.com [172.30.8.168]) 26, 29 -- by gromsgoa01.pfizer.com (8.13.4/8.13.4) with ESMTP id k6KJdwA6006752; 26, 29 -- Thu, 20 Jul 2006 15:39:58 -0400 26, 29 -- Received: from groamrexc02.amer.pfizer.com ([172.30.8.169]) by groamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 26, 29 -- Thu, 20 Jul 2006 15:42:53 -0400 26, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 26, 29 -- Thu, 20 Jul 2006 15:42:53 -0400 26, 29 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 26, 29 -- Content-class: urn:content-classes:message 26, 29 -- MIME-Version: 1.0 26, 29 -- Content-Type: text/plain; 26, 29 -- charset="us-ascii" 26, 29 -- Subject: RE: [Microscopy] AskAMicroscopist: Tissue Processing for EM 26, 29 -- Date: Thu, 20 Jul 2006 15:42:31 -0400 26, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D07407EBB-at-anaamrexm01.amer.pfizer.com} 26, 29 -- X-MS-Has-Attach: 26, 29 -- X-MS-TNEF-Correlator: 26, 29 -- Thread-Topic: [Microscopy] AskAMicroscopist: Tissue Processing for EM 26, 29 -- Thread-Index: AcasLmH+Jm0RptsvTWmGLL8i9w1EMgABEAxg 26, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 26, 29 -- To: {dfine-at-seton.org} , {Microscopy-at-microscopy.com} 26, 29 -- X-OriginalArrivalTime: 20 Jul 2006 19:42:53.0606 (UTC) FILETIME=[B1181460:01C6AC34] 26, 29 -- X-Proofpoint-Spam-Reason: safe 26, 29 -- Content-Transfer-Encoding: 8bit 26, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6KJguIF021850 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, July 20, 2006 at 15:05:16 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, USA
Title: Leica EMPT
Question: I would appreciate some information from the EM labs using the Leica EMTP. Is it user-friendly and reliable? Would you purchase it again? Were there any problems or issues with it? Do you run 2 different processing programs together at the same time? Do you run separate procedures for kidney and muscle biopsies? Are different procedures run for each different tissue? Thanks for the replies.
I was wondering what that slotted screw was for and even had it out, but didn't notice the hex screw underneath. The arm is now adjusted to lock just above horizontal.
It's still a little off, because I have to apply way too much pressure on the locking lever to keep it from riding up when the breaking wheel is turned. And that's a Bad Thing. But we are making knives and I'll keep playing with the fine-tuning.
Thanks for all the very informative responses.
Randy
Randy Tindall 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
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jul 20 16:53:44 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KLrh6j012049 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 16:53:43 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 20 Jul 2006 16:53:43 -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: LKB Knifemaker 10, 23 -- Date: Thu, 20 Jul 2006 16:53:43 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E3394-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: LKB Knifemaker 10, 23 -- thread-index: AcasRve4gW1T67jgRWG66JuqMTWodg== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 20 Jul 2006 21:53:43.0476 (UTC) FILETIME=[F7FB2F40:01C6AC46] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6KLrh6j012049 ==============================End of - Headers==============================
That software would be BTVPro from http://www.bensoftware.com/ timelapse, motion detection, frame averaging, file export and many other features On Mar 1, 2005, at 8:04 AM, James Pawley wrote:
} } } ---------------------------------------------------------------------- } -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } } --------------------------------------------------------------------- } } --------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } } ---------- } } } } Roger; } } } } You might find some past discussions, or care to raise } } this issue again in the Yahoo Microscope group. Both } } issues (LED Illumination and inexpensive camera } } options) have been discussed there in the past. That } } group is more focused (no pun intended) on LM. It is } } also more of an amateur orientated group, although } } many of the prime contributors are professionals (or } } retirees). As a result of the audiance, a number of } } low cost but "workable" alternatives are discussed. } } } } The latest option for cameras seems to be using "Web } } Cams", either out of the box (with standard occulars), } } or by removing the lens assemblies and using a Photo } } or Projection eyepiece. } } } } I've played with LEDs myself, and have experienced } } results similar to what you describe (OK, but not } } spectacular). For me, it is more a matter of a light } } source that can be used "in the field". } } } } John Raffensperger, Jr. } } Beaver Dam, Wisconsin } } } } Hi all, } } I hope that it is not too commercial to tell you that, in the next } edition of The Handbook, there will be a fairly complete discussion } of how LEDs might be used in microscopy in the chapter on Non-laser } Light Sources by Andreas Nolte from Zeiss. At least it shows that } the idea has promise. } } And on the cheap CCD front, I have had fair success hooking the } iSight camera from apple (~$140, 640x480, Firewire) to a Zeiss } scope by the simple expedient of removing the rubber "eye-glasses" } protector gasket on the normal 10x high-eyepoint occular and } holding the camera right in front of it with a plastic collar. It } covers a little more than the whole field of view (some black } corners). Although I haven't used it, I am told that one can use } $30 "surveillance" software to record time-lapse sequences on a Mac } using this camera. Might be neat for non-microscope uses too. } } Cheers, } } Jim P. } -- } ********************************************** } Prof. James B. Pawley, Ph. } 608-263-3147 Room 223, Zoology Research Building, } FAX 608-265-5315 } 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU } 3D Microscopy of Living Cells Course, June 11-23, 2005, UBC, } Vancouver Canada } Info: http:// www.3dcourse.ubc.ca/ Applications due by March } 15, 2005 } } Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
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I have put my answers alonside your questions.
} Email: dfine-at-seton.org } Name: Diann Fine } } Organization: Brackenridge Hospital } } Education: Graduate College } } Location: Austin, Texas, USA } } Title: Leica EMPT } } Question: I would appreciate some information from the EM labs using the } Leica EMTP. Is it user-friendly and reliable?
} YES - on both counts, based on our experience of using it for a year for } acrylic and epoxy resin processing for high res LM and TEM respectively.
} Would you purchase it again?
} YES - in my view it is by far the best processor on the market for dual } purpose high throughput LM resin histology and TEM
} Were there any problems or issues with it?
} YES - the turntable height had to be adjusted to a lower position as the } vials were catching on the bottom of the cooling jacket. This has been the } only service callout in the year we have been using it.
} Do you run 2 different processing programs together at the same time?
} I don't think that it is possible to do this.
} Do you run separate procedures for kidney and muscle biopsies? YES
} Are different procedures run for each different tissue? NO
We have 2 basic schedules for both epoxy and acrylic resin processing. A relatively short (working day) schedule for small, soft easy to infiltrate tissues such as renal biopsies and longer (overnight) schedule for muscle and other tissues that are more difficult to infiltrate.
} Thanks for the replies.
Get back to me if you want a copy of our protocols.
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Email: nessonm-at-onid.orst.edu Name: Michael Nesson
Organization: Oregon State University EM Facility
Title-Subject: [Filtered] X-ray Detector Repair
Question: We have a blown window on our Kevex detector on our FEI CM12 TEM. Please respond off-line to me if you have had either exceptionally good or exceptionally bad experiences with the three companies that advertise in "Microscopy Today": namely Advanced Analysis Technologies, e2v Scientific Instruments (formerly Gresham), and MAX DETECTOR Repair Group LLC. Again, please respond to me directly (nessonm-at-onid.orst.edu), rather than to the Listserv, so as to reduce the possibility of flame wars. TIA, Mike Nesson
Michael H. Nesson, Ph.D. Electron Microscope Facility 1078 Cordley Hall Oregon State University Corvallis, OR 97331 (541) 737-5245
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Title-Subject: [Filtered] Re: [Microscopy] viaWWW: LKB 8800A-NM Ultrotome III Problem
Question: Following up on my recent LKB III Microtome troubles...
First, thank you to all that responded, your comments & suggestions were very helpful.
So, after investigating further this is what I have concluded: 1. The cord/string that raises the specimen arm is intact and functional. When I lift the arm up with my hand it moves as it should. Also, when I move the arm into "lock position", the arm is lifted up. 2. The belt in my control unit that is attached to the manual control knob is intact and moves as it should when I turn the knob. However, the specimen arm does not move. 3. There is no resistance when I move the manual control know, thus I don't think the potentiometer is seized up.
Also - when I switch to "auto" mode using the manual control knob, nothing happens. There is still a connection with the control unit however as the lamp still turns on.
Any further suggestions would be greatly appreciated!
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Email: javaidqazi-at-kemet.com Name: javaid qazi
Title-Subject: [Filtered] microscopy training
Question: Apart from Lehigh is there any other place which does training for SEM users.
The College of Microscopy located in Westmont IL offers SEM courses twice a year. Check out their website
www.collegeofmicroscopy.com
Regards
Alan
At 10:24 AM 7/21/2006 -0500, javaidqazi-at-kemet.com wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Yes, the McCrone group in Westmount, Il, outside of Chicago. They're nice people.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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javaidqazi-at-kemet. com To: frank.karl-at-degussa.com cc: 07/21/2006 11:24 Subject: [Microscopy] viaWWW: microscopy training AM Please respond to javaidqazi
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Email: javaidqazi-at-kemet.com Name: javaid qazi
Title-Subject: [Filtered] microscopy training
Question: Apart from Lehigh is there any other place which does training for SEM users.
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Email: hu.duan-at-averydennison.com Name: Hu
Title-Subject: [Filtered] question regarding roughness in large area
Question: Dear colleagues:
I have an application to quantitatively monitor the roughness/topology change over a relatively large area (3-5cm*3-5cm). The surface is not uniform. Thus, monitoring a large area is necessary. The conventional AFM, optical profilometry techniques suffer due to their limited scanned areas. Even with image stitching, the results are not satisfied due to complication of tilt, plane fit over such large area. I am wondering is there such technique/instrumentation to do such kind of work? Also I know mechanical profiler like Dektak can do similar work. However, our coating is relatively soft, usual stylus applies too much force and often creates scratches. Are there any optical methods for such application? Thank you for your attention and suggestions.
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Email: Sandow50-at-yahoo.com Name: Stanton Dowd
Organization: VCU
Title-Subject: [Filtered] Deton DV-502A High Vac issues
Question: I have been using an old DV-502A for routine carbon coating for the past four years without any significant problems, but recently I have had problems getting it down to operating vacuum (~1x10^-4) without using liquid nitrogen in the trap. Until about a month ago I've never needed to use the trap to achieve a much stronger vacuum. The problem seems to be getting worse without any overt reason. I've changed out the mechanical pump and damn near every seal in the thing in the past few weeks with no effect. As far as I can tell there is no significant leak anywhere, it simply seems that the diffusion pump efficiency has tanked. Using nitrogen in the trap I can still pump down to well under 1x10^-5 Torr, but without it the diffusion pump barely works at all. Assuming that there are no leaks, that the mechanical pump is working as it should, that the diffusion oil is clean and at the appropriate level and that the tree is intact; what can effect diffusion pumping efficiency? What I am left to wonder about is the column temperature gradient. I have the model with the 3.5" column if that matters. The peak temp is about one inch from the bas a reads at around 365C. The temp drops quickly over the first three inches and reads around 85C at the first cooling coil. There is a smooth gradient up the column to the top where the temp is right at 30C. I never bothered to take readings when it was working so I don't know if these numbers are normal or not. We are using tap water as the coolant and draining it into the sink. With the weather as warm as it has been I wonder if the influent temp is too warm to cool it adequately. For what its worth, the influent is 25.5C with a flow rate of 1.65 L/min and an effluent temp of 28C.
Any help that you guys can give would be greatly appreciated.
Water T is good.You don't want it colder than 15C and hotter than 30C (at supply end), optimum at return end is between 30 and 35C. Actual numbers may depend on flow rate, but it is good to stay within above limits. Speaking of leaks- TC (thermocouple) sensor reads below 20 mTorr with main valve open, there is no leaks. High vacuum leak always results in elevated backing pressure of the diff. pump. No elevated back pressure means no leak. I assume that mech. pump works well, which is tested by pumping lines only with all valves closed.
The potential diff. pump problems are:
1) Denton diff. pumps have water cooling tube coil cemented to the pump with heat conductive cement. More classy and expensive pumps such as Edwards or Varian have cooling tubing welded to pump column. Cement eventually cracks, and stops transferring heat. You need to examine it very closely and try to move the water cooling coil to see if it is still bonded to the pump. If it isn't, then either re-cement, or take pump off the unit, disassemble, clean it, and braze or solder water tubing to the pump column.
2) Depleted or lost diff. pump fluid. Check and replace or re-fill as necessary.
3) Silicone diff. pump fluid (such as Dow Corning 702 or 704 or 705) that is typically used by Denton may partially polymerize inside the pump. It will look clear and otherwise normal, with one difference. Touch the cooling tower inside the pump, and you will feel sticky layer - feels like sticky packaging tape or note stickers from Office Depot. Layer is clear and invisible, but easily detectable by touch. If this happened, pump is dead, even though fluid level is normal and all looks clean. Discard old fluid, disassemble the pump, and clean in very diligently. Acetone rub works for sticky stuff, but solid deposits (if present) must be removed with wire brush or sandblasting.
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: {Sandow50-at-yahoo.com} To: {vitalylazar-at-att.net} Sent: Monday, July 24, 2006 8:33 PM
Stanton,
Remove the DP and check that the DP oil is good. It might be 'cracked'. Replace it if you need to do that. I would do it anyway.
We had a very similar experience to yours on a JSM 35 SEM. The factory spent 3 weeks looking for a vacuum leak with a helium leak detector and examined every o-ring in the system. No good. A different serviceman came in and found the problem in one hour. The one terminal connection on a three month old DP heater was corroded. It was installed by the first serviceman to solve the same problem months ago. A new heater and terminal connection restored the pumping capacity to normal. Apparently the terminal limited the current to the DP heater, the DP got hot, but it never got hot enough to restore full vacuum pumping speed. LN2 helped but we knew the non-LN2 vacuum was bad. This seems to be a carbon copy of your problem and your assumptions of what is working properly.
You asked, "What can effect diffusion pumping efficiency?" The DP chimney port is not lined up with the rotary pump port. The DP oil is cracked. The DP oil is less than normal. The DP heater is defective in some way. Insufficient DP heating. A bad DP heater connection. Hot or limited cooling water. The "loss of cooling water" safety switch on the DP is opening at a lower temp. Cracked or leaking o-ring at the DP interface with the LN2 trap.
Paul
} Email: Sandow50-at-yahoo.com } Name: Stanton Dowd } } Organization: VCU } } Title-Subject: [Filtered] Deton DV-502A High Vac issues } } Question: I have been using an old DV-502A for routine carbon coating for the past four years without any significant problems, but recently I have had problems getting it down to operating vacuum (~1x10^-4) without using liquid nitrogen in the trap. Until about a month ago I've never needed to use the trap to achieve a much stronger vacuum. The problem seems to be getting worse without any overt reason. I've changed out the mechanical pump and damn near every seal in the thing in the past few weeks with no effect. As far as I can tell there is no significant leak anywhere, it simply seems that the diffusion pump efficiency has tanked. Using nitrogen in the trap I can still pump down to well under 1x10^-5 Torr, but without it the diffusion pump barely works at all. } Assuming that there are no leaks, that the mechanical pump is working as it should, that the diffusion oil is clean and at the appropriate level and that the tree is intact; what can effect diffusion pumping efficiency? What I am left to wonder about is the column temperature gradient. I have the model with the 3.5" column if that matters. The peak temp is about one inch from the bas a reads at around 365C. The temp drops quickly over the first three inches and reads around 85C at the first cooling coil. There is a smooth gradient up the column to the top where the temp is right at 30C. I never bothered to take readings when it was working so I don't know if these numbers are normal or not. We are using tap water as the coolant and draining it into the sink. With the weather as warm as it has been I wonder if the influent temp is too warm to cool it adequately. For what its worth, the influent is 25.5C with a flow rate of 1.65 L/min and an effluent temp of 28C. } } Any help that you guys can give would be greatly appreciated. } } Sincerely, } Stanton }
==============================Original Headers============================== 8, 26 -- From beaurega-at-westol.com Tue Jul 25 07:51:27 2006 8, 26 -- Received: from smtp-gateway-6.winbeam.com (smtp-gateway-6.winbeam.com [64.84.97.71]) 8, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PCpRx2022896 8, 26 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 07:51:27 -0500 8, 26 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 8, 26 -- by smtp-gateway-6.winbeam.com (8.13.1/8.12.8) with SMTP id k6PCpKTj013505 8, 26 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 08:51:21 -0400 8, 26 -- Received: (qmail 22938 invoked by uid 89); 25 Jul 2006 12:51:09 -0000 8, 26 -- Received: from pitts-69-72-13-84.dynamic-dialup.coretel.net (HELO millenium) (69.72.13.84) 8, 26 -- by mail.winbeam.com with SMTP; 25 Jul 2006 12:51:09 -0000 8, 26 -- Message-Id: {3.0.6.32.20060725085246.007d7560-at-pop3.norton.antivirus} 8, 26 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus (Unverified) 8, 26 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 8, 26 -- Date: Tue, 25 Jul 2006 08:52:46 -0400 8, 26 -- To: microscopy-at-microscopy.com 8, 26 -- From: Beaurega {beaurega-at-westol.com} 8, 26 -- Subject: Re: [Microscopy] Re: viaWWW: Deton DV-502A High Vac issues 8, 26 -- In-Reply-To: {200607250217.k6P2HALh003819-at-ns.microscopy.com} 8, 26 -- Mime-Version: 1.0 8, 26 -- Content-Type: text/plain; charset="us-ascii" 8, 26 -- X-Winbeam-MailScanner-Information: smtp-gateway-6.winbeam.com - Please contact Technical Support for more information 8, 26 -- X-Winbeam-MailScanner: Found to be clean [smtp-gateway-6.winbeam.com] (courtesy of Winbeam) 8, 26 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, 8, 26 -- score=0.276, required 6, autolearn=disabled, 8, 26 -- MAILTO_TO_SPAM_ADDR 0.28) 8, 26 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
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Email: khibbs-at-wistar.org Name: Kristine Hibbs
Organization: The Wistar Institute
Title-Subject: [Filtered] Announcement of Job Opening
Question: POSITION IN MICROSCOPY
THE WISTAR INSTITUTE
Research Assistant
The Wistar Institute is an independent nonprofit biomedical research institution dedicated to discovering the causes and cures for major diseases, including cancer, cardiovascular disease, autoimmune disorders, and infectious diseases, including AIDS and influenza. Founded in 1892 as the first institution of its kind in the nation, The Wistar Institute today is a National Cancer Institute-designated Cancer Center focused on basic and translational research. Discoveries at Wistar have led to the creation of vaccines for such diseases as rabies, rubella, and rotavirus; significant insights into the mechanisms of skin, brain, breast, lung, and prostate cancers; and the development of monoclonal antibodies and other significant research technologies and tools. Located on the campus of the University of Pennsylvania, The Wistar Institute maintains its status as an independent research center while enjoying a close working relationship with the University of Pennsylvania, ChildrenÇs Hospital of Philadelphia, and other medical research organizations in the greater Philadelphia area.
The Wistar Institute has an opening for a Research Assistant who will manage the multi (two)-photon microscope at the Institute. The individual will operate the microscope and interact with users. BachelorÇs degree required, but MasterÇs degree preferred. Background in microscopy required. Experience with confocal laser and/or two-photon microscopy required. Previous lab experience desired. Ability to problem solve and work independently required.
We offer an excellent benefits package, including tuition assistance. We do not offer relocation assistance. To apply, visit our webpage at http://www.wistar.org/humanresources/employment.htm and submit your resume and cover letter stating salary requirements and previous experience with confocal laser and/or two-photon microscopy.
The Wistar Institute, HR Dept., 3601 Spruce St., Philadelphia, PA 19104 EOE/AA/M/F/D/V.
For more information about The Wistar Institute visit our website at www.wistar.org.
Kristine Hibbs Human Resources Specialist The Wistar Institute (215) 898-3766 Khibbs-at-wistar.org
I am trying to get some paraformaldehyde into solution. I have probably done this several thousand times in my career without a problem. I am using a fairly new bottle of paraformaldehyde (prill type) and putting 2% in water (deionized or distilled - both have the same problem). I heat to 55 C (not over 60 C) with vigorous stirring, add a few drops of 1 N NaOH and the bulk goes into solution. The problem is there is a significant amount of flocculent material which doesn't go into solution. I made up a fresh batch of NaOH and there was no improvement. Has anyone experienced a similar problem?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
As suggested by others, Stanton's problem with his diffusion pump may very well be due to insufficient heat input from the DP's heater. If you look on the manufacturer's tag that is probably attached to the pump somewhere, you should find the value of the heater input in Watts. Contact your local electrician and he should have a gadget that he can simply clip around the wire leading into the heater and measure the current flowing through it. Current (Amps) x voltage (volts) = Watts. If the value measured doesn't equal the value on the pump's tag, you know you have a problem with the heater. Then, disconnect the heater wires and measure the resistance of the heating elements to see if one is burned out. Making a few measurements like this should give you an indication of whether or not there is a problem with the heater. If not, check the other factors mentioned. Good luck, WCB -- 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============================== 1, 14 -- From bigelow-at-engin.umich.edu Tue Jul 25 11:07:33 2006 1, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PG7Xd7025083 1, 14 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 11:07:33 -0500 1, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- by srvr22.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k6PG7WRg018471 1, 14 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 12:07:32 -0400 (EDT) 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06210200c0ebef5a2b80-at-[141.212.131.221]} 1, 14 -- Date: Tue, 25 Jul 2006 12:07:31 -0400 1, 14 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 1, 14 -- Subject: [Microscopy}RE: Diff. Pump Problem 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Hello everybody, I used JB-4 resin for embedding of fish larvae. Tissue was fixed in 2% glut, dehydrated in ethanol and infiltrated with ethanol/resin mixture (50-50, 75-25, 100). Last infiltration was overnight. The resin was polymerized at RT under a vacuum. Blocks seem to be hard but I can not cut them on glass knife (I cut 3 micron section). The resin either curls up and/or sticks to the surface of a knife. I stored JB-4 at +4C, but it was brought to a RT before infiltration and embedding. I noticed that one of the components (accelerator) expired May 06. Do you have any suggestions/advice how to improve the procedure so the blocks can be cut? Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Tue Jul 25 13:10:59 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PIAxVm005474 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 13:10:59 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1G5RMw-0001VW-02 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 15:10:58 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 25 Jul 06 15:10:58 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 25 Jul 06 15:10:43 -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: Tue, 25 Jul 2006 14:57:49 -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: JB-4 resin problems 1, 22 -- Message-ID: {44C6316C.19011.108EB5-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
} As suggested by others, Stanton's problem with his diffusion } pump may very well be due to insufficient heat input from the } DP's heater. ...
I remember a similar problem, which could have well been the heater output ... But in this case it was because I had changed the DP fluid to another that had a higher boiling pt. The heater did work ... But it sure was sluggish in the beginning.
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, NL
==============================Original Headers============================== 7, 21 -- From michael-at-shaffer.net Tue Jul 25 13:39:13 2006 7, 21 -- Received: from ws6-5.us4.outblaze.com (ws6-5.us4.outblaze.com [205.158.62.152]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6PIdCN0016239 7, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 13:39:12 -0500 7, 21 -- Received: (qmail 24459 invoked from network); 25 Jul 2006 18:39:13 -0000 7, 21 -- Received: from unknown (HELO rarewolf) (michael-at-shaffer.net-at-205.251.84.119) 7, 21 -- by ws6-5.us4.outblaze.com with SMTP; 25 Jul 2006 18:39:12 -0000 7, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 21 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 21 -- Cc: {bigelow-at-engin.umich.edu} 7, 21 -- Subject: RE: [Microscopy] [Microscopy}RE: Diff. Pump Problem 7, 21 -- Date: Tue, 25 Jul 2006 16:09:12 -0230 7, 21 -- Message-ID: {000b01c6b019$a08639e0$4701a8c0-at-rarewolf} 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 11 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 7, 21 -- Thread-Index: AcawBILzsLXAAdW2Qhi/oRSyFMrupwAFIf3g 7, 21 -- In-Reply-To: {200607251608.k6PG81A3025848-at-ns.microscopy.com} ==============================End of - Headers==============================
If you are heading to Chicago for the M&M meeting and you have an interest in High Pressure Freezing you are cordially invited to attend the following special events:
1. Sunday, July 30, 12:00 - 5:00pm, High Pressure Freezer User Group Meeting. Location: Sheraton Chicago Hotel & Towers, 301 East North Water St, Parlor E Buffet lunch served at 12:00 noon followed by a talk on High Pressure Freezing Techniques & Methods by Dr. Andres Kaech, Life Science Applications Manager, Bal-Tec AG at 1:00pm. Following this presentation the floor will be open for group discussion, questions & answers and/or individual presentations if anyone has a special topic to present to the group.
2. Thursday/Friday, August 3/4, small group High Pressure Freezing Workshop at the University of Chicago. This "hands-on" workshop hosted by the Dept of Molecular Genetics and Cell Biology and the Bioscience Division will be conducted in the laboratory of Dr. Joe Austin utilizing the Departments Bal-Tec HPM 010 High Pressure Freezer. Location: Lectures at 9:30am Thursday in CIS 400B (Center for Integrative Science, 4th floor) 929 E. 57th St (corner of 57th and Drexel) followed by lunch. Practical demonstrations and freezing in the lab, CIS ESB 06 (basement) in the afternoon and Friday morning starting at 9:00am The University of Chicago campus is approximately 25 minutes by car from the Navy Pier area. Please go to http://maps.uchicago.edu/westquad/irb.html to view a map of the building and nearby parking.
There is no charge to attend either of these meetings and lunch/refreshments are compliments of Bal-Tec RMC
See you in Chicago.
Dave Roberts Bal-Tec RMC Boeckeler Instruments Inc 4650 S. Butterfield Drive Tucson, AZ 85714 Tel: 520-745-0001 Fax: 520-745-0004 www.baltec-rmc.com
==============================Original Headers============================== 10, 20 -- From dave-at-boeckeler.com Tue Jul 25 16:12:04 2006 10, 20 -- Received: from txslsmtp2.vzwmail.net (txslsmtp2.vzwmail.net [66.174.85.156] (may be forged)) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PLC4fG029472 10, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 16:12:04 -0500 10, 20 -- Received: from tweedledee (smtp.vzwmail.net [66.174.85.25]) 10, 20 -- (authenticated bits=0) 10, 20 -- by txslsmtp2.vzwmail.net (8.12.9/8.12.9) with ESMTP id k6PLBpxF016056 10, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 21:12:01 GMT 10, 20 -- Message-Id: {200607252112.k6PLBpxF016056-at-txslsmtp2.vzwmail.net} 10, 20 -- From: "Dave Roberts" {dave-at-boeckeler.com} 10, 20 -- To: {microscopy-at-msa.microscopy.com} 10, 20 -- Subject: High Pressure Freezing Events in Chicago 10, 20 -- Date: Tue, 25 Jul 2006 14:11:48 -0700 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, Build 11.0.5510 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 10, 20 -- Thread-Index: AcawLvDyuWocdYaOST6Upc4KaWZBAA== ==============================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 martin.roe-at-nottingham.ac.uk as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: martin.roe-at-nottingham.ac.uk Name: Martin Roe
Organization: Nottingham University
Title-Subject: [Filtered] JEOL 6000-series keyboard for 6400 SEM
Question: Dear Listservers, Does anyone have an old and spare JEOL 6000-series keyboard for a JEOL 6400 SEM (plugs into a DIN socket below the filament control)? Willing to pay, if necessary. Thanks Martin Roe
Martin Roe EM and XPS Technical Support Wolfson Building School of Mechanical, Materials, Manufacturing Engineering Nottingham University University Park Nottingham NG7 2RD England, UK
Yes, I have had a similar problem and found that I had to increase the temperature to 60 C, having found exactly as you describe at even a slightly lower temperature.
Given the hassle, I now by it in in sealed ampoules.
Hope this works for you,
Alastair
At 10:52 25/07/2006 -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============================== 12, 18 -- From a.d.mckinnon-at-abdn.ac.uk Wed Jul 26 03:24:36 2006 12, 18 -- Received: from mailhub1.abdn.ac.uk (mailhub1.abdn.ac.uk [139.133.7.28]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6Q8OZwJ004959 12, 18 -- for {Microscopy-at-Microscopy.Com} ; Wed, 26 Jul 2006 03:24:35 -0500 12, 18 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 12, 18 -- by mailhub1.abdn.ac.uk with esmtp (Exim 4.52) 12, 18 -- id 1G5ehB-00018h-Tx; Wed, 26 Jul 2006 09:24:46 +0100 12, 18 -- Message-Id: {5.2.1.1.0.20060726091621.01151600-at-mailms.abdn.ac.uk} 12, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 12, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 12, 18 -- Date: Wed, 26 Jul 2006 09:24:31 +0100 12, 18 -- To: phillipst-at-missouri.edu 12, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 12, 18 -- Subject: Re: [Microscopy] paraformaldehyde problem 12, 18 -- Cc: Microscopy-at-Microscopy.Com 12, 18 -- In-Reply-To: {200607251552.k6PFqnwt023624-at-ns.microscopy.com} 12, 18 -- Mime-Version: 1.0 12, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
returned from an interesting conference(ULTRAPATH XIII, Rapid City, SD.,USA) and a short vacation, I have found your query concerning the making of paraformaldehyde stock solution.
I do not know about the source of your recipe for this, especially to heat the crude, hydrous solution with the PFA-powder ONLY unto "} 55 { degr.C (not above 60 degr.C)".
All the } old fashioned { sources about PFA-fixative I know (e.g. GEYER G. ed., Ultrahistochemie / Ultrahistochemistry G.FischerVerlag Stuttgart Germany 1973, or: PLATTNER H, Zingsheim HP. eds, Elektronenmikroskopische Methodik in der Zell- und Molekularbiologie, G.Fischer Verl. Stuttgart 1987, certainly as well as other [engl.] publications) state:
".....heat solution to approx. 65- } } 70 { { degr.C (but let NOT BOIL), - thoroughly mix up, add 1-3 drops [*whatever this means, depends perhaps on the dispensing device you use....*]
of 1 N NaOH......(and) until solution clears up....." [* I generally add drop after drop, waiting after each drop several minute wether the solution clears up or not*]
Perhaps the problem you face is a problem of incomplete dissolution due to a too low temperature.
IMO you would not need to discard such a solution, just filtering yields a fixative which perhaps has a slightly lower percentage in FA-concentration than you intended.
Another poblem would be precipitation of flocculent material after mixing the FA-solution with glutaraldehyde: this IMO points to a perhaps degraded or low quality batch of the concentrated GA-solution.
Greetings and best regards,
Wolfgang Muss SALZBURG, AUSTRIA
---------- Von: phillipst-at-missouri.edu[SMTP:phillipst-at-missouri.edu] Antwort an: phillipst-at-missouri.edu Gesendet: Dienstag, 25. Juli 2006 17:51 An: W.Muss-at-salk.at Betreff: [Microscopy] paraformaldehyde problem
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Biology Department at St. Francis Xavier University, seeks applications for a full-time Research Technician to interact with a group of ten research active, faculty and their students. The successful applicant should have a minimum of a B.Sc. preferably with experience operating laser confocal and electron microscopes. Experience in specimen preparation and image analysis are desired assets. The applicant must have a willingness to develop skills and instrumentation expertise in other areas, as required.
The Biology Department is housed in a newly-renovated building with microscope facilities that include a Philips EM410 TEM, a Jeol JSM 5300 SEM and an Olympus FV300 CLSM. The Biology Department has a general interest in aquatic biology, including invertebrate reproduction, vertebrate physiology, invasive algal and crustacean species, carbon fixation in cyanobacteria, marine algal symbiosis, as well as expertise in cell biology, microbiology and biomechanics.
This is a full time position with funding including benefits for 1 year initially followed by 2 additional years upon successful review of the applicant. Applications will be considered beginning August 15th, 2006 and the competition will remain open until the position is filled. Interested individuals should forward a CV outlining relevant experience, and the names and addresses of three references to:
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St. Francis Xavier University is committed to employment equity and welcomes applications from all qualified women and men, including aboriginal people, members of visible minorities, and persons with disabilities.
==============================Original Headers============================== 10, 20 -- From dmorriso-at-stfx.ca Wed Jul 26 07:37:27 2006 10, 20 -- Received: from xmail1.stfx.ca (xmail1.stfx.ca [141.109.221.24]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QCbJDk029410 10, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 07:37:24 -0500 10, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 20 -- Content-class: urn:content-classes:message 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="iso-8859-1" 10, 20 -- Subject: Job Posting: Research Technician in Microscopical Analysis 10, 20 -- Date: Wed, 26 Jul 2006 09:34:49 -0300 10, 20 -- Message-ID: {D1DA484CA5D7D5499AB6BBB4BCA4291750475F-at-demetrius.ad.stfx.ca} 10, 20 -- X-MS-Has-Attach: 10, 20 -- X-MS-TNEF-Correlator: 10, 20 -- Thread-Topic: Job Posting: Research Technician in Microscopical Analysis 10, 20 -- Thread-Index: Acawr+KzVfqA1H0FToqn46QL9Lr4nw== 10, 20 -- From: "Daniel Morrison" {dmorriso-at-stfx.ca} 10, 20 -- To: {Microscopy-at-microscopy.com} 10, 20 -- Content-Transfer-Encoding: 8bit 10, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6QCbJDk029410 ==============================End of - Headers==============================
Trilogy is a high pH antigen retrieval buffer used to unmask antigen sites in formalin fixed paraffin embedded tissue. It also has a detergent in it that can also deparaffinize tissue when used properly. I think it is an EDTA solution, pH 9-11?. As apposed to the normal Citrate buffer pH 6.0 that is normally used. Trilogy is only used when Citrate buffers and enzyme digestion, like trypsin, fail. We routinely used it in the clinical pathology lab on marker's like cd 10, cd 31, cd 4, cd 5, cd 8, etc.
"Effective unmasking, which reveals the antigen otherwise concealed by the negative effects of formalin fixation, has an undeniable and positive impact on the validation process. The mechanism of these negative effects is not yet completely understood but it is presumed that heating tissue sections in TrilogyTM causes the break up of the formalin cross-links that are believed to interfere with antigen-antibody binding. The discovery of the benefits of effective unmasking has been a revolutionary step in paving the road to standardization and the establishment of reliable validation procedures." From Cell Marque web site. They do not reveal their formula. =20
Hope this help's.
Rhonda Allen BA HT(ASCP)HTL, QIHC Histotechnology Specialist II Stowers Institute 1000 E. 50th Street Kansas City, MO 64110=20 816-926-4305
Rhonda Allen BA HT(ASCP)HTL, QIHC
==============================Original Headers============================== 13, 23 -- From rra-at-stowers-institute.org Wed Jul 26 09:44:20 2006 13, 23 -- Received: from stowers-institute.org (mail.stowers-institute.org [204.56.6.8]) 13, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QEiKkY009517 13, 23 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 09:44:20 -0500 13, 23 -- Received: from ([172.16.2.9]) 13, 23 -- by smtpkc01.stowers-institute.org with ESMTP id 4420336.1650497; 13, 23 -- Wed, 26 Jul 2006 09:43:50 -0500 13, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 23 -- Content-class: urn:content-classes:message 13, 23 -- MIME-Version: 1.0 13, 23 -- Content-Type: text/plain; 13, 23 -- charset="us-ascii" 13, 23 -- Subject: Trilogy 13, 23 -- Date: Wed, 26 Jul 2006 09:43:50 -0500 13, 23 -- Message-ID: {C28BAF593DC3314E9C0F3A50191C2E7804FB2300-at-EXCHKC03.stowers-institute.org} 13, 23 -- X-MS-Has-Attach: 13, 23 -- X-MS-TNEF-Correlator: 13, 23 -- Thread-Topic: Trilogy 13, 23 -- Thread-Index: AcawweiFHoky7NDoQ4yE/jGIQoPsPg== 13, 23 -- From: "Allen, Rhonda" {RRA-at-stowers-institute.org} 13, 23 -- To: {microscopy-at-microscopy.com} 13, 23 -- Content-Transfer-Encoding: 8bit 13, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6QEiKkY009517 ==============================End of - Headers==============================
Postdoc (or professional programmer with scientific background) – Portland State University (Portland, Oregon/USA)
Initially for one year at $35,000 (to $38,000 depending on experience and background) plus 50 % fringe benefits (health insurance, retirement benefits, etc.), available from January 2nd, 2007, onwards, extendable up to 3 years by mutual agreement and if continuing funding over ONAMI {http://www.onami.us/} earmarks (or alternative funding agencies) can be secured.
The Nanocrystallography Group {http://www.physics.pdx.edu/%7Epmoeck/research%20group} at the Department of Physics {http://www.physics.pdx.edu/} of Portland State University {http://www.pdx.edu/} (PSU) is seeking a (male/female) postdoc (or professional programmer with scientific background) for a project on Image-based nanocrystallography with database support: Lattice-fringe fingerprinting to identify unknown nanocrystal phases & Nanocrystal morphology from tilt protocols {http://www.physics.pdx.edu/%7Epmoeck/projects/ONAMI-nanometrology/white%20paper%202006.htm} .
Excellent computer skills (e.g. C++, JScript, Java, Python, Perl, PHP, and MySQL) are essential. A background in crystallography, mineralogy, or materials science and engineering at the Masters (or PhD level) is desirable. Advanced degrees in computer science may substitute.
PSU is with an estimated enrollment of about 30,000 students Oregon’s largest and only urban university. It is located in downtown {http://maps.google.com/maps?oi=map&q=Portland,+OR} Portland/Oregon {http://www.portlandonline.com/} , i.e. the metropolis of the so called “silicon forest”. Since high tech industries blend in with abundant natural beauty in this region and because of its moderate climate, Portland has consistently been ranked in the top ten of “America’s most livable cities”.
The search will be open until the position has been filled. Applications (CV, list of referees, reasons for coming to the US for applicants from aboard, list of publications if applicable, etc.) should be sent to:
Prof. Peter Moeck {http://www.physics.pdx.edu/%7Epmoeck/}
Applications by email to pmoeck-at-pdx.edu {mailto:pmoeck-at-pdx.edu} with attachments as *.pdf files are acceptable. Since there is a hyperlink at the title of the project above, prospective candidates are advised to deliver a statement on how they might contribute to this multi-institutional collaborative effort.
-- Peter Moeck (PhD, Dr. rer. nat.) Assistant Professor
Portland State University, Department of Physics P.O. Box 751, Portland, OR 97207-0751 Tel.: 503 725 4227, Departmental fax.: 503 725 9525
pmoeck-at-pdx.edu
(Office 404, Science Building 2, 1719 SW 10th Ave, Portland, OR 97201)
http:/www.physics.pdx.edu/~pmoeck
==============================Original Headers============================== 21, 20 -- From pmoeck-at-pdx.edu Wed Jul 26 10:50:57 2006 21, 20 -- Received: from njord.oit.pdx.edu (njord.oit.pdx.edu [131.252.120.57]) 21, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QFovaD020907 21, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 26 Jul 2006 10:50:57 -0500 21, 20 -- Received: from pdx.edu (71-34-86-40.ptld.qwest.net [71.34.86.40]) 21, 20 -- (authenticated bits=0) 21, 20 -- by njord.oit.pdx.edu (8.13.3+/8.13.1) with ESMTP id k6QFos7e028578 21, 20 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 21, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 26 Jul 2006 08:50:56 -0700 21, 20 -- X-Authentication-Warning: njord.oit.pdx.edu: Host 71-34-86-40.ptld.qwest.net [71.34.86.40] claimed to be pdx.edu 21, 20 -- Message-ID: {44C78F6C.4010904-at-pdx.edu} 21, 20 -- Date: Wed, 26 Jul 2006 08:51:08 -0700 21, 20 -- From: "Peter Moeck (Laptop - on the run)" {pmoeck-at-pdx.edu} 21, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 21, 20 -- X-Accept-Language: en-us, en 21, 20 -- MIME-Version: 1.0 21, 20 -- To: Microscopy-at-Microscopy.Com 21, 20 -- Subject: postdoc or professional programmer ($35k to $38k up to 3 years) 21, 20 -- Content-Type: text/plain; charset=windows-1252; format=flowed 21, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
MSc/PhD candidate positions in crystallography/applied physics
The Nanocrystallography Group {http://www.physics.pdx.edu/%7Epmoeck/research%20group} at the Department of Physics {http://www.physics.pdx.edu/} of Portland State University {http://www.pdx.edu/} is seeking candidates (m/f) for MSc and PhD projects in the field of transmission electron microscopy (TEM) based nanocrystallography. The graduate student projects focus on aspects of the development of novel nanocrystal characterization methods. These methods will utilize a combination of high-resolution phase contrast imaging in TEM and goniometry of direct lattice vectors {http://www.physics.pdx.edu/%7Epmoeck/goniometry.htm} .
The Nanocrystallography group is a user of Portland State University’s electron microscopy center {http://www.gatan.com/knowhow_8/psu_open_house.pdf} . Occasional travel to and work at the National Center for Electron Microscopy {http://ncem.lbl.gov/} , Lawrence Berkeley National Laboratory is anticipated to access some of the most advanced TEM instrumentation in the country. Summer research at the Department of Physics, Technical University Chemnitz may also become part of some projects.
The ideal candidate has a diploma of BSc. in crystallography, mineralogy, materials science and engineering, physics, or chemistry, and it interested in both geometrical-structural crystallography and transmission electron microscopy. Computer- and experimental skills are essential for the success of the projects. While some projects will require more experimental skills, some other projects will require more computer skills (notably JScript, Java, C++, Perl, PHP).
Portland State University is with an estimated enrollment of about 30,000 students Oregon’s largest and only urban university. Portland/Oregon {http://www.portlandonline.com/} is the major city in the so called “silicon forest”, i.e. a region where high tech industries blend in with abundant natural beauty, and has consistently been ranked in the top ten of “America’s most livable cities”. Information on student life in Portland {http://www.physics.pdx.edu/gep_files/gep.htm} can be found on our departmental web page. The search will be open until successful candidates have taken up their respective positions.
Applications (CV, references, etc.) should be sent to: Prof. Peter Moeck, Department of Physics, Portland State University, P.O. Box 751, Portland, Oregon 97207-0751
Michael has made an important point the type of DP heater is matched to the original fluid in the DP you cannot just change to another fluid, the fluid has to match the original fluid characteristics, boiling point for instance!
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {michael-at-shaffer.net} To: {protrain-at-emcourses.com} Sent: Tuesday, July 25, 2006 7:40 PM
Hello, I bought a microwave from panasonic with its inverter technology that lowers the wattage of the microwave.
I want to test it to see how well it can do processing for SEM. I was going to try bacteria on a surface, but anyone recommend other sensitive tissues? Leaves from plants maybe?
I'll post the results here if there is interest.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 4, 24 -- From gvrdolja-at-nature.berkeley.edu Wed Jul 26 17:44:36 2006 4, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QMia8U027109 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 17:44:36 -0500 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 3122BC1E33 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 15:44:36 -0700 (PDT) 4, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 4, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 4, 24 -- with ESMTP id 07939-08 for {microscopy-at-microscopy.com} ; 4, 24 -- Wed, 26 Jul 2006 15:44:29 -0700 (PDT) 4, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 4, 24 -- id 66CF3C1E1D; Wed, 26 Jul 2006 15:44:29 -0700 (PDT) 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 2ED48C1E33 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 15:44:28 -0700 (PDT) 4, 24 -- Date: Wed, 26 Jul 2006 15:44:27 -0700 (PDT) 4, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 4, 24 -- To: microscopy-at-microscopy.com 4, 24 -- Subject: microwave fixation 4, 24 -- Message-ID: {Pine.SOC.4.64.0607261542380.3594-at-nature.Berkeley.EDU} 4, 24 -- MIME-Version: 1.0 4, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
Debby Sherman and I thought it might be of interest for those using ultra-rapid freezing and freeze substitution approaches in their work to get together during the M&M 2006 meeting, in an informal setting. The notion is to share experiences (both good and bad) and ideas in using this methodology, that this will lead to improved success rate in sample preparation. If you are interested, please feel free to join us. The venue is the bar at the Holiday Inn at City Center, where we will have a large table set aside for a congenial setting. The time is Wednesday, August 2, beginning at 5:15 p.m. or so, and continuing until?
With best wishes for a successful M&M 2006,
Howard Berg
Director, Integrated Microscopy Facility Danforth Plant Science Center 975 N. Warson Rd. St. Louis, MO 63132
ph 314-587-1261 fx 314-587-1361 cell 314-378-2409 rhberg-at-danforthcenter.org www.danforthcenter.org visit this educational resource: http://www.danforthcenter.org/Cells/
==============================Original Headers============================== 12, 21 -- From RHBerg-at-danforthcenter.org Wed Jul 26 19:09:22 2006 12, 21 -- Received: from spm1.ddpsc.org (spm1.danforthcenter.org [65.254.111.26]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6R09Mpu006178 12, 21 -- for {Microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 19:09:22 -0500 12, 21 -- Received: from spm1.ddpsc.org (127.0.0.1) by spm1.ddpsc.org (MlfMTA v3.1r24) id hp02340171sm for {Microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 19:09:21 -0500 (envelope-from {RHBerg-at-danforthcenter.org} ) 12, 21 -- Received: from mail02.ddpsc.org ([10.101.0.23]) 12, 21 -- by spm1.ddpsc.org (SonicWALL 4.6.0.7527) 12, 21 -- with ESMTP; Wed, 26 Jul 2006 19:09:21 -0500 12, 21 -- Received: from [10.14.0.20] ([10.14.0.20] unverified) by mail02.ddpsc.org with Microsoft SMTPSVC(5.0.2195.6713); 12, 21 -- Wed, 26 Jul 2006 19:08:32 -0500 12, 21 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 21 -- Content-Transfer-Encoding: 7bit 12, 21 -- Message-Id: {13FDEFCB-A55A-4C64-85D8-07BD373A23D1-at-danforthcenter.org} 12, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 21 -- To: Microscopy-at-microscopy.com 12, 21 -- From: "R. Howard Berg" {rhberg-at-danforthcenter.org} 12, 21 -- Subject: Informal discussion, freeze substitution methodology, at M&M 2006 12, 21 -- Date: Wed, 26 Jul 2006 19:08:29 -0500 12, 21 -- X-Mailer: Apple Mail (2.750) 12, 21 -- X-OriginalArrivalTime: 27 Jul 2006 00:08:32.0040 (UTC) FILETIME=[CB9E9A80:01C6B110] 12, 21 -- X-Mlf-Version: 4.6.0.7527 ==============================End of - Headers==============================
I do believe that the comparatively warm water you are using in the cooling system would also be a factor. 25 degrees is fairly warm. I run my Denton cooling water at 15 degrees.
Ted Dunn The EMscope Co. Ld.
--- protrain-at-emcourses.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 } ---------------------------------------------------------------------------- } } Hi } } Michael has made an important point the type of DP } heater is matched to the } original fluid in the DP you cannot just change to } another fluid, the fluid } has to match the original fluid characteristics, } boiling point for instance! } } Steve Chapman } Senior Consultant Protrain } For electron microscopy consultancy and training } world wide } Tel +44 1280 816512 Fax +44 1280 814007 } Mobile +44 7711 606967 Web www.emcourses.com } } ----- Original Message ----- } X-from: {michael-at-shaffer.net} } To: {protrain-at-emcourses.com} } Sent: Tuesday, July 25, 2006 7:40 PM } Subject: [Microscopy] RE: [Microscopy}RE: Diff. Pump } Problem } } } } } } } } } } } ---------------------------------------------------------------------------- } } 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 } } } ---------------------------------------------------------------------------- } } } } WCB writes ... } } } } } As suggested by others, Stanton's problem with } his diffusion } } } pump may very well be due to insufficient heat } input from the } } } DP's heater. ... } } } } I remember a similar problem, which could have } well been the heater output } } ... But in this case it was because I had changed } the DP fluid to another } } that had a higher boiling pt. The heater did work } ... But it sure was } } sluggish in the beginning. } } } } Genuinely, Michael Shaffer :o) } } } } SEM/MLA Research Coordinator } } http://www.mun.ca/creait/maf/ } } Inco Innovation Centre } } Memorial University } } St. John's, NL } } } } } } } } ==============================Original } } Headers============================== } } 7, 21 -- From michael-at-shaffer.net Tue Jul 25 } 13:39:13 2006 } } 7, 21 -- Received: from ws6-5.us4.outblaze.com } (ws6-5.us4.outblaze.com } } [205.158.62.152]) } } 7, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } } k6PIdCN0016239 } } 7, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 25 } Jul 2006 13:39:12 -0500 } } 7, 21 -- Received: (qmail 24459 invoked from } network); 25 Jul 2006 } } 18:39:13 -0000 } } 7, 21 -- Received: from unknown (HELO rarewolf) } } (michael-at-shaffer.net-at-205.251.84.119) } } 7, 21 -- by ws6-5.us4.outblaze.com with SMTP; 25 } Jul 2006 18:39:12 -0000 } } 7, 21 -- From: "michael shaffer" } {michael-at-shaffer.net} } } 7, 21 -- To: "MSA Microscopy list" } {Microscopy-at-microscopy.com} } } 7, 21 -- Cc: {bigelow-at-engin.umich.edu} } } 7, 21 -- Subject: RE: [Microscopy] [Microscopy}RE: } Diff. Pump Problem } } 7, 21 -- Date: Tue, 25 Jul 2006 16:09:12 -0230 } } 7, 21 -- Message-ID: } {000b01c6b019$a08639e0$4701a8c0-at-rarewolf} } } 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 11 } } 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE } V6.00.2900.2869 } } 7, 21 -- Thread-Index: } AcawBILzsLXAAdW2Qhi/oRSyFMrupwAFIf3g } } 7, 21 -- In-Reply-To: } {200607251608.k6PG81A3025848-at-ns.microscopy.com} } } ==============================End of - } } Headers============================== } } } } } } } ==============================Original } Headers============================== } 7, 33 -- From protrain-at-emcourses.com Wed Jul 26 } 12:37:54 2006 } 7, 33 -- Received: from rd01.mail.x-isp.net } (rd01.mail.x-isp.net [213.253.171.197]) } 7, 33 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k6QHbrAv010973 } 7, 33 -- for {microscopy-at-microscopy.com} ; Wed, 26 } Jul 2006 12:37:53 -0500 } 7, 33 -- Received: from [62.69.65.166] } (helo=smtp-c.mail.legend.co.uk) } 7, 33 -- by rd01.mail.x-isp.net with smtp (Exim } 4.60) } 7, 33 -- (envelope-from {protrain-at-emcourses.com} ) } 7, 33 -- id 1G5nKS-00056m-GH } 7, 33 -- for microscopy-at-microscopy.com; Wed, 26 Jul } 2006 18:37:52 +0100 } 7, 33 -- Received: (qmail 10343 invoked from } network); 26 Jul 2006 17:37:50 -0000 } 7, 33 -- Received: from unknown (HELO advent) } (212.248.149.133) } 7, 33 -- by 0 with SMTP; 26 Jul 2006 17:37:50 } -0000 } 7, 33 -- Message-ID: } {001c01c6b0da$4b29d670$8595f8d4-at-advent} } 7, 33 -- Reply-To: "Steve Chapman" } {protrain-at-emcourses.com} } 7, 33 -- From: "Steve Chapman" } {protrain-at-emcourses.com} } 7, 33 -- To: {michael-at-shaffer.net} } 7, 33 -- Cc: "American Soc" } {microscopy-at-microscopy.com} } 7, 33 -- References: } {200607251840.k6PIeCt1018097-at-ns.microscopy.com} } 7, 33 -- Subject: Re: [Microscopy] RE: } [Microscopy}RE: Diff. Pump Problem } 7, 33 -- Date: Wed, 26 Jul 2006 18:37:59 +0100 } 7, 33 -- Organization: Protrain } 7, 33 -- MIME-Version: 1.0 } 7, 33 -- Content-Type: text/plain; } 7, 33 -- format=flowed; } 7, 33 -- charset="iso-8859-1"; } 7, 33 -- reply-type=original } 7, 33 -- Content-Transfer-Encoding: 7bit } 7, 33 -- X-Priority: 3 } 7, 33 -- X-MSMail-Priority: Normal } 7, 33 -- X-Mailer: Microsoft Outlook Express } 6.00.2900.2527 } 7, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE } V6.00.2900.2527 } 7, 33 -- X-tmReceived: from [62.69.65.166] } (helo=smtp-c.mail.legend.co.uk) } 7, 33 -- by rd01.mail.x-isp.net with smtp id } 1G5nKS-00056m-GH;Wed, 26 Jul 2006 18:37:52 +0100 } ==============================End of - } Headers============================== }
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The matter of changing from one type of diffusion pump fluid to another is a rather complicated one, which is discussed in some detail in Section 5.4.6 (p. 188) of my book 'Vacuum Methods in Electron Microscopy'. (Available from SPI, Ladd, M. E. Taylor, etc.) Theoretically, fluids which have similar slopes for their vapor pressure vs temperature curves, shown in Fig. 5.6 (p. 182), should have similar heats of vaporization, and therefore should require similar heat input from the DP heater, and so should be interchangeable. Note, however, that this plot involves logarithmic scales, so that small differences in slope may involve large differences in actual values. Therefore, just to be on the safe side, I would recommend checking with the pump's manufacturer before making a change. -- 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============================== 1, 14 -- From bigelow-at-engin.umich.edu Thu Jul 27 13:02:27 2006 1, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6RI2QD7029301 1, 14 -- for {microscopy-at-microscopy.com} ; Thu, 27 Jul 2006 13:02:27 -0500 1, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- by srvr22.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k6RI2PDa022846 1, 14 -- for {microscopy-at-microscopy.com} ; Thu, 27 Jul 2006 14:02:25 -0400 (EDT) 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06210200c0eeabb2e36e-at-[141.212.131.221]} 1, 14 -- Date: Thu, 27 Jul 2006 14:02:24 -0400 1, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 1, 14 -- Subject: [Microscopy] RE: Changing DP fluids 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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Email: dwaugh-at-kent.edu Name: David Waugh
Organization: Kent State University
Title-Subject: [Filtered] SEM mechanical stage parts (Amray 1645)
Question: We have an Amray 1645 with a mechanical stage that has just died. The treaded shaft that actuates the Y axis of the stage has sheared off at the location of the shear pin that holds the gear on the non-threaded part of the shaft. See links for pictures of broken shaft. Does anyone have some spare parts that that they could "part" with? Many Thanks, David
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Question: We are using 6nm-10 beads conjugated to IgG (or IgM) antibodies to detect IgG (or IgM) Monoclonal antibodies bound to a viral surface antigens. Does anyone know the distance(s) from the antigens to the beads?
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Email: camiller-at-anatomy.iupui Name: Caroline Miller
Organization: Indiana University
Title-Subject: [Filtered] FIG on Cryomicroscopy
Question: I would like to let everyone know that a FIG has been approved for Cryomicroscopy. There will be an organizational meeting Tuesday, Aug. 1st from 12:00-12:30PM in Room 308 in Festival Hall during the MM2006 Meeting. I encourage everyone who is interested to attend this meeting. Your input will help to determine the direction of this new FIG. Maybe this is a more formal setting, but still a good way for all us to have a venue for discussing problems and sucesses with high pressure freezing, automatic freeze substitution and cryo ultra microtomy.
Try Fjeld, Inc. A lot of stages were and are still made by them.
gary g.
At 03:10 PM 7/27/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Your question is not so easy to answer. Probably only an approximation can be given. One has to consider at least: * dimensions of antibodies (often described as approximately 10nm max dimension for an IgG) * dimensions of particles (6 or 10 nm in this case) * orientation of antibodies on particle surfaces
If you would consider the first antibody only, sitting on the antigen, and realizing the antibody has a number of hinges in its structure, this would mean that in a worst case assumption assuming full flexibility around the antigen the primary antibody is located within a circle with a radius of about 10nm with the antigen binding site as the geometrical center. The radius will be bigger when working with IgM. Gold conjugates are often imagined as a particle sitting on the Fc- tail of a secondary antibody. Some secondaries may actually be oriented that way, but it is very likely that some secondaries are oriented side-on on the gold particle surface. So there may very well be a range of conjugate sizes if you like. Again, a worst case assumption would probably bring the center of a 10nm particle of a secondary conjugate about 15 nm away (for an IgG conjugate) from the binding site on the primary.
Add all this up and the worst case assumption amounts to an area of probability with a radius of 25nm within which the centre of the gold particle should be located. That is what the situation may be as long as the specimen is in a buffer. During drying the dimensions of the whole structure may change while collapsing onto the specimen. Is it reaonable to assume worst case conditions? From our own work in the past, even three step labeling involving an unlabelled secondary antibody and protein A gold conjugate to label the outer segment of a membrane protein in bacteria still showed the majority of the particles located outside the cells and not projected over the periplasmic space or even the outer membrane.
What has often made me wonder in working with viruses or Virus Like Particles (VLP) and immunolabelling is that the gold label is often surrounding the virus particles rather than actually covering the structures.
In practice, and theoretically for resolution reasons, it may be worthwhile to reduce the labelling to a one step one, with the gold particle sitting directly on the primary antibody, or even on an Fab fragment of the primary. One step labelling of course also has its disadvantage in that the presence of the gold particle on the primary antibody may result in steric hindrance. Using an ultra small colloidal particle or covalently coupled particle and post-labelling enhancement would be the best choice to reduce those risks.
Well, I am not sure if all this helps, please feel free to get in touch if you would like to discuss this further.
Cheers
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz
David Waugh wrote: ========================================================== Question: We have an Amray 1645 with a mechanical stage that has just died. The treaded shaft that actuates the Y axis of the stage has sheared off at the location of the shear pin that holds the gear on the non-threaded part of the shaft. See links for pictures of broken shaft. Does anyone have some spare parts that that they could "part" with? Many Thanks, David =============================================== Contact
Mr. Clark Houghton Secondary Images 2371 Emery Lane Winchester, OH 45697
They are a "third party" service firm and are almost a neighbor of yours. They specialize in servicing AMRAY SEMs. I think they could help you with this problem.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
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Email: susan.vanhorn-at-sunysb.edu Name: Susan Van Horn
Organization: SUNY -at- StonyBrook
Title-Subject: [Filtered] glow discharge
Question: I am getting mixed information concerning glow discharged grids: once they are glow discharged how long does it last??? and how is it accomplished?? thanks sue
On Jul 28, 2006, at 2:15 PM, susan.vanhorn-at-sunysb.edu wrote:
} Question: I am getting mixed information concerning glow discharged } grids: once they are glow discharged how long does it last??? and how } is it accomplished?? } Dear Sue, The effect of glow-discharging--to make the carbon hydrophyllic--fades over time, and how long it lasts depends on just how hydrophyllic the carbon must be to satisfy your particular need. For cryoEM, we have found that putting the material on the grid for freezing is best done within a few minutes after a 2 minute glow-discharge step. However, for other specimens a longer interval is OK, and for bacteriorhodopsin, the carbon surface should be "aged" over a weekend for best results. An additional complication is that the conversion of the surface from hydrophyllic to hydrophobic depends on the environmental conditions, so it can vary from lab to lab. It is accomplished by the bombardment of the carbon by oxygen or nitrogen ions (or whatever else is in the plasma); in order to make extremely hydrophyllic grids, amyl amine has been introduced into the glow-discharge volume, so in that case I think that molecular ions or fragments from the amyl amine must be particularly effective, but I don't know which component(s) is the actual active one. 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 Fri Jul 28 16:47:42 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 k6SLlgX2020748 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 28 Jul 2006 16:47:42 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP 4, 22 -- id C0B4C3545D; Fri, 28 Jul 2006 14:47:41 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP 4, 22 -- id 255C92EF86; Fri, 28 Jul 2006 14:47:36 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200607282115.k6SLFq72000454-at-ns.microscopy.com} 4, 22 -- References: {200607282115.k6SLFq72000454-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {285b9ef1b3a605900886a7a66441976c-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: glow discharge 4, 22 -- Date: Fri, 28 Jul 2006 14:49:15 -0700 4, 22 -- To: susan.vanhorn-at-sunysb.edu, microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
I am working on a MSc project in which I am studying the effect of microwave exposure to water during freezing, in the context of electron microscopy sample preparation.
A couple of papers published indicate a reduction in the size of ice crystals when water (including that in biological samples) is frozen in the presence of microwaves (references below).
There is even a commercially available instrument, the Eiko MF-2 (see Eiko Instruments' website at http://www.kagaku.com/eiko/e-products/ electron/mf_2.html), which that company claims provides a reduction in ice crystal size when freezing (I have no commercial interest whatsoever in this company, by the way).
I would be very pleased to hear from anyone who has studied this effect (even if the work was unpublished) or knows of any other work on the subject. It would also be interesting to hear from anyone who has actually used the MF-2!
Best regards
Richard
References: Hanyu, Y., Ichikawa, M. and Matsumoto, G: An improved cryofixation method: cryoquenching of small tissue blocks during microwave irradiation. Journal of Microscopy, 165:Pt 2 255-271. February 1992
Jackson, T.H., Ungan, U., Critser, J.K. and Gao, D: Novel microwave technology for cryopreservation of biomaterials by suppression of apparent ice formation. Cryobiology 34, 363-372. 1997
Richard Easingwood Otago Centre for Electron Microscopy c/- Department of Anatomy & Structural Biology University of Otago, Dunedin, New Zealand ph: 0064 3 479 7301 cell: 021 222 4759 fax: 0064 3 479 7254 or 479 5086 http://ocem.otago.ac.nz
==============================Original Headers============================== 14, 20 -- From richard.easingwood-at-stonebow.otago.ac.nz Sat Jul 29 17:23:30 2006 14, 20 -- Received: from mailhub1.otago.ac.nz (mailhub1.otago.ac.nz [139.80.64.218]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6TMNTPS003051 14, 20 -- for {Microscopy-at-microscopy.com} ; Sat, 29 Jul 2006 17:23:29 -0500 14, 20 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 14, 20 -- by mailhub1.otago.ac.nz (8.13.6/8.13.6) with ESMTP id k6TMNSVs019262 14, 20 -- for {Microscopy-at-microscopy.com} ; Sun, 30 Jul 2006 10:23:28 +1200 14, 20 -- Received: from dialin093096.otago.ac.nz ([139.80.93.96]) 14, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 14, 20 -- id 1G6xDT-0001cs-7x 14, 20 -- for Microscopy-at-microscopy.com; Sun, 30 Jul 2006 10:23:27 +1200 14, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 14, 20 -- Content-Transfer-Encoding: 7bit 14, 20 -- Message-Id: {27F38189-5C17-48CD-9925-D5BEE529E611-at-stonebow.otago.ac.nz} 14, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 14, 20 -- To: Microscopy-at-microscopy.com 14, 20 -- From: Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} 14, 20 -- Subject: Microwave assisted freezing 14, 20 -- Date: Sun, 30 Jul 2006 10:23:25 +1200 14, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Susan Van Horn wrote: ==================================================== Question: I am getting mixed information concerning glow discharged=20 grids: once they are glow discharged how long does it last??? and how is it accomplished?? thanks ====================================================== We have found two different approaches can "work":
a) If the glow discharge is done in a small table top plasma etcher, such as the SPI Plasma Prep II, see URL http://www.2spi.com/catalog/instruments/etchers1.shtml, using either an "air" or nitrogen plasma, carbon coated grids can be made hydrophilic and they will maintain their hydrophilic nature for 30-60 days before they start to lose these enhanced properties. The "mixed information" you reference could be due to the fact that there are a number of different systems that produce a "glow discharge". The SPI system operates at 100 watts. Systems of lower power seem to produce grids that will retain the enhanced properties for a shorter period of time.
b) Evaporation of a molecular layer of Victawet, see URL http://www.2spi.com/catalog/spec_prep/evapor_3c.shtml This is a phosphate based water soluble surfactant so it could possibly interfere with at least some experiments. But if you can get away with using it, it will leave the grids with their enhanced properties essentially "forever".
Disclaimer: SPI Supplies offers both the SPI Plasma Prep II Plasma Etcher and also, Victawet, so we have a vested interest in seeing more people using both products.
Chuck ================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President=20 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ====================================================
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I don't know the best stain for your polymer. However, I know that osmium tetroxide will stain any unsaturation in your material and, very importantly, will probably stain little else in the molecule. Specifically, OsO4 adds across the C=C bond. By comparison, ruthenium has much less specificity since it will stain the C=C bond and other hydrocarbon components of the molecule.
Staining can, and probably should, be done in the vapor phase. Affix your specimen to the inside of the cap to a vial containing OsO4 solution and stain overnight. Degas for several hours. Remember that OsO4 is toxic and all work should be done in a good fume hood.
Regards,
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
kmoulton-at-usm.m aine.edu To gary.m.brown-at-exxonmobil.com 07/28/06 04:19 cc PM Subject [Microscopy] viaWWW: TEM staining Please respond methods to kmoulton-at-usm.m aine.edu
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Samsung Austin Semiconductor is currently looking for a TEM Engineer to join its Materials & Analysis Team. The M&A team is responsible for surface analysis and fab inspection. The surface analysis team will work primarily with SIMS, Auger analysis and FIB and TEM imaging. The fab inspection team will focus primarily on bevel and edge defects and tool level defect fingerprinting.
Job Duties
* Manage TEM Microscope * Perform TEM imaging and summary write ups * Oversee TEM Sample prep and provide feedback * Work with unit process and product technology engineers to resolve device and in line issues
Requirements
* BS in EE, ChemE, or Material Science * TEM microscope operation * DRAM device knowledge * FLASH device knowledge
Please send resumes or questions to Alison Rao at arao-at-sas.samsung.com
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==============================Original Headers============================== 14, 26 -- From arao-at-SAS.Samsung.com Mon Jul 31 16:58:25 2006 14, 26 -- Received: from worldsecure.samsungaustin.com (pix-080.sas.samsung.com [207.207.40.80]) 14, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6VLwPZb001521 14, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 31 Jul 2006 16:58:25 -0500 14, 26 -- Received: from 105.192.16.223 by worldsecure.samsungaustin.com with 14, 26 -- ESMTP (Samsung Austin Semiconductor SMTP Relay - No UCE Please! (Email 14, 26 -- Firewall v6.2.0)); Mon, 31 Jul 2006 16:58:14 -0500 14, 26 -- X-Server-Uuid: 8F66684A-E1C5-43A6-96BB-810E9723A306 14, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 14, 26 -- Content-class: urn:content-classes:message 14, 26 -- MIME-Version: 1.0 14, 26 -- Subject: TEM- TEM Engineer Job Opening at Samsung Austin Semiconductor 14, 26 -- Date: Mon, 31 Jul 2006 16:58:14 -0500 14, 26 -- Message-ID: {3E231818A261CF418B23994A39B064370355034F-at-exchange} 14, 26 -- X-MS-Has-Attach: 14, 26 -- X-MS-TNEF-Correlator: 14, 26 -- Thread-Topic: TEM- TEM Engineer Job Opening at Samsung Austin 14, 26 -- Semiconductor 14, 26 -- Thread-Index: Aca07GvFA1/6fjUhRkGAHucFCTYdqw== 14, 26 -- From: "Alison Rao - SAS(HR)" {arao-at-SAS.Samsung.com} 14, 26 -- To: Microscopy-at-Microscopy.Com 14, 26 -- X-WSS-ID: 68D0A37C2LK2363061-01-01 14, 26 -- Content-Type: text/plain; 14, 26 -- charset=us-ascii 14, 26 -- Content-Transfer-Encoding: 8bit 14, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6VLwPZb001521 ==============================End of - Headers==============================
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