There is an intriguing new technology which is under development that might solve this problem. NT-MDT has integrated an AFM with an ultramicrotome (Leica UC6-NT), called NTegra Tomo. The developer, Dr. Anton Efimov, has recently been experimenting with a cryo version. Tomo has proven very helpful in imaging and elucidating 3D nanostructures for things like dried emulsions and biological entities (c. elegans). If the nanoemulsion is cryo-stable, the cryo version of Tomo should be a good solution. Also, because the AFM uses local differences in elasticity to image different phases, there would be no need to stain.
Please contact me off-line if you would like to have Dr. Efimov try your investigator's sample as part of his test program.
Hope this was helpful,
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through July. Call us today for details.
P. S. Need a good general reference or light microscopy text for the Spring semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.
At 10:10 AM 10/27/2006, dsoren-at-umich.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 16, 17 -- From bfoster-at-mme1.com Thu Feb 1 07:47:35 2007 16, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 16, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l11DlYO7026957 16, 17 -- for {microscopy-at-microscopy.com} ; Thu, 1 Feb 2007 07:47:35 -0600 16, 17 -- Received: (qmail 24989 invoked by uid 2020); 1 Feb 2007 08:15:08 -0600 16, 17 -- Received: from 207.47.25.42.static.nextweb.net (HELO barbsd505.mme1.com) (207.47.25.42) 16, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 1 Feb 2007 08:15:08 -0600 16, 17 -- Message-Id: {7.0.1.0.0.20070131125629.01d8e860-at-mme1.com} 16, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 16, 17 -- Date: Wed, 31 Jan 2007 13:03:28 -0600 16, 17 -- To: dsoren-at-umich.edu, microscopy-at-microscopy.com 16, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 16, 17 -- Subject: Re: [Microscopy] EM nanoemulsion 16, 17 -- In-Reply-To: {200610271333.k9RDXVUb006433-at-ns.microscopy.com} 16, 17 -- References: {200610271333.k9RDXVUb006433-at-ns.microscopy.com} 16, 17 -- Mime-Version: 1.0 16, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Thanks to everyone who responded to my maleic acid buffer query. We now have some things to try.
By the way, the following response is irresistable, courtesy of the incomparable Snoop Leunissen (Jan, that is). Foshizzle, microscopists rock, dog.
Enjoy.
Randy
"Buffer Rap
For anyone who likes to do EM buffers at times are a hell of a wham the chemistry lacks in every way yet we like to have a good display
So what is this all about mall ee 8? Does that stuff accommodate uranyl acetate? at what pH, what ionic strength? I better ask the LIST for some reference
With some advice here and a helping hand I am sure I can pretend I understand So I take some stuff from the lab supply Mix it together and hope I will get by
Wow, man, what happens, it's workin' alright! The negative stain is clear and bright! No precipitate, the structures they are fine It works! Now I can advice the next in line.
Anyway, for mallE8 to work alright, you see you need two solutions, mark them A and B Empty bottle (C) in the middle now that should do And mixing left and right will be the clue
Solution A has sodium hydrogen maleate 23.2 grams if it's trihydrate Dissolve in 200 ml 1M Sodium Hydroxide And make to 1 liter with distilled water alright
Solution B is simple just point 1 Molar NaO-age Solution C is the trick, Now don't get into a rage!
Chorus: Take 25 mls out of bottle A transfer to Bottle C without further delay Mix in x mls of B, top up to 100 cc Get approximate pH from the listing you will see
pH x ml 0.1 M NaOH
5.2 7.2 5.4 10.5 5.6 15.3 5.8 20.8 6.0 26.9
For Tris maleate it is much the same Two stocks again, it's almost lame The first holds Tris as well as Maleic acid 24.2 and 23.2 grams, yep that's it!
And before I forget, make a liter of that now the other stock again is just NaO-age a plain 0.2 Molar is all that it takes
Chorus...
pH x ml 0.2 M NaOH
5.4 5.4 5.6 7.75 5.8 10.25 6.0 13.0
--------------
Apologies, the listings don't rhyme. Disclaimer: I will not be responsible for anyone getting hurt while trying to rap the numbers!
X-from: "Data for Biochemical research", by Dawson, Elliot, Elliot and Jones Clarendon Press, Oxford, 3rd ed.
Recipes for maleate buffer (J.Am.Chem.Soc 51 (1929), 1754) and Tris/maleate buffer (PSEBM 68 (1948) p354 or Meth Enzym. 1 (1955) 138"
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 28, 23 -- From TindallR-at-missouri.edu Thu Feb 1 08:42:34 2007 28, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 28, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l11EgYFF006727 28, 23 -- for {microscopy-at-microscopy.com} ; Thu, 1 Feb 2007 08:42:34 -0600 28, 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); 28, 23 -- Thu, 1 Feb 2007 08:42:33 -0600 28, 23 -- x-mimeole: Produced By Microsoft Exchange V6.5 28, 23 -- Content-class: urn:content-classes:message 28, 23 -- MIME-Version: 1.0 28, 23 -- Content-Type: text/plain; 28, 23 -- charset="us-ascii" 28, 23 -- Subject: Maleate vs. maleic vs. malic, YO 28, 23 -- Date: Thu, 1 Feb 2007 08:42:33 -0600 28, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68DA0-at-UM-XMAIL08.um.umsystem.edu} 28, 23 -- X-MS-Has-Attach: 28, 23 -- X-MS-TNEF-Correlator: 28, 23 -- Thread-Topic: Maleate vs. maleic vs. malic, YO 28, 23 -- Thread-Index: AcdGDzdmACxBEBI2RqSd73T4Ih9kDw== 28, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 28, 23 -- To: {microscopy-at-microscopy.com} 28, 23 -- X-OriginalArrivalTime: 01 Feb 2007 14:42:33.0631 (UTC) FILETIME=[35511AF0:01C7460F] 28, 23 -- Content-Transfer-Encoding: 8bit 28, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l11EgYFF006727 ==============================End of - Headers==============================
Since the list played a big part in my decision to go through with LASIK 2.5 years ago, I'll put in 2 cents on this topic as well.
My doctor did warn about floaters, halos, possible mis-correction as side effects and then gave me a realistic assessment of what it would mean should I suffer these side effects. He spent a great deal of time addressing my concerns and assured me that such problems were quite rare and often very minor.
In the 2 months following the surgery, I experienced halos and starburst patterns around streetlights and headlights while driving at night. Eventually these symptoms subsided and I see less starburst-type patterns now than I ever did before the surgery. At 2.5 years post surgery, I now have a slight floater in my right eye which I rarely notice unless conditions are just so. I never noticed it at all until 4-5 months ago. That said, I know several friends and family members who have floaters that have NEVER had LASIK surgery, so I do not feel confident that LASIK was the cause.
All in all, the improvement in my vision from about 20:250 to 20:20 has been an immensely positive development. I would recommend LASIK to anyone who has been declared a good candidate by reputable eye surgeon. Do your research on the physician. I had no less than 4 recommendations from optometrists with no connection to each other. I also spoke with 3 patients he had treated in the past, so I felt pretty comfortable that he was competent.
For anyone, especially a microscopist, your vision isn't something to bargain shop for. One place in town offers LASIK "as low as $599 per eye". It didn't take long to find out that they have higher complication rates. Ultimately, I paid close to $3,000 for both eyes and was confident that I was getting the best treatment available in the area.
Cheers, Jay
On 1/29/07, Jessica.Wagner-at-childrens.harvard.edu {Jessica.Wagner-at-childrens.harvard.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 } ---------------------------------------------------------------------------- } } John, } } I missed that thread, but I'll be happy to add my two cents now. } Assuming the PVD (posterior vitreous detachment) doesn't get any worse, } for me the benefits of LASIK still outweigh this minor negative. The } 'floater' that I see is an annoyance, but it doesn't truly inhibit my } vision through the scope (or outside the scope for that matter). } } But I had LASIK only a year ago, and was never warned PVD could be a } complication, so this says to me that there is still much unknown about } the procedure and it's results. Unfortunately, complications of laser } eye surgeries really aren't tracked all that well; doctors are not } required to report them, unless they are related to a device, but even } then, many doctors are not aware of FDA regulations about device event } reporting or how/to whom they should report adverse events. } } Here is an article about PVD's and LASIK: } http://www.springerlink.com/content/j3100858467pu5k1/ } } And thanks to everyone who offered imaging advice! } } Jessica } } } -----Original Message----- } X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] } Sent: Monday, January 29, 2007 12:56 PM } To: Wagner, Jessica } Subject: [Microscopy] RE: imaging the occular view through another port? } } } } } } ------------------------------------------------------------------------ } ---- } 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 } ------------------------------------------------------------------------ } ---- } } Jessica; } Some time ago, there was a thread about LASIK and microscopists. } I do not recall any of the reports mentioning this sort of post-surgical } vision difficulty. Is this perhaps a subject that should be re-visited? } } John Mardinly } Intel } } Disclaimer: The opinions of this author do not represent the opinions of } Intel Corporation. } } -----Original Message----- } X-from: Jessica.Wagner-at-childrens.harvard.edu } [mailto:Jessica.Wagner-at-childrens.harvard.edu] } Sent: Friday, January 26, 2007 12:25 PM } To: Mardinly, John } Subject: [Microscopy] imaging the occular view through another port? } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } Hello list. I have a question that's just for fun. I'm using a Nikon } TE-2000 wth all 4 ports and a beam splitter that can direct light 50/50 } split between two ports. Is there a way to set up a camera to image not } a sample but specifically the image that I'm seeing? I have a minor } defect in my eye, a wrinkle caused by slight detachment of the vitreous } (I think due to having LASIK done). When I look through the scope at a } bright field, I can see an image of the wrinkle. I'm curious to know if } I can capture it, but my guess is that the image only exists in the } occulars? Maybe I could somehow use a dichroic mirror? } } Thanks, } Jessica } } } } ==============================Original } Headers============================== } 4, 37 -- From Jessica.Wagner-at-childrens.harvard.edu Fri Jan 26 14:24:29 } 2007 4, 37 -- Received: from mail2.childrenshospital.org } (mail2.childrenshospital.org [134.174.20.64]) } 4, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l0QKOTAx003925 } 4, 37 -- for {Microscopy-at-microscopy.com} ; Fri, 26 Jan 2007 } 14:24:29 -0600 } 4, 37 -- Received: from tumsmtp1.CHBOSTON.ORG (tumsmtp1.tch.harvard.edu } [10.1.101.46]) } 4, 37 -- by mail2.childrenshospital.org (Postfix) with ESMTP id } B770480F1 } 4, 37 -- for {Microscopy-at-microscopy.com} ; Fri, 26 Jan 2007 } 15:24:26 -0500 (EST) } 4, 37 -- Received: from 10.1.102.175 by tumsmtp1.CHBOSTON.ORG with ESMTP } (MMS 4, 37 -- SMTP Relay (Email Firewall v6.3.0)); Fri, 26 Jan 2007 } 15:24:13 -0500 4, 37 -- X-Server-Uuid: } 1F337096-A893-456A-BE4C-C6341410F3EE } 4, 37 -- Received: from CHEXV4.CHBOSTON.ORG ([10.1.102.188]) by 4, 37 -- } chexsmtp2.CHBOSTON.ORG with Microsoft SMTPSVC(6.0.3790.1830); Fri, 26 4, } 37 -- Jan 2007 15:24:13 -0500 4, 37 -- X-MimeOLE: Produced By Microsoft } Exchange V6.5 4, 37 -- Content-class: urn:content-classes:message 4, 37 } -- MIME-Version: 1.0 4, 37 -- Subject: imaging the occular view through } another port? 4, 37 -- Date: Fri, 26 Jan 2007 15:24:12 -0500 4, 37 -- } Message-ID: {6B335BE3804E0A40978750E51EFC076108B83B-at-CHEXV4.CHBOSTON.ORG} } 4, 37 -- X-MS-Has-Attach: } 4, 37 -- X-MS-TNEF-Correlator: } 4, 37 -- Thread-Topic: imaging the occular view through another port? 4, } 37 -- Thread-Index: AcdBh/FJTzzdmMO7TlybuNWpOytJvw== 4, 37 -- From: } "Wagner, Jessica" {Jessica.Wagner-at-childrens.harvard.edu} } 4, 37 -- To: Microscopy-at-microscopy.com } 4, 37 -- X-OriginalArrivalTime: 26 Jan 2007 20:24:13.0614 (UTC) 4, 37 -- } FILETIME=[F1C7C4E0:01C74187] 4, 37 -- X-TMWD-Spam-Summary: } TS=20070126202415; SEV=2.2.0; DFV=B2007012608; 4, 37 -- } IFV=2.0.4,4.0-9; AIF=B2007012608; RPD=5.02.0004; ENG=IBF; 4, 37 -- } RPDID=7374723D303030312E30413031303230332E34354241363336462E303032442C73 } 733D312C6667733D30; } 4, 37 -- CAT=NONE; CON=NONE } 4, 37 -- X-MMS-Spam-Filter-ID: B2007012608_5.02.0004_4.0-9 } 4, 37 -- X-WSS-ID: 69A4BCE73B47225184-01-01 } 4, 37 -- Content-Type: text/plain; } 4, 37 -- charset=iso-8859-1 } 4, 37 -- Content-Transfer-Encoding: 8bit } 4, 37 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id l0QKOTAx003925 ==============================End of } - Headers============================== } } } ==============================Original } Headers============================== } 13, 34 -- From john.mardinly-at-intel.com Mon Jan 29 11:51:39 2007 13, 34 } -- Received: from mga02.intel.com (mga02.intel.com [134.134.136.20]) } 13, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l0THpd49013923 } 13, 34 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 29 Jan 2007 } 11:51:39 -0600 } 13, 34 -- Received: from orsmga001.jf.intel.com ([10.7.209.18]) } 13, 34 -- by mga02.intel.com with ESMTP; 29 Jan 2007 09:51:39 -0800 } 13, 34 -- Received: from fmsmsx333.amr.corp.intel.com ([132.233.42.2]) } 13, 34 -- by orsmga001.jf.intel.com with ESMTP; 29 Jan 2007 09:51:38 } -0800 } 13, 34 -- X-ExtLoop1: 1 } 13, 34 -- X-IronPort-AV: i="4.13,253,1167638400"; } 13, 34 -- d="scan'208"; a="190293831:sNHT77980763" } 13, 34 -- Received: from scsmsx411.amr.corp.intel.com ([10.3.90.30]) by } fmsmsx333.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); } 13, 34 -- Mon, 29 Jan 2007 09:51:31 -0800 } 13, 34 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by } scsmsx411.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); } 13, 34 -- Mon, 29 Jan 2007 09:51:30 -0800 } 13, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 13, 34 -- Content-class: urn:content-classes:message } 13, 34 -- MIME-Version: 1.0 } 13, 34 -- Content-Type: text/plain; } 13, 34 -- charset="us-ascii" } 13, 34 -- Subject: RE: [Microscopy] imaging the occular view through } another port? 13, 34 -- Date: Mon, 29 Jan 2007 09:51:29 -0800 13, 34 -- } Message-ID: } {1DF4C4D62339DB4C9C98DF04213995B60263F0B5-at-scsmsx413.amr.corp.intel.com} } 13, 34 -- In-Reply-To: {200701262024.l0QKObkm004099-at-ns.microscopy.com} } 13, 34 -- X-MS-Has-Attach: } 13, 34 -- X-MS-TNEF-Correlator: } 13, 34 -- Thread-Topic: [Microscopy] imaging the occular view through } another port? 13, 34 -- Thread-Index: } AcdBiAPf8S8hIZO1T+u/pIAfVESIzACRKOmg } 13, 34 -- From: "Mardinly, John" {john.mardinly-at-intel.com} } 13, 34 -- To: {Jessica.Wagner-at-childrens.harvard.edu} } 13, 34 -- Cc: {Microscopy-at-msa.microscopy.com} } 13, 34 -- X-OriginalArrivalTime: 29 Jan 2007 17:51:30.0799 (UTC) } FILETIME=[1B8E67F0:01C743CE] 13, 34 -- Content-Transfer-Encoding: 8bit } 13, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id l0THpd49013923 ==============================End of } - Headers============================== } } } } ==============================Original Headers============================== } 28, 37 -- From Jessica.Wagner-at-childrens.harvard.edu Mon Jan 29 16:18:54 2007 } 28, 37 -- Received: from mail2.childrenshospital.org (mail2.childrenshospital.org [134.174.20.64]) } 28, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l0TMIrbi030608 } 28, 37 -- for {Microscopy-at-microscopy.com} ; Mon, 29 Jan 2007 16:18:53 -0600 } 28, 37 -- Received: from tumsmtp1.CHBOSTON.ORG (tumsmtp1.tch.harvard.edu [10.1.101.46]) } 28, 37 -- by mail2.childrenshospital.org (Postfix) with ESMTP id C3D458082 } 28, 37 -- for {Microscopy-at-microscopy.com} ; Mon, 29 Jan 2007 17:18:50 -0500 (EST) } 28, 37 -- Received: from 10.1.102.174 by tumsmtp1.CHBOSTON.ORG with ESMTP (MMS } 28, 37 -- SMTP Relay (Email Firewall v6.3.0)); Mon, 29 Jan 2007 17:18:43 -0500 } 28, 37 -- X-Server-Uuid: 1F337096-A893-456A-BE4C-C6341410F3EE } 28, 37 -- Received: from CHEXV4.CHBOSTON.ORG ([10.1.102.188]) by } 28, 37 -- chexsmtp1.CHBOSTON.ORG with Microsoft SMTPSVC(6.0.3790.1830); Mon, 29 } 28, 37 -- Jan 2007 17:18:43 -0500 } 28, 37 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 28, 37 -- Content-class: urn:content-classes:message } 28, 37 -- MIME-Version: 1.0 } 28, 37 -- Subject: LASIK and microscopists } 28, 37 -- Date: Mon, 29 Jan 2007 17:18:42 -0500 } 28, 37 -- Message-ID: {6B335BE3804E0A40978750E51EFC076107BF91-at-CHEXV4.CHBOSTON.ORG} } 28, 37 -- X-MS-Has-Attach: } 28, 37 -- X-MS-TNEF-Correlator: } 28, 37 -- Thread-Topic: LASIK and microscopists } 28, 37 -- Thread-Index: AcdD829TWE3SHiDVT4OxJPpkd6QQAg== } 28, 37 -- From: "Wagner, Jessica" {Jessica.Wagner-at-childrens.harvard.edu} } 28, 37 -- To: Microscopy-at-microscopy.com } 28, 37 -- X-OriginalArrivalTime: 29 Jan 2007 22:18:43.0039 (UTC) } 28, 37 -- FILETIME=[6F8416F0:01C743F3] } 28, 37 -- X-TMWD-Spam-Summary: TS=20070129221845; SEV=2.2.0; DFV=B2007012910; } 28, 37 -- IFV=2.0.4,4.0-9; AIF=B2007012910; RPD=5.02.0004; ENG=IBF; } 28, 37 -- RPDID=7374723D303030312E30413031303230312E34354245373243342E303046372C73733D312C6667733D30; } 28, 37 -- CAT=NONE; CON=NONE } 28, 37 -- X-MMS-Spam-Filter-ID: B2007012910_5.02.0004_4.0-9 } 28, 37 -- X-WSS-ID: 69A0AD493B47707430-01-01 } 28, 37 -- Content-Type: text/plain; } 28, 37 -- charset=us-ascii } 28, 37 -- Content-Transfer-Encoding: 8bit } 28, 37 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l0TMIrbi030608 } ==============================End of - Headers============================== }
==============================Original Headers============================== 7, 24 -- From microtomy-at-gmail.com Thu Feb 1 13:40:01 2007 7, 24 -- Received: from an-out-0708.google.com (an-out-0708.google.com [209.85.132.241]) 7, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l11Jdxdv025805 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 1 Feb 2007 13:40:00 -0600 7, 24 -- Received: by an-out-0708.google.com with SMTP id b20so410486ana 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 01 Feb 2007 11:39:58 -0800 (PST) 7, 24 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 7, 24 -- d=gmail.com; s=beta; 7, 24 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 7, 24 -- b=F/llkUFBGYvVSS1zkZCPM3qAorSUKjDoNZjyuyFOWZbRNdLsT32eS+DRkjL2M8wztuOU0xFA5pd5iRelzZl0/NIj8k0noBYjnUOHY4sLgtS1uzzDRTw8vfzLGUScmm7EIjkAJG+QnNmN4lnMTG7mVRBfuS/AexO924ROUqbbYYE= 7, 24 -- Received: by 10.114.25.3 with SMTP id 3mr191988way.1170358797597; 7, 24 -- Thu, 01 Feb 2007 11:39:57 -0800 (PST) 7, 24 -- Received: by 10.114.149.12 with HTTP; Thu, 1 Feb 2007 11:39:57 -0800 (PST) 7, 24 -- Message-ID: {ef6bda5c0702011139l43448b9bnad0ebfee04a0c617-at-mail.gmail.com} 7, 24 -- Date: Thu, 1 Feb 2007 13:39:57 -0600 7, 24 -- From: "Jay Campbell" {microtomy-at-gmail.com} 7, 24 -- To: Microscopy-at-microscopy.com 7, 24 -- Subject: Re: [Microscopy] LASIK and microscopists 7, 24 -- In-Reply-To: {200701292221.l0TMLQwM000624-at-ns.microscopy.com} 7, 24 -- MIME-Version: 1.0 7, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 24 -- Content-Transfer-Encoding: 7bit 7, 24 -- Content-Disposition: inline 7, 24 -- References: {200701292221.l0TMLQwM000624-at-ns.microscopy.com} ==============================End of - Headers==============================
This isn't about LASIK, but I caught the bit about floaters and just want to relay my experience, if only to prevent others from going through the ordeal I did last summer.
Don't ignore floaters, even if they've been noticeable for a long time. My right eye always had a significant number of them, and my optometrist told me to come back if I noticed an increased number of them. That's not very easy quantify over time, but in hindsight the number probably did increase in the months before I had a partial detachment of the retina. No other symptoms until a ominous black spot appeared in the corner of my vision. The surgeon said that the whole thing could have fallen off at any time, probably resulting in total blindness in that eye. Fortunately surgery corrected everything, and six months later I have nearly perfect (well, as perfect as it was before) vision again in that eye.
Get your eyes dilated and checked for retinal tears *every* time you have an eye exam, especially if you're over 40. I know, it's a pain, but losing binocular vision is a much bigger pain! Retinal detachment is *not* most common in boxers, drag racers, sky divers - people who get their heads banged around a lot. It happens most often to people who are strongly nearsighted. I know quite a few of those in the microscopy world and I don't think any of them would be keen on saving operating expenses by only buying monocular scopes.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I can also add that the number of eye 'floaters' I have is increasing with age (I'm 43), and I have never had eye surgery. The thought of trying to zap them, and nothing else important, with a laser is terrifying, but it is getting more difficult to use my light microscope. I am also very nearsighted and wear contact lenses; does anyone know if there's a correlation?
Thanks, Jane
Jane L. LaGoy Laboratory Services Manager/ Development Engineer Bodycote North America 155 River Street Andover, MA 01810 978-470-1620 x450 FAX: 978-475-2951 jane.lagoy-at-bodycote.com The only people to get even with are those who have helped you.
==============================Original Headers============================== 7, 29 -- From Jane.LaGoy-at-bodycote.com Thu Feb 1 15:38:25 2007 7, 29 -- Received: from outbound8-blu-R.bigfish.com (outbound-blu.frontbridge.com [65.55.251.16]) 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l11LcPg4018321 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 15:38:25 -0600 7, 29 -- Received: from outbound8-blu.bigfish.com (localhost.localdomain [127.0.0.1]) 7, 29 -- by outbound8-blu-R.bigfish.com (Postfix) with ESMTP id D0CAB9DBA9F 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:24 +0000 (UTC) 7, 29 -- Received: from mail172-blu-R.bigfish.com (unknown [10.1.252.3]) 7, 29 -- by outbound8-blu.bigfish.com (Postfix) with ESMTP id BC80360004F 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:24 +0000 (UTC) 7, 29 -- Received: from mail172-blu (localhost.localdomain [127.0.0.1]) 7, 29 -- by mail172-blu-R.bigfish.com (Postfix) with ESMTP id 705B71A305DF 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:24 +0000 (UTC) 7, 29 -- X-BigFish: VP 7, 29 -- Received: by mail172-blu (MessageSwitch) id 1170365904156892_23649; Thu, 1 Feb 2007 21:38:24 +0000 (UCT) 7, 29 -- Received: from Exchange.BODYCOTE-IMT.COM (mail.bodycote-imt.com [65.249.143.82]) 7, 29 -- by mail172-blu.bigfish.com (Postfix) with ESMTP id 9222A890067 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:23 +0000 (UTC) 7, 29 -- Received: by mail.bodycote-imt.com with Internet Mail Service (5.5.2657.72) 7, 29 -- id {D0XTGK8G} ; Thu, 1 Feb 2007 16:44:11 -0500 7, 29 -- Message-ID: {CBB9714FDC67D411B39400D0B73C4B73028A4905-at-mail.bodycote-imt.com} 7, 29 -- From: Jane LaGoy {Jane.LaGoy-at-bodycote.com} 7, 29 -- To: "Microscopy Listserver (E-mail)" {Microscopy-at-MSA.Microscopy.com} 7, 29 -- Subject: eye floaters 7, 29 -- Date: Thu, 1 Feb 2007 16:44:11 -0500 7, 29 -- MIME-Version: 1.0 7, 29 -- X-Mailer: Internet Mail Service (5.5.2657.72) 7, 29 -- Content-Type: text/plain; 7, 29 -- charset="iso-8859-1" ==============================End of - Headers==============================
I'm also extremely nearsighted and my floaters have increased with age. Sometimes when I'm reading, I have to move my move head to get a pesky one out of the way. I had them before wearing hard contacts for 12 years and I still have them. So I don't think there's a correlation but I realize my opinion is not a scientific study. My optometrist once explained to me why very near-sighted people should have their retinas examined frequently for tears/detachments. Near-sighted eyeballs are longer front to back than normal eyeballs. He said one can be born with normal-sized retinas stretched to fit the larger eyeballs. (My father is near-sighted; my mother is not.) These 'stretched' retinas are more prone to tears/damage/detachments than normal ones. I have no training in eye physiology so I was taking him at his word. As microscopists, our eyes are more valuable than our hands, which are pretty darn valuable. Take care of them.
Jane.LaGoy-at-bodycote.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I can also add that the number of eye 'floaters' I have is increasing with } age (I'm 43), and I have never had eye surgery. The thought of trying to } zap them, and nothing else important, with a laser is terrifying, but it is } getting more difficult to use my light microscope. I am also very } nearsighted and wear contact lenses; does anyone know if there's a } correlation? } } Thanks, Jane } } Jane L. LaGoy } Laboratory Services Manager/ } Development Engineer } Bodycote North America } 155 River Street } Andover, MA 01810 } 978-470-1620 x450 } FAX: 978-475-2951 } jane.lagoy-at-bodycote.com } The only people to get even with are those who have helped you. } } } } } } ==============================Original Headers============================== } 7, 29 -- From Jane.LaGoy-at-bodycote.com Thu Feb 1 15:38:25 2007 } 7, 29 -- Received: from outbound8-blu-R.bigfish.com (outbound-blu.frontbridge.com [65.55.251.16]) } 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l11LcPg4018321 } 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 15:38:25 -0600 } 7, 29 -- Received: from outbound8-blu.bigfish.com (localhost.localdomain [127.0.0.1]) } 7, 29 -- by outbound8-blu-R.bigfish.com (Postfix) with ESMTP id D0CAB9DBA9F } 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:24 +0000 (UTC) } 7, 29 -- Received: from mail172-blu-R.bigfish.com (unknown [10.1.252.3]) } 7, 29 -- by outbound8-blu.bigfish.com (Postfix) with ESMTP id BC80360004F } 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:24 +0000 (UTC) } 7, 29 -- Received: from mail172-blu (localhost.localdomain [127.0.0.1]) } 7, 29 -- by mail172-blu-R.bigfish.com (Postfix) with ESMTP id 705B71A305DF } 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:24 +0000 (UTC) } 7, 29 -- X-BigFish: VP } 7, 29 -- Received: by mail172-blu (MessageSwitch) id 1170365904156892_23649; Thu, 1 Feb 2007 21:38:24 +0000 (UCT) } 7, 29 -- Received: from Exchange.BODYCOTE-IMT.COM (mail.bodycote-imt.com [65.249.143.82]) } 7, 29 -- by mail172-blu.bigfish.com (Postfix) with ESMTP id 9222A890067 } 7, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2007 21:38:23 +0000 (UTC) } 7, 29 -- Received: by mail.bodycote-imt.com with Internet Mail Service (5.5.2657.72) } 7, 29 -- id {D0XTGK8G} ; Thu, 1 Feb 2007 16:44:11 -0500 } 7, 29 -- Message-ID: {CBB9714FDC67D411B39400D0B73C4B73028A4905-at-mail.bodycote-imt.com} } 7, 29 -- From: Jane LaGoy {Jane.LaGoy-at-bodycote.com} } 7, 29 -- To: "Microscopy Listserver (E-mail)" {Microscopy-at-MSA.Microscopy.com} } 7, 29 -- Subject: eye floaters } 7, 29 -- Date: Thu, 1 Feb 2007 16:44:11 -0500 } 7, 29 -- MIME-Version: 1.0 } 7, 29 -- X-Mailer: Internet Mail Service (5.5.2657.72) } 7, 29 -- Content-Type: text/plain; } 7, 29 -- charset="iso-8859-1" } ==============================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, 23 -- From r-holdford-at-ti.com Thu Feb 1 17:53:30 2007 4, 23 -- Received: from bear.ext.ti.com (bear.ext.ti.com [192.94.94.41]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l11NrTNd031781 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 1 Feb 2007 17:53:29 -0600 4, 23 -- Received: from dlep32.itg.ti.com ([157.170.170.70]) 4, 23 -- by bear.ext.ti.com (8.13.7/8.13.7) with ESMTP id l11NrOJ4002155 4, 23 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 1 Feb 2007 17:53:29 -0600 4, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 23 -- by dlep32.itg.ti.com (8.13.7/8.13.7) with ESMTP id l11NrNd9002807 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 1 Feb 2007 17:53:23 -0600 (CST) 4, 23 -- Message-ID: {45C27D73.7070207-at-ti.com} 4, 23 -- Date: Thu, 01 Feb 2007 17:53:23 -0600 4, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 23 -- Organization: SC Packaging Development -- FA Development 4, 23 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) 4, 23 -- MIME-Version: 1.0 4, 23 -- To: MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 23 -- Subject: Re: [Microscopy] eye floaters 4, 23 -- References: {200702012138.l11Lcdl7018633-at-ns.microscopy.com} 4, 23 -- In-Reply-To: {200702012138.l11Lcdl7018633-at-ns.microscopy.com} 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am about to prepare some zebra fish heads for SEM - to look at morphology & take some measurements. What do you think are the pros & cons of critical point drying versus HMDS drying after fixation in GDA+PVP in SCB followed by osmium+potassium ferrocyanide followed by staining in aqueous UA before dehydration for small fish heads? I have had success with the HMDS drying using cultured cells and insect tissue. The thermocirculator attached to my CPD has problems - it is difficult to heat slowly enough so until I can replace it I thought HMDS might be an alternative method.
Many thanks Ursula ---------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
==============================Original Headers============================== 5, 24 -- From U.J.Potter-at-bath.ac.uk Fri Feb 2 10:22:01 2007 5, 24 -- Received: from kelly.bath.ac.uk (kelly.bath.ac.uk [138.38.32.20]) 5, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12GM0BQ030415 5, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 10:22:01 -0600 5, 24 -- Received: from amos.bath.ac.uk ([138.38.32.36] ident=mmdf) 5, 24 -- by kelly.bath.ac.uk with smtp 5, 24 -- (envelope-from {U.J.Potter-at-bath.ac.uk} ) 5, 24 -- id 1HD1Al-00084h-4z 5, 24 -- for Microscopy-at-microscopy.com; Fri, 02 Feb 2007 16:22:00 +0000 5, 24 -- Received: from eapc-03.campus.bath.ac.uk 5, 24 -- ( eapc-03.campus.bath.ac.uk [138.38.136.63] ) by bath.ac.uk 5, 24 -- id aa02496 for {Microscopy-at-microscopy.com} ; 2 Feb 2007 16:22 +0000 5, 24 -- Date: Fri, 02 Feb 2007 16:21:58 +0000 5, 24 -- From: Ursula Potter {U.J.Potter-at-bath.ac.uk} 5, 24 -- To: Microscopy-at-microscopy.com 5, 24 -- Subject: SEM of Zebra fish 5, 24 -- Message-ID: {26073250.1170433318-at-eapc-03.campus.bath.ac.uk} 5, 24 -- Originator-Info: login-id=mssujp; server=imaphost.bath.ac.uk 5, 24 -- X-Mailer: Mulberry/3.1.0 (Win32) 5, 24 -- MIME-Version: 1.0 5, 24 -- Content-Type: text/plain; charset=us-ascii; format=flowed 5, 24 -- Content-Transfer-Encoding: 7bit 5, 24 -- Content-Disposition: inline 5, 24 -- X-Scanner: 6ca9c34fc388b0ebba8f13ee036c7c05e14a6226 ==============================End of - Headers==============================
If anyone is interested in an AMRAY 1600 the University of Missouri-Columbia has just listed it on Ebay. Go to : www.surplus.missouri.edu then the eBay items link.
Any questions feel free to contact me. Lou Ross Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=2227 email: rosslm-at-missouri.edu http://www.emc.missouri.edu
==============================Original Headers============================== 4, 21 -- From RossLM-at-missouri.edu Fri Feb 2 10:44:05 2007 4, 21 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12Gi5GW009736 4, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 10:44:05 -0600 4, 21 -- Received: from UM-XMAIL06.um.umsystem.edu ([209.106.228.32]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 4, 21 -- Fri, 2 Feb 2007 10:44:04 -0600 4, 21 -- Received: from 128.206.78.120 ([128.206.78.120]) by UM-XMAIL06.um.umsystem.edu ([209.106.228.42]) via Exchange Front-End Server webmail.um.umsystem.edu ([209.106.228.21]) with Microsoft Exchange Server HTTP-DAV ; 4, 21 -- Fri, 2 Feb 2007 16:44:05 +0000 4, 21 -- User-Agent: Microsoft-Entourage/11.3.3.061214 4, 21 -- Date: Fri, 02 Feb 2007 10:44:08 -0600 4, 21 -- Subject: AMRAY 1600 on EBAY 4, 21 -- From: Lou Ross {rosslm-at-missouri.edu} 4, 21 -- To: {Microscopy-at-Microscopy.Com} 4, 21 -- Message-ID: {C1E8C678.86FF%rosslm-at-missouri.edu} 4, 21 -- Thread-Topic: AMRAY 1600 on EBAY 4, 21 -- Thread-Index: AcdG6VuDmiHmqrLcEduCuQAKlX496A== 4, 21 -- Mime-version: 1.0 4, 21 -- Content-type: text/plain; 4, 21 -- charset="US-ASCII" 4, 21 -- Content-transfer-encoding: 7bit 4, 21 -- X-OriginalArrivalTime: 02 Feb 2007 16:44:05.0089 (UTC) FILETIME=[59C70D10:01C746E9] ==============================End of - Headers==============================
A colleague needs to SEM image cells growing in monolayer on a coverslip in a thin layer of collagen matrix. I tried to prepare it more or less usual way - aldehyde fixation in cacodylate (2% formaldehyde/2% glutaraldehyde for about 90 min) followed by osmium, dehydration in increasing concentration of ethanol, replacing ethanol with amyl acetate and eventually CPD from carbon dioxide. The results are suboptimal at best - cells seem to shring and get extracted. Does anybody have a good idea they would like to share?
Thanks,
Michal
==============================Original Headers============================== 4, 17 -- From Michal.Jarnik-at-fccc.edu Fri Feb 2 13:55:35 2007 4, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12JtY1x025400 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 13:55:34 -0600 4, 17 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) 4, 17 -- (authenticated bits=0) 4, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id l12JtYEb022147 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 14:55:34 -0500 (EST) 4, 17 -- Message-ID: {45C39735.3020002-at-fccc.edu} 4, 17 -- Date: Fri, 02 Feb 2007 14:55:33 -0500 4, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} 4, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) 4, 17 -- MIME-Version: 1.0 4, 17 -- To: Microscopy-at-Microscopy.Com 4, 17 -- Subject: SEM of cells in monolayer 4, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Thanks to all who responded to my inquiry; I see an opthamologist annually but have not asked about this problem before, so I certainly will now.
Jane L. LaGoy Laboratory Services Manager/ Development Engineer Bodycote North America 155 River Street Andover, MA 01810 978-470-1620 x450 FAX: 978-475-2951 jane.lagoy-at-bodycote.com The only people to get even with are those who have helped you.
==============================Original Headers============================== 6, 29 -- From Jane.LaGoy-at-bodycote.com Fri Feb 2 14:02:25 2007 6, 29 -- Received: from outbound6-blu-R.bigfish.com (outbound-blu.frontbridge.com [65.55.251.16]) 6, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12K2PA1032739 6, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 2 Feb 2007 14:02:25 -0600 6, 29 -- Received: from outbound6-blu.bigfish.com (localhost.localdomain [127.0.0.1]) 6, 29 -- by outbound6-blu-R.bigfish.com (Postfix) with ESMTP id AB4F4108DC4C 6, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 2 Feb 2007 20:02:24 +0000 (UTC) 6, 29 -- Received: from mail32-blu-R.bigfish.com (unknown [10.1.252.3]) 6, 29 -- by outbound6-blu.bigfish.com (Postfix) with ESMTP id 9FA99BE805A 6, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 2 Feb 2007 20:02:24 +0000 (UTC) 6, 29 -- Received: from mail32-blu (localhost.localdomain [127.0.0.1]) 6, 29 -- by mail32-blu-R.bigfish.com (Postfix) with ESMTP id 54A7111B8213 6, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 2 Feb 2007 20:02:24 +0000 (UTC) 6, 29 -- X-BigFish: VP 6, 29 -- Received: by mail32-blu (MessageSwitch) id 1170446543486657_20339; Fri, 2 Feb 2007 20:02:23 +0000 (UCT) 6, 29 -- Received: from Exchange.BODYCOTE-IMT.COM (mail.bodycote-imt.com [65.249.143.82]) 6, 29 -- by mail32-blu.bigfish.com (Postfix) with ESMTP id C55831B5009E 6, 29 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 2 Feb 2007 20:02:22 +0000 (UTC) 6, 29 -- Received: by mail.bodycote-imt.com with Internet Mail Service (5.5.2657.72) 6, 29 -- id {D0XTGL7S} ; Fri, 2 Feb 2007 15:08:11 -0500 6, 29 -- Message-ID: {CBB9714FDC67D411B39400D0B73C4B73028A491B-at-mail.bodycote-imt.com} 6, 29 -- From: Jane LaGoy {Jane.LaGoy-at-bodycote.com} 6, 29 -- To: "Microscopy Listserver (E-mail)" {Microscopy-at-MSA.Microscopy.com} 6, 29 -- Subject: eye floater info - thanks! 6, 29 -- Date: Fri, 2 Feb 2007 15:08:00 -0500 6, 29 -- MIME-Version: 1.0 6, 29 -- X-Mailer: Internet Mail Service (5.5.2657.72) 6, 29 -- Content-Type: text/plain; 6, 29 -- charset="iso-8859-1" ==============================End of - Headers==============================
First, skip the amyl acetate, it's not needed. Second, how many soak-purge cycles did you do in the CPD? I.e., Fill the chamber, flush with lqCO2 until the EtOH is gone, let cells soak X minutes, then purge with lqCO2, and repeat N times. I found a monolayer on coversilps usually only needed 3 soaks for 5 minutes each, but sometimes could require 4 or 5 soaks. If all of the EtOH (or amyl acetate) is not removed, you will get serious shrinkage.
I also didn't bother with formaldehye or osmium, just 1.25% glut (2% is pretty strong). Depending on the kV you're using, 1% OsO4 may be helpful, though. Fixation in OsO4 should only need an hour. Glut can go 1 to 2 hours.
What % EtOH did you start the dehydration at? I've used 30% and 50% successfully, any higher is too high, and sometimes one needs to start at 15%. How long in each EtOH step? 5 minutes should be enough.
And then, you will get some shrinkage no matter what you do.
Other things to try: Hexamethyldisilizane (HMDS). After EtOH dehydration, go through a 2:1, 1:2 EtOH:HMDS, 3 X 100% HMDS series and then air-dry for 1 to 2 hours. Room temp. or at 60 deg. C, sometimes one works better than the other. Do in a hood!
If you have the proper equipment, freeze-drying can work very well, and since there are no chemical fixatives or dehydrating agents involved the cells may be "more real".
Phil
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==============================Original Headers============================== 9, 22 -- From oshel1pe-at-cmich.edu Fri Feb 2 14:45:10 2007 9, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 9, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12KjAg2016170 9, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 14:45:10 -0600 9, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 9, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l12LBn0K017601 9, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 16:12:17 -0500 9, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 9, 22 -- Fri, 2 Feb 2007 15:45:07 -0500 9, 22 -- Mime-Version: 1.0 9, 22 -- Message-Id: {f0623090dc1e95043ad68-at-[141.209.160.249]} 9, 22 -- In-Reply-To: {200702022002.l12K2LWK032597-at-ns.microscopy.com} 9, 22 -- References: {200702022002.l12K2LWK032597-at-ns.microscopy.com} 9, 22 -- Date: Fri, 2 Feb 2007 15:45:06 -0500 9, 22 -- To: Microscopy-at-microscopy.com 9, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 9, 22 -- Subject: Re: [Microscopy] SEM of cells in monolayer 9, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 9, 22 -- X-OriginalArrivalTime: 02 Feb 2007 20:45:07.0491 (UTC) FILETIME=[060A7730:01C7470B] 9, 22 -- X-CanItPRO-Stream: default 9, 22 -- X-Spam-Score: -3.4 () J_CHICKENPOX_44,L_EXCH_MF 9, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
The protocol sounds very "usual" as you say, and I wonder if something just went wrong along the way?
You didn't mention any osmotic agents in the fixative, sometimes beneficial for cultured cells; various things and concentrations are used. I just did some samples that used 7.5% sucrose in the fixatives and buffer washes until the OsO4 was rinsed out.
A somewhat longer time in the glutaraldehyde for SEM is sometimes beneficial; the cells toughen a bit - tip from our old Polaron CPD manual...
Also, while I don't think it is the problem, it is not necessary to use amyl acetate: using 100% ethanol works just fine for the CPD process. Make sure the ethanol is really dry - store ethanol for final changes over good molecular sieves - Type 3A, recently baked, ~5% of ethanol volume, let is stand some days well sealed, don't stir up fines. The CO2 also needs to be a dry grade with good molecular sieve trap on the line.
Make sure to exchane well to get all the ethanol out. The cells on coverglass should exchange quickly, but were they in some "capsule" that limited exchange? Was the vapor phase clear when it went supercritical, or hazy? Was there any smell of amyl acetate when you opened the chanmber?
Could the coverglasses have gone dry at some point - even in the CPD process? Did you do this job yourself, or have an underpaid, un-benefitted work-study student do the work? Sorry, that's not fair; I used to be one; they aren't ALL inattentive.... The liquid drops as gas phase pressure increases; maybe they got above the liquid surface? The instructions printed on our Balzers CPD-030 unit say to "drain and refill several times" but the sample should never really be drained from liquid during the flushing exchanges; always do partial changes to keep the sample submerged.
Hope this helps.
Dale Callaham Central Microscopy Facility c/o Microbiology Department Morrill 4 N, Rm.1 University of Massachusetts Amherst, MA 01003
http://www.bio.umass.edu/microscopy
Michal.Jarnik-at-fccc.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 } ---------------------------------------------------------------------------- } } Dear Listers, } } A colleague needs to SEM image cells growing in monolayer on a coverslip } in a thin layer of collagen matrix. I tried to prepare it more or less } usual way - aldehyde fixation in cacodylate (2% formaldehyde/2% } glutaraldehyde for about 90 min) followed by osmium, dehydration in } increasing concentration of ethanol, replacing ethanol with amyl acetate } and eventually CPD from carbon dioxide. The results are suboptimal at } best - cells seem to shring and get extracted. Does anybody have a good } idea they would like to share? } } Thanks, } } Michal } } ==============================Original Headers============================== } 4, 17 -- From Michal.Jarnik-at-fccc.edu Fri Feb 2 13:55:35 2007 } 4, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) } 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12JtY1x025400 } 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 13:55:34 -0600 } 4, 17 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) } 4, 17 -- (authenticated bits=0) } 4, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id l12JtYEb022147 } 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 14:55:34 -0500 (EST) } 4, 17 -- Message-ID: {45C39735.3020002-at-fccc.edu} } 4, 17 -- Date: Fri, 02 Feb 2007 14:55:33 -0500 } 4, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} } 4, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) } 4, 17 -- MIME-Version: 1.0 } 4, 17 -- To: Microscopy-at-Microscopy.Com } 4, 17 -- Subject: SEM of cells in monolayer } 4, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 17 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers==============================
==============================Original Headers============================== 12, 21 -- From dac-at-research.umass.edu Fri Feb 2 15:07:18 2007 12, 21 -- Received: from race3.oit.umass.edu (race3.oit.umass.edu [128.119.101.39]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12L7IlV027654 12, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 15:07:18 -0600 12, 21 -- Received: from [172.30.55.164] (eutopia.bio.umass.edu [128.119.55.30]) 12, 21 -- (authenticated bits=0) 12, 21 -- by race3.oit.umass.edu (8.13.7/8.13.7) with ESMTP id l12L6usu002749 12, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 12, 21 -- Fri, 2 Feb 2007 16:06:56 -0500 12, 21 -- Message-ID: {45C3A833.6020607-at-research.umass.edu} 12, 21 -- Date: Fri, 02 Feb 2007 16:08:03 -0500 12, 21 -- From: Dale Callaham {dac-at-research.umass.edu} 12, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.8.1.2pre) Gecko/20070111 SeaMonkey/1.1 12, 21 -- MIME-Version: 1.0 12, 21 -- To: Michal.Jarnik-at-fccc.edu, Microscopy Listserver {Microscopy-at-microscopy.com} 12, 21 -- Subject: Re: [Microscopy] SEM of cells in monolayer 12, 21 -- References: {200702022005.l12K5e5r010545-at-ns.microscopy.com} 12, 21 -- In-Reply-To: {200702022005.l12K5e5r010545-at-ns.microscopy.com} 12, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 21 -- Content-Transfer-Encoding: 7bit 12, 21 -- X-Whitelist: TRUE ==============================End of - Headers==============================
Greetings Bea, As an aging graduate of the associates degree program of the Madison Area Technical College (MATC), I have read this thread with particular interest. All the comments to date have been consistent with my experience. I entered the program a couple of years prior to your age; another classmate was a couple years older than you are now. We both have done well. I haven't found age to be an impediment. I think your life experience, coupled with the recent training that some employers find attractive, will make you a good prospect if you do well in the program (as we all know you will!).
There are many opportunities that will be open to you with just the associates degree, simply because these programs are well known in the small world of the microscopy community for providing people who can walk into a lab with the demonstrated unique little fiddly, intensely controlled skills that we require to do our work. It is hard to know if someone coming in off the street can do that stuff, even if they have more relevant academic training. It is also true that there are opportunities that will be closed to you, as another commenter observed.
The best action you can take to minimize the effect of your non-science background is to be prepared to take on additional training to customize your fit with whatever job you find. Microscopy contains many areas of specialization, so it is likely that any new job will require some extra training, for you or anyone. If you do well in the program, and project your willingness to jump right back in with additional classes as a new-hire (often this can be done on the employer's nickel), this combined with your life experience will be attractive to many employers.
Having said that, I had to do the same thing and after 2 years of school and a baby in the family it was a painful prospect! But it did make all the difference in the new job. After five and a half years in the field I feel very much a part of the microscopy community, and love the work. I hope you decide to join us, Bea! Sincerely, Matt
-----Original Message----- X-from: beadrysdale-at-yahoo.com [mailto:beadrysdale-at-yahoo.com] Sent: Wednesday, January 31, 2007 9:38 AM To: stephenson-at-impactanalytical.com
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Title-Subject: [Filtered] Job outlook for microscopy technicians
Question: Hello. I am considering undertaking a two-year associate degree program to become a microscopy technician. I was wondering if anyone would be willing to comment on the job prospects I might have as new graduate with no science experience beyond this associate degree. The college's placement stats. look good but I would like a second opinion. Also, I am in my early forties and wonder if that would work against me finding a job (one of the draws of microscopy is that it is such a specialized skill that I would think employers would look beyond age). Thanks for any help you can give me.
We often run into the same problem. I think some occurs when cells are left with minimal fluid for even very short times while dehydrating. Surface tension is a real problem with very little fluid coverage when changes are being made. It is best to leave a little fluid in the culture dish and do a few more changes rather than risk the problems of excess shrinkage. Even under the best of conditions, some shrinkage is inevitable.
Years ago I did a large number of cell cultures using ducupan resin to infiltrate the cells. Cells were fixed and then infiltrated with successive mixtures of Ducupan:H2O. The excess 100% resin was washed away using propylene oxide and then cultures polymerized. This minimized shrinkage but also you should expect that surface detail could also be obscured if any resin remained on the outside of the cells.
This method did minimize breakage of long processes from axonal and dendritic cells.
Debby
Debby Sherman, Director 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
} From: {Michal.Jarnik-at-fccc.edu} } Reply-To: {Michal.Jarnik-at-fccc.edu} } Date: Fri, 2 Feb 2007 13:59:27 -0600 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] SEM of cells in monolayer } } } } } ---------------------------------------------------------------------------- } 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, } } A colleague needs to SEM image cells growing in monolayer on a coverslip } in a thin layer of collagen matrix. I tried to prepare it more or less } usual way - aldehyde fixation in cacodylate (2% formaldehyde/2% } glutaraldehyde for about 90 min) followed by osmium, dehydration in } increasing concentration of ethanol, replacing ethanol with amyl acetate } and eventually CPD from carbon dioxide. The results are suboptimal at } best - cells seem to shring and get extracted. Does anybody have a good } idea they would like to share? } } Thanks, } } Michal } } ==============================Original Headers============================== } 4, 17 -- From Michal.Jarnik-at-fccc.edu Fri Feb 2 13:55:35 2007 } 4, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) } 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } l12JtY1x025400 } 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 13:55:34 -0600 } 4, 17 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) } 4, 17 -- (authenticated bits=0) } 4, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id l12JtYEb022147 } 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 14:55:34 -0500 } (EST) } 4, 17 -- Message-ID: {45C39735.3020002-at-fccc.edu} } 4, 17 -- Date: Fri, 02 Feb 2007 14:55:33 -0500 } 4, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} } 4, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) } 4, 17 -- MIME-Version: 1.0 } 4, 17 -- To: Microscopy-at-Microscopy.Com } 4, 17 -- Subject: SEM of cells in monolayer } 4, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 17 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 23 -- From dsherman-at-purdue.edu Fri Feb 2 16:28:36 2007 9, 23 -- Received: from 1061exfe02.adpc.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12MSa2o019001 9, 23 -- for {microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 16:28:36 -0600 9, 23 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe02.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 9, 23 -- Fri, 2 Feb 2007 17:28:36 -0500 9, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) 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 ; 9, 23 -- Fri, 2 Feb 2007 22:28:36 +0000 9, 23 -- User-Agent: Microsoft-Entourage/11.3.3.061214 9, 23 -- Date: Fri, 02 Feb 2007 17:28:35 -0500 9, 23 -- Subject: Re: [Microscopy] SEM of cells in monolayer 9, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 9, 23 -- To: {Michal.Jarnik-at-fccc.edu} , 9, 23 -- "message to: MSA list" {microscopy-at-microscopy.com} 9, 23 -- Message-ID: {C1E92543.18A02%dsherman-at-purdue.edu} 9, 23 -- Thread-Topic: [Microscopy] SEM of cells in monolayer 9, 23 -- Thread-Index: AcdHGXoBuG18erMMEdu8XQARJN08Mg== 9, 23 -- In-Reply-To: {200702021959.l12JxRoe029306-at-ns.microscopy.com} 9, 23 -- Mime-version: 1.0 9, 23 -- Content-type: text/plain; 9, 23 -- charset="US-ASCII" 9, 23 -- Content-transfer-encoding: 7bit 9, 23 -- X-OriginalArrivalTime: 02 Feb 2007 22:28:36.0656 (UTC) FILETIME=[7AFDDB00:01C74719] ==============================End of - Headers==============================
When I was working with cultured fibroblast monolayers for SEM back in the 70's, I used aldehydes and osmium, as most do, and experienced ripped cells and ones that appeared to 'crack' along the surface, particularly where long filopodia extended from the cell bodies. But, my TEM looked fine. So for kicks, I tried my usual TEM protocol for the SEM samples, which used the same aldehydes and osmium but also used uranyl acetate as a post-fix. I had excellent results. No cracks or tears at all.
Something you might wish to try.
Ann Hein Lehman Assistant Director, Electron Microscopy Facility Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
-----Original Message----- X-from: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu] Sent: Friday, February 02, 2007 5:32 PM To: Lehman, Ann R
We often run into the same problem. I think some occurs when cells are left with minimal fluid for even very short times while dehydrating. Surface tension is a real problem with very little fluid coverage when changes are being made. It is best to leave a little fluid in the culture dish and do a few more changes rather than risk the problems of excess shrinkage. Even under the best of conditions, some shrinkage is inevitable.
Years ago I did a large number of cell cultures using ducupan resin to infiltrate the cells. Cells were fixed and then infiltrated with successive mixtures of Ducupan:H2O. The excess 100% resin was washed away using propylene oxide and then cultures polymerized. This minimized shrinkage but also you should expect that surface detail could also be obscured if any resin remained on the outside of the cells.
This method did minimize breakage of long processes from axonal and dendritic cells.
Debby
Debby Sherman, Director 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
} From: {Michal.Jarnik-at-fccc.edu} } Reply-To: {Michal.Jarnik-at-fccc.edu} } Date: Fri, 2 Feb 2007 13:59:27 -0600 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] SEM of cells in monolayer } } } } } ------------------------------------------------------------------------ ---- } 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, } } A colleague needs to SEM image cells growing in monolayer on a coverslip } in a thin layer of collagen matrix. I tried to prepare it more or less } usual way - aldehyde fixation in cacodylate (2% formaldehyde/2% } glutaraldehyde for about 90 min) followed by osmium, dehydration in } increasing concentration of ethanol, replacing ethanol with amyl acetate } and eventually CPD from carbon dioxide. The results are suboptimal at } best - cells seem to shring and get extracted. Does anybody have a good } idea they would like to share? } } Thanks, } } Michal } } ==============================Original Headers============================== } 4, 17 -- From Michal.Jarnik-at-fccc.edu Fri Feb 2 13:55:35 2007 } 4, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) } 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } l12JtY1x025400 } 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 13:55:34 -0600 } 4, 17 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) } 4, 17 -- (authenticated bits=0) } 4, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id l12JtYEb022147 } 4, 17 -- for {Microscopy-at-Microscopy.Com} ; Fri, 2 Feb 2007 14:55:34 -0500 } (EST) } 4, 17 -- Message-ID: {45C39735.3020002-at-fccc.edu} } 4, 17 -- Date: Fri, 02 Feb 2007 14:55:33 -0500 } 4, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} } 4, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) } 4, 17 -- MIME-Version: 1.0 } 4, 17 -- To: Microscopy-at-Microscopy.Com } 4, 17 -- Subject: SEM of cells in monolayer } 4, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 17 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 23 -- From dsherman-at-purdue.edu Fri Feb 2 16:28:36 2007 9, 23 -- Received: from 1061exfe02.adpc.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12MSa2o019001 9, 23 -- for {microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 16:28:36 -0600 9, 23 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe02.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 9, 23 -- Fri, 2 Feb 2007 17:28:36 -0500 9, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) 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 ; 9, 23 -- Fri, 2 Feb 2007 22:28:36 +0000 9, 23 -- User-Agent: Microsoft-Entourage/11.3.3.061214 9, 23 -- Date: Fri, 02 Feb 2007 17:28:35 -0500 9, 23 -- Subject: Re: [Microscopy] SEM of cells in monolayer 9, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 9, 23 -- To: {Michal.Jarnik-at-fccc.edu} , 9, 23 -- "message to: MSA list" {microscopy-at-microscopy.com} 9, 23 -- Message-ID: {C1E92543.18A02%dsherman-at-purdue.edu} 9, 23 -- Thread-Topic: [Microscopy] SEM of cells in monolayer 9, 23 -- Thread-Index: AcdHGXoBuG18erMMEdu8XQARJN08Mg== 9, 23 -- In-Reply-To: {200702021959.l12JxRoe029306-at-ns.microscopy.com} 9, 23 -- Mime-version: 1.0 9, 23 -- Content-type: text/plain; 9, 23 -- charset="US-ASCII" 9, 23 -- Content-transfer-encoding: 7bit 9, 23 -- X-OriginalArrivalTime: 02 Feb 2007 22:28:36.0656 (UTC) FILETIME=[7AFDDB00:01C74719] ==============================End of - Headers==============================
==============================Original Headers============================== 19, 25 -- From Ann.Lehman-at-trincoll.edu Fri Feb 2 17:47:55 2007 19, 25 -- Received: from wsmtp1.cc.trincoll.edu (wsmtp2.cc.trincoll.edu [157.252.10.109]) 19, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l12NlsLt031670 19, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 17:47:55 -0600 19, 25 -- Received: from hockberry.cc.trincoll.edu ([157.252.15.76]) by wsmtp1.cc.trincoll.edu with Microsoft SMTPSVC(6.0.3790.1830); 19, 25 -- Fri, 2 Feb 2007 18:47:56 -0500 19, 25 -- Received: from exbe1.cmpcntr.tc.trincoll.edu ([157.252.15.111]) by hockberry.cc.trincoll.edu with Microsoft SMTPSVC(6.0.3790.1830); 19, 25 -- Fri, 2 Feb 2007 18:47:56 -0500 19, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 19, 25 -- Content-class: urn:content-classes:message 19, 25 -- MIME-Version: 1.0 19, 25 -- Content-Type: text/plain; 19, 25 -- charset="US-ASCII" 19, 25 -- Subject: RE: [Microscopy] Re: SEM of cells in monolayer 19, 25 -- Date: Fri, 2 Feb 2007 18:47:55 -0500 19, 25 -- Message-ID: {8CF6A92CB628444FB3C757618CD2803929B4DC-at-exbe1.cmpcntr.tc.trincoll.edu} 19, 25 -- X-MS-Has-Attach: 19, 25 -- X-MS-TNEF-Correlator: 19, 25 -- Thread-Topic: [Microscopy] Re: SEM of cells in monolayer 19, 25 -- thread-index: AcdHGehv3GW/kzqFSiGJFyf2s1gJngACcitQ 19, 25 -- From: "Lehman, Ann R" {Ann.Lehman-at-trincoll.edu} 19, 25 -- To: {Microscopy-at-microscopy.com} 19, 25 -- X-OriginalArrivalTime: 02 Feb 2007 23:47:56.0185 (UTC) FILETIME=[8FE45890:01C74724] 19, 25 -- Content-Transfer-Encoding: 8bit 19, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l12NlsLt031670 ==============================End of - Headers==============================
Hi Michal, When I did cultured cells on coverslips found problems like you were experiencing.
1. We changed to Parducz fixative instead of Osmium which is a wonderful hardener for filipodia, cilia, etc. and was used mainly in the LM until SEM preps came along. Parducz is 6 parts of 2% osmium(aq) to 1 part sat'd HgCL2. (HgCl2 can be made by merely dumping in some HgCl2 into distilled water until it doesn't dissolve and go a bit further until there is a small amt of ppt. on the bottom. We kept it in a brown bottle and kept at room temp for many months. Take only from the top of the bottle.) Mix it together just before use. Thus mix for instance 12 ml. of 2% aq Os and 2 ml HgCl2. We only used the mixture once and then properly disposed of it. We often didn't bother with the glut as the Parducz did the trick, but if we had to keep the cells before they were prepared then we kept them in glut. (Something like a 3% glut in a non-phosphate buffer. Phosphates ppt for SEM.) Time: 1-2 hrs in glut (unless stored) and then 1 hr in Parducz then either freeze drying or CPD after proper dehydration in EtOH. We never went through amyl acetate. OR we fixed directly into Parducz for 1 hr then dehydration and CPD or FD. (FD doesn't require dehydration and we went directly from the Parducz - quickly froze it, then Pearse Tissue Dryer)
2. Also check how quickly your CPD is releasing pressure. If it is more than 100 psi per minute then it is likely doing the damage.
3. Agree with the other dehydration comments.
Good Luck,
Judy
Judy Murphy, PhD microscopyproducts Stockton, CA
Michal.Jarnik-at-fccc.edu wrote:
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==============================Original Headers============================== 12, 19 -- From murphyjudy-at-comcast.net Fri Feb 2 22:16:22 2007 12, 19 -- Received: from alnrmhc14.comcast.net (alnrmhc14.comcast.net [204.127.225.94]) 12, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l134GMnb014473 12, 19 -- for {microscopy-at-microscopy.com} ; Fri, 2 Feb 2007 22:16:22 -0600 12, 19 -- Received: from [192.168.1.5] (c-67-181-85-102.hsd1.ca.comcast.net[67.181.85.102]) 12, 19 -- by comcast.net (alnrmhc14) with ESMTP 12, 19 -- id {20070203041621b1400emu06e} ; Sat, 3 Feb 2007 04:16:21 +0000 12, 19 -- Message-ID: {45C40C95.8010807-at-comcast.net} 12, 19 -- Date: Fri, 02 Feb 2007 20:16:21 -0800 12, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 12, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 12, 19 -- X-Accept-Language: en-us, en 12, 19 -- MIME-Version: 1.0 12, 19 -- To: Michal.Jarnik-at-fccc.edu, Microscopy List Server {microscopy-at-microscopy.com} 12, 19 -- Subject: Re: [Microscopy] SEM of cells in monolayer 12, 19 -- References: {200702021959.l12JxY0W029458-at-ns.microscopy.com} 12, 19 -- In-Reply-To: {200702021959.l12JxY0W029458-at-ns.microscopy.com} 12, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nbyers23-at-msn.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, February 3, 2007 at 17:08:16 ---------------------------------------------------------------------------
Email: nbyers23-at-msn.com Name: Nanci Byers
Organization: Weber State University
Education: Undergraduate College
Location: Ogden, Utah USA
Question: I am trying to find some additional materials on learning how to properly draw to scale when viewing slides through a microscope. My professor went over the concept very quickly and I would like some additional information. I am currently taking a botany class.
How refreshing to hear of a botany professor who still teaches looking down a microscope tube and drawing! It seems these days teachers prefer to interface a digital camera to a microscope and broadcast it to students' laptops or something. It takes me back to undergrad school in the early 70s when my botany prof taught us how to stipple. (I'm not just being maudlin; I really think putting something on a slide with your own hands and putting it under the lens and focusing on it just makes it seem real). I guess that when you say draw to scale you mean keeping proportions accurate in two dimensions, in which case you could use an eyepiece reticle grid and make your drawing on a similar grid on your paper. If you're wondering about how to figure out how many times bigger your drawing is than the actual object, you could put a specimen of a known size (say, a finely graduated machinist's ruler or whatever) under the lens, then look at it through the eyepiece while using your other eye to look at another ruler placed on your drawing surface, superimposing the images, then divide the actual known dimension on the microscope stage into the equivalent dimension on your paper to arrive at the number of times your drawing is magnified. I hope this makes sense, or that someone else can explain it more succinctly or tell you a better method. Cheers! And kudos to your teacher!
Paul Grover
--- nbyers23-at-msn.com wrote:
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==============================Original Headers============================== 8, 20 -- From pbgrover-at-yahoo.com Sat Feb 3 20:17:25 2007 8, 20 -- Received: from web34205.mail.mud.yahoo.com (web34205.mail.mud.yahoo.com [66.163.178.120]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l142HPPl005954 8, 20 -- for {microscopy-at-microscopy.com} ; Sat, 3 Feb 2007 20:17:25 -0600 8, 20 -- Received: (qmail 1780 invoked by uid 60001); 4 Feb 2007 02:17:24 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=X-YMail-OSG:Received:Date:From:Reply-To:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 8, 20 -- b=mvL/iFkUCrg8bm8jYFs5tDfJIfp4CGjhsTrisscdG+EYGH1Vl9egoZ2cLDiM7DLbHDTEqzXiI3/cecd+IrB3lm7AQBbSPTs80JRJJ4XBsC0Zqt7TV2FbvQ/PtpC52a2cnCcr33NLmbEdpcg0HuPLcGUn/1j14dOM0/M//R08pSc=; 8, 20 -- X-YMail-OSG: VxGEWtMVM1kiq0XbRru_.RRq7wnfQleMed_eQYoNKATBr5KuqJ4EBUwEU7twKZ2COtlvQtwFC9yNNyy1pm7n7HHPntDMokso7JJrQx2mi2yhoSQdLVLezAwuV_5s3ICLh2uJ19pq9oJ_rB4- 8, 20 -- Received: from [74.140.104.200] by web34205.mail.mud.yahoo.com via HTTP; Sat, 03 Feb 2007 18:17:24 PST 8, 20 -- Date: Sat, 3 Feb 2007 18:17:24 -0800 (PST) 8, 20 -- From: paul grover {pbgrover-at-yahoo.com} 8, 20 -- Reply-To: pbgrover-at-yahoo.com 8, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: how to properly draw to scale 8, 20 -- To: microscopy-at-microscopy.com 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit 8, 20 -- Message-ID: {675137.1361.qm-at-web34205.mail.mud.yahoo.com} ==============================End of - Headers==============================
--| --|Dear Listers, --| --|I am about to prepare some zebra fish heads for SEM - to look at morphology --|& --|take some measurements. What do you think are the pros & cons of critical --|point drying versus HMDS drying after fixation in GDA+PVP in SCB followed by --|osmium+potassium ferrocyanide followed by staining in aqueous UA before --|dehydration for small fish heads? I have had success with the HMDS drying --|using cultured cells and insect tissue. The thermocirculator attached to my --|CPD has problems - it is difficult to heat slowly enough so until I can --|replace it I thought HMDS might be an alternative method. --| --| --|Many thanks --|Ursula --|--------------- --| --|Ursula J. Potter --|Centre for Electron Optical Studies (CEOS) --|Building 3 West 2.15 --|The University of Bath --|Claverton Down --|Bath BA2 7AY --|UK --|Tel: 01225 385651 --|Email: U.J.Potter-at-bath.ac.uk
Dear Ursula,
If you want to make dimensional measurements on dried tissue, I strongly recommend that you take the time to read
Boyde, A, Maconnachie, E. (1979) Volume changes during preparation of mouse embryonic tissue for scanning electron microscopy, Scanning 2:149-163
Boyde, A, Maconnachie, E, (1981) Morphological correlations with dimensional change during SEM specimen preparation, Scanning Electron Microsc. 1981, IV:27-34
These are the only two publications of which I am aware where the authors actually made dimensional measurements of soft tissue (embryos) as they proceeded through all the stages of fixation, dehydration, and CPD or other drying.
The BEST they got was only 60% volume shrinkage (i.e., the final volume was 40% of the live volume. This works out to be about 25% linear shrinkage.).
This dirty secret often goes unremarked because, as tissue-culture cells are tacked down to glass slides, the x-y dimensional are stabilized by the glass. The thickness shrinks but that is harder to measure.
Of course, I am sure that some tissues may be more robust but I can also assure you that many are much more sensitive. And as I noted, 60% was the BEST they got. There were many ways to make it worse.
Freeze drying was better, especially if you kept the tissue frozen to about -100deg C while you looked at it. but this has its own problems, particularly those to do with ice crystals.
As many readers will likely find my claims "ridiculous," I do encourage you to read the papers and then do you own tests.
Good luck!
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 16-28, 2007, UBC, Vancouver Canada Info: http:// www.3dcourse.ubc.ca/ Applications due by March 15, 2007 "If it ain't diffraction, it must be statistics." Anon.
==============================Original Headers============================== 17, 26 -- From jbpawley-at-wisc.edu Sun Feb 4 14:45:23 2007 17, 26 -- Received: from smtpauth.wiscmail.wisc.edu (adsum.doit.wisc.edu [144.92.197.210]) 17, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l14KjNIO016510 17, 26 -- for {Microscopy-at-Microscopy.Com} ; Sun, 4 Feb 2007 14:45:23 -0600 17, 26 -- Received: from avs-daemon.smtpauth1.wiscmail.wisc.edu by 17, 26 -- smtpauth1.wiscmail.wisc.edu 17, 26 -- (Sun Java System Messaging Server 6.2-7.05 (built Sep 5 2006)) 17, 26 -- id {0JCY00I01HNMZQ00-at-smtpauth1.wiscmail.wisc.edu} for 17, 26 -- Microscopy-at-Microscopy.Com; Sun, 04 Feb 2007 14:45:22 -0600 (CST) 17, 26 -- Received: from [172.16.1.43] ([76.210.70.232]) by smtpauth1.wiscmail.wisc.edu 17, 26 -- (Sun Java System Messaging Server 6.2-7.05 (built Sep 5 2006)) 17, 26 -- with ESMTPSA id {0JCY00HKAHNKC600-at-smtpauth1.wiscmail.wisc.edu} for 17, 26 -- Microscopy-at-Microscopy.Com; Sun, 04 Feb 2007 14:45:21 -0600 (CST) 17, 26 -- Date: Sun, 04 Feb 2007 14:45:18 -0600 17, 26 -- From: James Pawley {jbpawley-at-wisc.edu} 17, 26 -- Subject: Re: SEM of Zebra fish 17, 26 -- X-Sender: jbpawley-at-wiscmail.wisc.edu 17, 26 -- To: "ListServer-at-MSA.Microscopy.Com" {Microscopy-at-Microscopy.Com} 17, 26 -- Message-id: {p0611041bc1ebf5c7eae4-at-[172.16.1.43]} 17, 26 -- MIME-version: 1.0 17, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 17, 26 -- Content-transfer-encoding: 7BIT 17, 26 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=76.210.70.232 17, 26 -- X-Spam-PmxInfo: Server=avs-1, Version=5.2.1.279297, 17, 26 -- Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.2.4.123434, 17, 26 -- SenderIP=76.210.70.232 ==============================End of - Headers==============================
Let me chime in here and completely agree with Jim. I have measured shrinkage on the order of 50% to 60% in soft tissues, in this case neuromast end organs from fish lateral lines. The original sizes were measured both by simple measuring of the intact end organs in a light microscope, and measuring the impressions the end organs made in silicone casts of the lateral lines. These two measurements were in almost exact agreement. The end organs as seen in the SEM were at best 1/2 the size of the living and the "fixed-only", i.e., not dehydrated, organs. Further the shrinkage was *not* isometric. That is, the shrinkages along X and Y axis imposed onto the end organs were different. Keep in mind that tissues have mechanical properties, and these properties depend on the histology of the tissue. E.g., what kind of collagen does the tissue have? In what directions do the fibers run? Shrinkage in the direction of the fibers will be different than shrinkage in the direction orthogonal to the fiber direction. And that's just the begining. I don't know of any studies on such tissue properties and how they affect dimensional changes. If there are such references, I would very much appreciate it if they were posted to the list. Best answer to measuring zebra heads: do all your measurements with a light microscope. Make yourself a little jig, so the heads can all be held in exactly the same set of positions so all the measurements will be correct. Otherwise you'll have measurement errors caused by slightly different tilts of the heads. Do this before fixation. A rule of thumb used by fish ecologists, back when I was doing marine ecology, was that all formalin-fixed fish lengths were 10% too short. That might not be really true, but the shrinkage was.
Phil
} --| } --|Dear Listers, } --| } --|I am about to prepare some zebra fish heads for SEM - to look at morphology } --|& } --|take some measurements. What do you think are the pros & cons of critical } --|point drying versus HMDS drying after fixation in GDA+PVP in SCB } followed by } --|osmium+potassium ferrocyanide followed by staining in aqueous UA before } --|dehydration for small fish heads? I have had success with the HMDS drying } --|using cultured cells and insect tissue. The thermocirculator attached to my } --|CPD has problems - it is difficult to heat slowly enough so until I can } --|replace it I thought HMDS might be an alternative method. } --| } --| } --|Many thanks } --|Ursula } --|--------------- } --| } --|Ursula J. Potter } --|Centre for Electron Optical Studies (CEOS) } --|Building 3 West 2.15 } --|The University of Bath } --|Claverton Down } --|Bath BA2 7AY } --|UK } --|Tel: 01225 385651 } --|Email: U.J.Potter-at-bath.ac.uk } } } Dear Ursula, } } If you want to make dimensional measurements on dried tissue, I } strongly recommend that you take the time to read } } Boyde, A, Maconnachie, E. (1979) Volume changes during preparation of } mouse embryonic tissue for scanning electron microscopy, Scanning } 2:149-163 } } Boyde, A, Maconnachie, E, (1981) Morphological correlations with } dimensional change during SEM specimen preparation, Scanning Electron } Microsc. 1981, IV:27-34 } } } These are the only two publications of which I am aware where the } authors actually made dimensional measurements of soft tissue } (embryos) as they proceeded through all the stages of fixation, } dehydration, and CPD or other drying. } } The BEST they got was only 60% volume shrinkage (i.e., the final } volume was 40% of the live volume. This works out to be about 25% } linear shrinkage.). } } This dirty secret often goes unremarked because, as tissue-culture } cells are tacked down to glass slides, the x-y dimensional are } stabilized by the glass. The thickness shrinks but that is harder to } measure. } } Of course, I am sure that some tissues may be more robust but I can } also assure you that many are much more sensitive. And as I noted, } 60% was the BEST they got. There were many ways to make it worse. } } Freeze drying was better, especially if you kept the tissue frozen to } about -100deg C while you looked at it. but this has its own } problems, particularly those to do with ice crystals. } } As many readers will likely find my claims "ridiculous," I do } encourage you to read the papers and then do you own tests. } } Good luck! } } 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 16-28, 2007, UBC, Vancouver Canada } Info: http:// www.3dcourse.ubc.ca/ Applications due by March 15, 2007 } "If it ain't diffraction, it must be statistics." Anon. -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Mon Feb 5 07:32:34 2007 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l15DWYqm013008 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 5 Feb 2007 07:32:34 -0600 4, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l15DxQnU008863 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 5 Feb 2007 08:59:32 -0500 4, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 22 -- Mon, 5 Feb 2007 08:32:28 -0500 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230903c1ecdf4983fa-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200702042050.l14Kolif023703-at-ns.microscopy.com} 4, 22 -- References: {200702042050.l14Kolif023703-at-ns.microscopy.com} 4, 22 -- Date: Mon, 5 Feb 2007 08:32:27 -0500 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: [Microscopy] Re: SEM of Zebra fish 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 05 Feb 2007 13:32:28.0736 (UTC) FILETIME=[14A80C00:01C7492A] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -3.1 () FCS_URI_NODOTS,J_CHICKENPOX_33,L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Michal, I would differ only slightly with Phil's suggestions: since your cells are on or in collagen, you will probably need to extend your dehydrations: perhaps 2 changes at each concentration, starting at 50% or even 30% as Phil suggested. Collagen is an incredible sponge and I've had a miserable time with it over the years. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
I agree, any diminishing of visual acuity is indeed a disturbing event for microscopists; however, I don't think age is necessarily a primary factor involved in causing eye floaters. I am nearly 84 years old, and don't have any floaters (however I seem to recall having a few sometime in the distant past). The only thing I can think of that might be contributing to this is the fact that I take large amounts of vitamins of all types, about half the amounts recommended by Linus Pauling in his book How to Live Longer and Feel Better, and have been doing so for more than 20 years.
My problem now seems to be the gradual development of cataracts, which my doctor tells me is a process definitely related to aging. It may be that the vitamins (especially vitamin C) are slowing this precess down, but I'm afraid I'll eventually need to have the cataracts removed. -- 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============================== 2, 14 -- From bigelow-at-engin.umich.edu Mon Feb 5 15:19:12 2007 2, 14 -- Received: from srvr20.engin.umich.edu (srvr20.engin.umich.edu [141.213.75.22]) 2, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l15LJCCo016487 2, 14 -- for {microscopy-at-microscopy.com} ; Mon, 5 Feb 2007 15:19:12 -0600 2, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 2, 14 -- by srvr20.engin.umich.edu (8.13.6/8.13.6) with ESMTP id l15LJB6k009826 2, 14 -- for {microscopy-at-microscopy.com} ; Mon, 5 Feb 2007 16:19:11 -0500 (EST) 2, 14 -- Mime-Version: 1.0 2, 14 -- Message-Id: {p06210202c1ed4c72bb2b-at-[141.212.131.221]} 2, 14 -- Date: Mon, 5 Feb 2007 16:19:10 -0500 2, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 2, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 2, 14 -- Subject: [Microscopy]RE: Eye Floaters 2, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Only 11 days left to submit an abstract for Seeing at the Nanoscale, the fifth annual scientific conference focusing on nanostructural imaging, characterization, and modification using scanning probe microscopy (SPM) and related techniques.
The conference location is Santa Barbara, California, June 24-27, 2007. Sponsored by Veeco Instruments and the California NanoSystems Institute (CNSI) at the University of California, Santa Barbara (UCSB), this two-and-one-half day event includes technical presentations, a nanotechnology poster contest, and a beach barbecue-Santa Barbara style.
Highlighted by Keynote speakers Angela Belcher and David Awschalom, Seeing at the Nanoscale provides an optimum forum for "scientists to speak to scientists" on a wide variety of nanotechnology topics with technical sessions on:
Extending the Limits of SPM
Using AFM and Combined AFM-Optical Techniques to Probe Biological Structures and Forces
Next Generation Materials and Polymer Systems
Beyond Topography: Measurement of Physical Properties at the Nanoscale - Nanomechanical, Local Property, Electrical, Optical, Magnetic & Thermal
Instruments and Probes - New Tools & Techniques for Nanoscience
To submit your abstract, review submission guidelines, and learn more about the conference, visit www.veeco.com/nanoconference.
Take part as a presenter in the industry's most dynamic conference!
Veeco Instruments & CNSI
_________________________________ Marlene Carlyle Conference Coordinator Veeco Instruments 112 Robin Hill Road Santa Barbara, CA 93117 Phone: 805-967-1400 Fax: 805-967-7717 mcarlyle-at-veeco.com _________________________________
==============================Original Headers============================== 14, 20 -- From MCarlyle-at-veeco.com Mon Feb 5 15:34:37 2007 14, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.VEECO.com [68.111.35.167]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l15LYafD027814 14, 20 -- for {Microscopy-at-Microscopy.com} ; Mon, 5 Feb 2007 15:34:37 -0600 14, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 14, 20 -- content-class: urn:content-classes:message 14, 20 -- MIME-Version: 1.0 14, 20 -- Content-Type: text/plain; 14, 20 -- charset="US-ASCII" 14, 20 -- Subject: SPM - Abstract Submission Deadline for Seeing at the Nanoscale Conference 14, 20 -- Date: Mon, 5 Feb 2007 13:34:36 -0800 14, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F42012002EB-at-sboexch2.int.veeco.com} 14, 20 -- X-MS-Has-Attach: 14, 20 -- X-MS-TNEF-Correlator: 14, 20 -- Thread-Topic: SPM - Abstract Submission Deadline for Seeing at the Nanoscale Conference 14, 20 -- Thread-Index: AcdJbW/FwOsKhybpQgW4cozElHmuCg== 14, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 14, 20 -- To: {Microscopy-at-Microscopy.com} 14, 20 -- Content-Transfer-Encoding: 8bit 14, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l15LYafD027814 ==============================End of - Headers==============================
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Email: mike_microscopy-at-yahoo.com Name: Mike
Title-Subject: [Filtered] Looking for simulation software for CTEM image
Question: I am looking for Windows-based software that allows us to simulate bright-field and dark-field images. The images to be simulated are about the fringes observed at the interface with known elastic strains. All atomic positions, including the atoms located on both sides of the interface and the atoms suffering from the elastic stains at the interface, and the indices of the interface are available. The orientation of the crystals is also known. In this way, the new unit cell involving the interface and the atoms on both sides of the interface can be built. I am a material researcher, not familiar with the principles used in software. So hoping the software is of user friendly interface. It is preferential that just inputting all atomic coordinates relative to the new unit cell and giving the observed orientation and the thickness of the specimen the bright-field and dark-field images can be simulated and output. Please let me know where to get such simulation software. Any input would be highly appreciated!
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Email: camiller-at-anatomy.iupui.edu Name: Caroline Miller
Organization: Indiana Microscopy Society
Title-Subject: [Filtered] Joint LAS Spring Meeting
Question: There will be a joint LAS Spring Meeting, "Imaging for Nanotechnology," hosted by the Indiana Microscopy Society in Indianapolis, IN (home of the 2006 NFL Super Bowl Champions!)on April 20th and 21st, 2007. Co-sponsored by Central States, Iowa, Midwest and Michigan LAS. Friday's session, April 20th will include five speakers, Dr. M. Marko, Dr. W. Landis, R. Gursky, Dr. J. Pawley and Dr. D. Newbury with an evening social at the "White River Gardens." Saturday's session, April 21st will be a half day of workshops, demos and tours of the Electron Microscoppy Center and the Indiana Center for Biological Microscopy. Awards will be given for the best poster either by a student or faculty/staff and for the best Biological and Physical Science Micrograph. There is a discount if you pre-register before March 15th. Abstract deadline is March 1st. Check out the INMS web site for a complete program, registration form, accomodations and application for a student fellowship. www.indianamicroscopy.org Send your abstract or questions to the Program Chair, Caroline Miller, camiller-at-anatomy.iupui.edu, fax # 317-278-2040.
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Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers Institute
Title-Subject: [Filtered] Tousimis Samdri
Question: Good Afternoon,
I am looking for a distributor of the Tousimis Samdri Critical Point Dryer for the Kansas City, Missouri area.
A colleague of mine was looking to lower his costs on maintenance of his Jeol SEM. He searched for an EM maintenance course and found the company Protrain. We scheduled a 5 day training course with Stephen Chapman of Protrain. and that was the best invest we ever made. Months before the training was to begin we were asked what scopes we had and Steve then tailored a course that he taught us at our site on our scopes. Steve taught us normal scope maintenance and he taught us how to trouble shoot not just the column but user induced problems. Mr. Chapman scripted customized operating instructions for the TEM in our facilities as we operated the microscope in addition to a digital book on Electron Microscopy. We were also showed how to improve the safety of our laboratories and core facility.
Don Johnson Physics Department University of Maryland Baltimore County
Chere Petty Department of Biological Sciences University of Maryland Baltimore County
We have no financial interest in Protain.
-- Chere Petty, M.S. Manager of Keith R. Porter Imaging Facility Department of Biological Sciences University of Maryland Baltimore County (UMBC) 1000 Hilltop Circle Baltimore, Maryland 21250 Fax: 410-455-3875 Cell: 301-367-8408 cpetty1-at-umbc.edu www.emumbc.com
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Dear Materials Characterization Specialist: Here is an open job at University of California. Please apply it by following the instruction below. Thanks for your attention. Jian-Guo
Materials Characterization Specialist University of California, Irvine Salary: Commensurate with experience
The University of California, Irvine is seeking a materials characterization specialist to work in the campus-wide Nanomaterials Characterization and Fabrication Facility (NCF2). The successful candidate is expected to have an earned PhD degree in a relevant field, possess an extensive knowledge in analytical instrumentation (SEM, XRD, AFM, TEM, FTIR, TGA, DSC) and research implementation, and have rich experience in sample preparation. He/she should have either extensive knowledge of techniques and protocols in soft materials characterization, or demonstrate a desire to acquire such knowledge. The applicant should also have excellent writing and inter-personal communication skills, and strong team spirit. Good computing skills are also desirable. The materials characterization specialist's responsibilities include carrying out day-to-day operations of analytical equipments including, but not limited to, SEM, XRD, TEM and AFM/STM. He/she will also be responsible for (1) maintaining all instruments in good working condition, (2) training and helping users in the use of analytical equipments, (3) preparing samples and providing help to users in sample preparation, (4) assisting in courses related to analytical instrumentation, (5) conducting service for off-campus and industrial users, (6) maintaining the facility infrastructure, and (7) carrying out miscellaneous facility-related tasks assigned by the facility director. The specialist will report to the director of NCF2. Salary is commensurate with experience. This position will open immediately and remain open until filled.
Please include in the application the following materials: • Curriculum Vita • 2-3 publications • Three letters of recommendation letters (may be submitted shortly after the submission of the application)
Ms. Dorothy Miles 4100 Calit2 Bldg Irvine, CA 92697-2800 djmiles-at-uci.edu ph: 949/824-5178 fax: 949/824-4403
The University of California, Irvine is an equal opportunity employer committed to excellence through diversity.
==============================Original Headers============================== 10, 20 -- From jzheng-at-uci.edu Tue Feb 6 22:04:02 2007 10, 20 -- Received: from relay2.es.uci.edu (relay2.es.uci.edu [128.200.80.28]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17441PS001387 10, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Feb 2007 22:04:02 -0600 10, 20 -- Received: from webmail.uci.edu (webmail1.es.uci.edu [128.200.80.36]) 10, 20 -- by relay2.es.uci.edu (8.13.1/8.13.1) with ESMTP id l1743x9X006935 10, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Feb 2007 20:04:01 -0800 10, 20 -- Received: from 128.195.177.193 10, 20 -- (SquirrelMail authenticated user jzheng) 10, 20 -- by webmail.uci.edu with HTTP; 10, 20 -- Tue, 6 Feb 2007 20:04:01 -0800 (PST) 10, 20 -- Message-ID: {2970.128.195.177.193.1170821041.squirrel-at-webmail.uci.edu} 10, 20 -- Date: Tue, 6 Feb 2007 20:04:01 -0800 (PST) 10, 20 -- Subject: Good opportunity for Materials Characterization Specialist 10, 20 -- From: jzheng-at-uci.edu 10, 20 -- To: Microscopy-at-microscopy.com 10, 20 -- User-Agent: SquirrelMail/1.4.9a 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain;charset=iso-8859-1 10, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (miranda.s.norby-at-sendit.nodak.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, February 6, 2007 at 14:24:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both miranda.s.norby-at-sendit.nodak.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: miranda.s.norby-at-sendit.nodak.edu Name: Miranda Norby
Organization: Rolette Public School
Education: Graduate College
Location: Rolette, ND
Question: I teach 7-12 science-classes include Earth Science, Biology, Chemistry, Physics. We are in the process of ordering student microscopes. I was hoping to receice some recommendations as to products that you have found to be student friendly and durable. Thank you for any imput you can provide.
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Email: acorn1800-at-yahoo.com Name: Wayne
Title-Subject: [Filtered] Nikon EFM microscope camera
Question: I have recently aquired a Nikon Microscope Camera, model EFM. I cannot find the battery compartment. I would also like a copy of the operating manual, I can pay for copying and shipping costs.
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Email: dlowry-at-asu.edu Name: David Lowry
Organization: Arizona State University
Title-Subject: [Filtered] critical point drying question
Question: Is ethanol miscible in liquid CO2? I can't seem to find anywhere this is explicitly stated. I have seen SEM protocols which imply they use EtOH as dehydrating agent and then directly CPD with CO2. We have an older Balzers CPD020 unit and the manual has a flow-chart which shows only acetone or amyl acetate as dehydrants prior to CO2 transition. It is my understanding that EtOH is a weaker organic solvent than acetone, and if EtOH mixes with CO2, this may be useful for certain CPD applications. Thanks
We have used ethanol followed by liquid carbon dioxide for years in our Tousimis Samdri-780 CPD. I switched from acetone, which I have been told mixes better than ethanol with liquid carbon dioxide, to reduce extraction from samples.
Dave
-----Original Message----- X-from: dlowry-at-asu.edu [mailto:dlowry-at-asu.edu] Sent: 07 February 2007 07:18 To: David Patton
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both dlowry-at-asu.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: dlowry-at-asu.edu Name: David Lowry
Organization: Arizona State University
Title-Subject: [Filtered] critical point drying question
Question: Is ethanol miscible in liquid CO2? I can't seem to find anywhere this is explicitly stated. I have seen SEM protocols which imply they use EtOH as dehydrating agent and then directly CPD with CO2. We have an older Balzers CPD020 unit and the manual has a flow-chart which shows only acetone or amyl acetate as dehydrants prior to CO2 transition. It is my understanding that EtOH is a weaker organic solvent than acetone, and if EtOH mixes with CO2, this may be useful for certain CPD applications. Thanks
I routinely use acetone in a Balzers critical point drier but sometimes want to dry samples that may be damaged by the acetone. In these cases (for example preserving the outer surfaces of insect eggs prior to SEM examination) I use ethanol. From my experience, the ethanol is nowhere near as miscible as acetone in liquid CO2 but still works perfectly well.
You need to use more exchanges of the liquid CO2 in order to replace the ethanol, leaving to stand for a few minutes between each exchange. If your CPD system has a stirrer, even better! I find that the drying process in the CPD with ethanol can take around four times longer than the conventional technique using acetone. However as long as it achieves the result, then the additional time has been worthwhile.
Best of luck!
Regards,
Chris
Chris Jones ----------------------------------- Hitachi High-Technologies 7 Ivanhoe Road Hogwood Industrial Estate Finchampstead Wokingham Berks RG40 4QQ Tel +44(0)118 932 8632 Fax +44(0)118 973 2622 chrisj-at-hitachi-hitec-uk.com www.hitachi-hitec-uk.com
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This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both dlowry-at-asu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dlowry-at-asu.edu Name: David Lowry
Organization: Arizona State University
Title-Subject: [Filtered] critical point drying question
Question: Is ethanol miscible in liquid CO2? I can't seem to find anywhere this is explicitly stated. I have seen SEM protocols which imply they use EtOH as dehydrating agent and then directly CPD with CO2. We have an older Balzers CPD020 unit and the manual has a flow-chart which shows only acetone or amyl acetate as dehydrants prior to CO2 transition. It is my understanding that EtOH is a weaker organic solvent than acetone, and if EtOH mixes with CO2, this may be useful for certain CPD applications. Thanks
==============================Original Headers============================== 6, 12 -- From zaluzec-at-microscopy.com Wed Feb 7 01:13:47 2007 6, 12 -- Received: from [10.2.6.132] (msdvpn8.msd.anl.gov [130.202.238.72]) 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l177Djd1019271 6, 12 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 01:13:46 -0600 6, 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- Message-Id: {p06020402c1ef2e29797d-at-[10.2.6.132]} 6, 12 -- Date: Wed, 7 Feb 2007 20:20:36 +1300 6, 12 -- To: microscopy-at-microscopy.com 6, 12 -- From: dlowry-at-asu.edu (by way of MicroscopyListserver) 6, 12 -- Subject: viaWWW: critical point drying question 6, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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==============================Original Headers============================== 33, 21 -- From chrisj-at-hitachi-hitec-uk.com Wed Feb 7 03:49:40 2007 33, 21 -- Received: from MS-RD-1.hitachi-hitec-uk.com ([195.92.107.19]) 33, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l179ndkO030295 33, 21 -- for {Microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 03:49:40 -0600 33, 21 -- Received: from mailserver2.hitachi-hitec-uk.com (ld-rd-1 [172.21.136.100]) by 33, 21 -- MS-RD-1.hitachi-hitec-uk.com (Clearswift SMTPRS 5.1.4) with ESMTP id 33, 21 -- {T7da8046cd8ac1588603c0-at-MS-RD-1.hitachi-hitec-uk.com} ; Wed, 7 Feb 33, 21 -- 2007 09:49:34 +0000 33, 21 -- Subject: Re: [Microscopy] viaWWW: critical point drying question 33, 21 -- To: dlowry-at-asu.edu 33, 21 -- Cc: Microscopy-at-microscopy.com 33, 21 -- X-Mailer: Lotus Notes Release 5.0.8 June 18, 2001 33, 21 -- Message-ID: {OF261C423E.9E13CC21-ON8025727B.00346DAD-at-hitachi-hitec-uk.com} 33, 21 -- From: chrisj-at-hitachi-hitec-uk.com 33, 21 -- Date: Wed, 7 Feb 2007 09:49:22 +0000 33, 21 -- X-MIMETrack: Serialize by Router on 33, 21 -- LD-RD-1/RD/HHT-UK(Release 5.0.12 |February 13, 2003) at 07/02/2007 33, 21 -- 09:49:23 33, 21 -- MIME-Version: 1.0 33, 21 -- Content-type: text/plain; charset="us-ascii" 33, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
We get asked this fairly regularly by schools and parents (so we have stock replies). Often professional microscopists aren't the best source of information for cheap microscope's as all our optical systems are always over £10,000, and most lab mainstays like our three confocal microscopes come in at nearer £250,000 each (and users still complain about image quality).
Cheap microscopes under £100 are always a disappointment and toy (often called student) microscopes can be very poor. For home use or a demonstration model, you can easily buy second-hand via ebay, but again there are risks that the optics will be damaged or simply very dirty and difficult to clean and you may make an expensive mistake. Look for old branded 'laboratory' microscopes e.g. Bausch & Lomb, Prior, Leitz, Reichert, Baker, but not Tasco toys. Famous existing brands like Zeiss and Olympus attract a high premium. The sellers are often not microscopists though, and many are sold as collector's items and not intended for 'scientific' use. However many home users often get bored with new microscopes after a while, so there is a second hand market for cheaper school/college lab grade models. Also I would try any local microscope enthusiast clubs - they aren't as common as the many excellent astronomy [telescope] clubs but they are about and have knowledgeable enthusiasts with an eye for low cost quality systems. To purchase for classes you will have to buy new, probably from a schools supplier.
There are suppliers geared up to providing cheaper microscopes for schools & colleges, so you can ask around at other school/college's science departments, but expect to pay nearer £500 each for a quality setup (although with those like bottom end Meade [www.meade.com] at around £100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section - try some on approval?.
For pond life etc. a stereoscopic 'dissecting' microscope (40x to 120x mag) is ideal for home/school use. Also remember of course that you can get a really long way with a good large magnifying glass (not the really small hi-mag cheap folding lens ones, try before you buy) - I have a few excellent ones at home for £1 and a good low mag Osram one that includes an illuminating halogen bulb at £8. In general I would say a well made stereo dissecting microscope is a good choice (if not the best) for younger kids as it's great for viewing living things and enlarges what you can see already - look for 40x rather than 4x though. Decent ones are a bit expensive (£500+). These are ideal for small animals and things like colonies but are totally unsuitable for viewing single cells or sections where something like 100x to 600x is preferred (expensive 20, 40 to 63x objectives), and requires a standard compound microscope.
Generally prepared slides can rapidly get very boring for all ages, but living or unusual things (even hamster fur) always attract an audience. Also try your flatbed film scanner, (not the LiDe types that have a very limited depth of focus, and any from around £60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class of children can look at a computer screen with some pushing and shoving. Best to try them on 'approval' as many cheap microscopes can be very disappointing if you expect too much. For things like living cells/microscopic organisms you would probably want some form on contrast enhancement like Phase Contrast optics that adds to the complexity and expense (and starts to put the microscope towards the £1,000 category). Viewing mostly fixed & stained prepared sections obviously reduces the need for any optical contrast enhancement.
Excellent pre-prepared stained slides of simple organisms, plant stems and leaves or bits of rats, insects etc.. can be bought via ebay, but they tend to be expensive and are easily broken by any age-group. Mounted slides keep well though, so 'vintage' ones even from 50 years ago can still look OK. Again school suppliers are geared up to provide for this sort of thing far more cheaply (and most schools have an aging collection anyway).
At home and our Primary school (under 11) we use the Digital Blue QX-5 (£70)- it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. I have one at home for my kids (boy 10 and girl 12) but it only gets occasional use now the novelty has worn off. See http://micro.magnet.fsu.edu/optics/intelplay/index.html (not updated for the QX-5 but all applies - the site even discusses ways of contrast enhancement etc..). Once on the PC the 640x480 images can be manipulated and pasted etc, and the QX-5 does time-lapse for things like crystal growth. Living plants growing and small animals. This toy has nowhere near the resolution you require though. The similar but far better built Olympus MIC-D was great but being over £500 it was just too expensive for most schools and is now discontinued - there are other similar budget video microscope systems about though.
The macro on a good digital camera (like the image stabilised 5MP Canon S2IS - http://www.dpreview.com/reviews/canons2is)is also worth a try, particularly with a small tripod and halogen bendy desk lamp, if very close-up. I have an E500 digital SLR system with enlargement rings but they are more difficult to use. You can get quite reasonable pictures by resting a small compact digital camera lens against the eyepiece of a microscope. Plus you can use the camera for normal photography when you get bored with microscopy. I wouldn't fancy a large class using my E500 SLR though.
By the way, do try growing crystals on a slide, a few drops of a saturated solution of salt (NaCl) or copper sulphate will grow superb crystals on the surface of a slide when viewed under a microscope (but it takes a few hours for the crystals to form and they often look best before the liquids all gone). Just make sure you don't drive the objective tips into the solution. It's not biology but its fun (see http://micro.magnet.fsu.edu/.
Sorry I can't offer more specific help as our microscopes are rather more expensive than the one you wish to purchase.
Regards
Keith
Try looking in Amazon.com for decent microscope books with lots of pictures - and they have a good customer review system for books and even microscopes (often viewing micrographs in books or on the VDU screen is better than trying to see it yourself down a microscope). Plus try web searches for general sites like these (and for more specific topics) :
http://www.101science.com/Microscope.htm http://micro.magnet.fsu.edu/ http://www.microscopy-uk.org.uk/index.html http://www.btinternet.com/~stephen.durr/ http://www.mccroneatlas.com http://www.diatoms.co.uk [for fun images made of diatoms]
Examples of sources for cheap school grade microscopes in the UK are:
Have an internet search for school/lab suppliers in the US or just ask other school science departments.
Our lab technicians dealt with all the ordering of prepared slides, we just had to make a general request for the item. You can also buy on ebay but that is probably more for collectors of science equipment and quality may vary. However in the UK there are a lot of specialised suppliers of scientific equipment for secondary schools, who supply prepared slides quite cheaply. A small selection is:
-----Original Message----- X-from: miranda.s.norby-at-sendit.nodak.edu [mailto:miranda.s.norby-at-sendit.nodak.edu] Sent: 07 February 2007 07:18 To: keith.morris-at-ucl.ac.uk
This Question was submitted to Ask-A-Microscopist by (miranda.s.norby-at-sendit.nodak.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, February 6, 2007 at 14:24:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both miranda.s.norby-at-sendit.nodak.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: miranda.s.norby-at-sendit.nodak.edu Name: Miranda Norby
Organization: Rolette Public School
Education: Graduate College
Location: Rolette, ND
Question: I teach 7-12 science-classes include Earth Science, Biology, Chemistry, Physics. We are in the process of ordering student microscopes. I was hoping to receice some recommendations as to products that you have found to be student friendly and durable. Thank you for any imput you can provide.
Miranda, If she hasn't already replied to you, contact Caroline Schooly at schooley-at-mcn.org She is a wealth of information at the public school level.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: miranda.s.norby-at-sendit.nodak.edu [mailto:miranda.s.norby-at-sendit.nodak.edu] Sent: Wednesday, February 07, 2007 2:16 AM To: kenconverse-at-qualityimages.biz
This Question was submitted to Ask-A-Microscopist by (miranda.s.norby-at-sendit.nodak.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, February 6, 2007 at 14:24:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both miranda.s.norby-at-sendit.nodak.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: miranda.s.norby-at-sendit.nodak.edu Name: Miranda Norby
Organization: Rolette Public School
Education: Graduate College
Location: Rolette, ND
Question: I teach 7-12 science-classes include Earth Science, Biology, Chemistry, Physics. We are in the process of ordering student microscopes. I was hoping to receice some recommendations as to products that you have found to be student friendly and durable. Thank you for any imput you can provide.
==============================Original Headers============================== 7, 12 -- From zaluzec-at-microscopy.com Wed Feb 7 01:11:45 2007 7, 12 -- Received: from [10.2.6.132] (msdvpn8.msd.anl.gov [130.202.238.72]) 7, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l177BhUX015500 7, 12 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 01:11:44 -0600 7, 12 -- Mime-Version: 1.0 7, 12 -- X-Sender: (Unverified) 7, 12 -- Message-Id: {p06020400c1ef2db05d32-at-[10.2.6.132]} 7, 12 -- Date: Wed, 7 Feb 2007 20:18:34 +1300 7, 12 -- To: microscopy-at-microscopy.com 7, 12 -- From: miranda.s.norby-at-sendit.nodak.edu (by way of Ask-A-Microscopist) 7, 12 -- Subject: AskAMicroscopist: student microscopes 7, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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==============================Original Headers============================== 22, 30 -- From kenconverse-at-qualityimages.biz Wed Feb 7 07:41:19 2007 22, 30 -- Received: from dpmailmta02.doteasy.com (dpmailmta02.doteasy.com [65.61.209.81]) 22, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17DfI26024283 22, 30 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Feb 2007 07:41:19 -0600 22, 30 -- Received: from qualityimages.biz (unverified [192.168.101.16]) 22, 30 -- by dpmailmta02.doteasy.com (DEO) with ESMTP id 56842295-1814644 22, 30 -- for multiple; Wed, 07 Feb 2007 06:21:32 -0800 22, 30 -- Received: from Ken [66.174.79.240] by qualityimages.biz with ESMTP 22, 30 -- (SMTPD32-8.05) id A6EBCD560152; Wed, 07 Feb 2007 05:40:59 -0800 22, 30 -- Received: from unknown [75.196.126.141] by 66.174.79.240; 07 Feb 2007 07:57:38 -0500 22, 30 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 22, 30 -- To: {miranda.s.norby-at-sendit.nodak.edu} 22, 30 -- Cc: "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 22, 30 -- Subject: RE: [Microscopy] AskAMicroscopist: student microscopes 22, 30 -- Date: Wed, 7 Feb 2007 08:40:43 -0500 22, 30 -- Message-ID: {004801c74abd$9d9e7370$8d7ec44b-at-Ken} 22, 30 -- MIME-Version: 1.0 22, 30 -- Content-Type: text/plain; 22, 30 -- charset="us-ascii" 22, 30 -- X-Priority: 3 (Normal) 22, 30 -- X-MSMail-Priority: Normal 22, 30 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 22, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 22, 30 -- In-Reply-To: {200702070716.l177GDn5029041-at-ns.microscopy.com} 22, 30 -- Importance: Normal 22, 30 -- X-IMSTrailer: __IMail_7__ 22, 30 -- X-IP-stats: Incoming Last 0, First 132, in=3160605, out=0, spam=0 ip=192.168.101.16 22, 30 -- X-External-IP: 192.168.101.16 22, 30 -- Content-Transfer-Encoding: 8bit 22, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l17DfI26024283 ==============================End of - Headers==============================
We are advertising for a cryo TEM operator for a new 300KV cryo TEM to be delivered by early April, 2007. Please see our ad on the MSA web site at: http://www.microscopy.org/MSAUnits/PlacementOffice/JobListings.html
or go to the University of MN job site link for more details: http://employment.umn.edu/applicants/Central?quickFind=58119
or you can contact me directly for more information. Regards, Mike ****************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 18 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 MiNTeC node of the National Nanotechnology Infrastructure Network
University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://www.charfac.umn.edu ********************************************************************
==============================Original Headers============================== 5, 22 -- From mboucher-at-umn.edu Wed Feb 7 07:49:55 2007 5, 22 -- Received: from mta-a2.tc.umn.edu (mta-a2.tc.umn.edu [134.84.119.206]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17DntHA002999 5, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 07:49:55 -0600 5, 22 -- Received: from mike (Mike.charfac.umn.edu [160.94.16.142]) 5, 22 -- by mta-a2.tc.umn.edu (UMN smtpd) with ESMTP 5, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 7 Feb 2007 07:49:55 -0600 (CST) 5, 22 -- X-Umn-Remote-Mta: [N] Mike.charfac.umn.edu [160.94.16.142] #+LO+TS+AU+HN 5, 22 -- Reply-To: {mboucher-at-umn.edu} 5, 22 -- From: "Michael L. Boucher" {mboucher-at-umn.edu} 5, 22 -- To: {Microscopy-at-microscopy.com} 5, 22 -- Subject: Cryo TEM operator needed 5, 22 -- Date: Wed, 7 Feb 2007 07:54:06 -0600 5, 22 -- Organization: University of MN 5, 22 -- Message-ID: {007501c74abf$6f557930$8e105ea0-at-charfac.umn.edu} 5, 22 -- MIME-Version: 1.0 5, 22 -- Content-Type: text/plain; 5, 22 -- charset="US-ASCII" 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Mailer: Microsoft Office Outlook 11 5, 22 -- Thread-Index: AcdFUmwPSTknqTYATeS1IJxxMfwe4w== 5, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
We are advertising for a TEM sample preparation technician at the UMN Characterization Facility. This position's emphasis will be on bio-medical sample prep experience and microtomy rather than prep of hard materials. Please see our ad at the University of MN job site link for details: http://employment.umn.edu/applicants/Central?quickFind=59448
or you can contact me directly for more information.
Mike ****************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 18 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 MiNTeC node of the National Nanotechnology Infrastructure Network
University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://www.charfac.umn.edu ********************************************************************
==============================Original Headers============================== 5, 22 -- From mboucher-at-umn.edu Wed Feb 7 08:20:40 2007 5, 22 -- Received: from mta-a2.tc.umn.edu (mta-a2.tc.umn.edu [134.84.119.206]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17EKdks015014 5, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 08:20:39 -0600 5, 22 -- Received: from mike (Mike.charfac.umn.edu [160.94.16.142]) 5, 22 -- by mta-a2.tc.umn.edu (UMN smtpd) with ESMTP 5, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 08:20:39 -0600 (CST) 5, 22 -- X-Umn-Remote-Mta: [N] Mike.charfac.umn.edu [160.94.16.142] #+LO+TS+AU+HN 5, 22 -- Reply-To: {mboucher-at-umn.edu} 5, 22 -- From: "Michael L. Boucher" {mboucher-at-umn.edu} 5, 22 -- To: {Microscopy-at-microscopy.com} 5, 22 -- Subject: TEM sample preparation technician needed 5, 22 -- Date: Wed, 7 Feb 2007 08:24:47 -0600 5, 22 -- Organization: University of MN 5, 22 -- Message-ID: {007601c74ac3$b8e8c850$8e105ea0-at-charfac.umn.edu} 5, 22 -- MIME-Version: 1.0 5, 22 -- Content-Type: text/plain; 5, 22 -- charset="US-ASCII" 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Mailer: Microsoft Office Outlook 11 5, 22 -- Thread-Index: AcdKw7iVv6KqL17PSZWpr3eL8jCfOg== 5, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
I've also used ethanol as the transfer fluid with liquid CO2 for many years. Both Polaron and Ladd CPD's have been used without any problems. I tried amyl acetate for awhile but wasn't too fond of it. It's only advantage was being able to smell minute residues of it during the CPD flushing process.
However, the number of flushes of liquid CO2 and length of time between flushes depends on the size or thickness of your sample. Most of the ethanol is removed in the first few flushes. You really have to become familar with your samples to determine the time required to remove all of the ethanol. And as usual add a few flushes for good measure.
I've found certain arthrpods/insects etc. to be most difficult using CPD. The hard exoskeleton or cuticle allows ethanol to enter the body of the organism but doesn't easily allow the exchange of ethanol with liquid CO2. Tardigrades were especially aggravating. Alternative drying techniques than CPD with arthropods and insects are welcome.
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Dear David,
I routinely use acetone in a Balzers critical point drier but sometimes want to dry samples that may be damaged by the acetone. In these cases (for example preserving the outer surfaces of insect eggs prior to SEM examination) I use ethanol. From my experience, the ethanol is nowhere near as miscible as acetone in liquid CO2 but still works perfectly well.
You need to use more exchanges of the liquid CO2 in order to replace the ethanol, leaving to stand for a few minutes between each exchange. If your CPD system has a stirrer, even better! I find that the drying process in the CPD with ethanol can take around four times longer than the conventional technique using acetone. However as long as it achieves the result, then the additional time has been worthwhile.
Best of luck!
Regards,
Chris
Chris Jones ----------------------------------- Hitachi High-Technologies 7 Ivanhoe Road Hogwood Industrial Estate Finchampstead Wokingham Berks RG40 4QQ Tel +44(0)118 932 8632 Fax +44(0)118 973 2622 chrisj-at-hitachi-hitec-uk.com www.hitachi-hitec-uk.com
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We have used ethanol followed by liquid carbon dioxide for years in our Tousimis Samdri-780 CPD. I switched from acetone, which I have been told mixes better than ethanol with liquid carbon dioxide, to reduce extraction from samples.
Dave
-----Original Message----- X-from: dlowry-at-asu.edu [mailto:dlowry-at-asu.edu] Sent: 07 February 2007 07:18 To: David Patton
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Email: dlowry-at-asu.edu Name: David Lowry
Organization: Arizona State University
Title-Subject: [Filtered] critical point drying question
Question: Is ethanol miscible in liquid CO2? I can't seem to find anywhere this is explicitly stated. I have seen SEM protocols which imply they use EtOH as dehydrating agent and then directly CPD with CO2. We have an older Balzers CPD020 unit and the manual has a flow-chart which shows only acetone or amyl acetate as dehydrants prior to CO2 transition. It is my understanding that EtOH is a weaker organic solvent than acetone, and if EtOH mixes with CO2, this may be useful for certain CPD applications. Thanks
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC CSQ-EM 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov bingber46-at-hotmail.com 504-286-4270 desk phone 504-782-6323 cell
==============================Original Headers============================== 30, 20 -- From bingber-at-srrc.ars.usda.gov Wed Feb 7 09:08:08 2007 30, 20 -- Received: from srrc.ars.usda.gov (marconi.srrc.ars.usda.gov [199.133.86.11]) 30, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17F85uo027195 30, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Feb 2007 09:08:06 -0600 30, 20 -- Received: from (unknown [199.133.86.11]) by DA32USMOKC1_AVS01.usda.gov with smtp 30, 20 -- id 1204_75cf2180_b6c1_11db_a6c5_001143d22ff1; 30, 20 -- Wed, 07 Feb 2007 15:39:56 +0000 30, 20 -- Received: from SRRCDOM-MTA by srrc.ars.usda.gov 30, 20 -- with Novell_GroupWise; Wed, 07 Feb 2007 09:08:04 -0600 30, 20 -- Message-Id: {s5c996f4.022-at-srrc.ars.usda.gov} 30, 20 -- X-Mailer: Novell GroupWise Internet Agent 6.0.4 30, 20 -- Date: Wed, 07 Feb 2007 09:07:41 -0600 30, 20 -- From: "Bruce Ingber" {bingber-at-srrc.ars.usda.gov} 30, 20 -- To: {Microscopy-at-MSA.Microscopy.Com} 30, 20 -- Subject: [Microscopy] RE: viaWWW: critical point drying question 30, 20 -- Mime-Version: 1.0 30, 20 -- Content-Type: text/plain; charset=US-ASCII 30, 20 -- Content-Disposition: inline 30, 20 -- Content-Transfer-Encoding: 8bit 30, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l17F85uo027195 ==============================End of - Headers==============================
At the risk of being a little commercial, I'd also recommend getting a copy of our book, Optimizing Light MIcroscopy for Biological and Clinical labs. It has over 80 mini experiments that you might find helpful in your classroom.
Hope this was helpful, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through July. Call us today for details.
P. S. Need a good general reference or light microscopy text for the Spring semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.
At 09:24 AM 2/7/2007, miranda.s.norby-at-sendit.nodak.edu wrote:
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==============================Original Headers============================== 13, 17 -- From bfoster-at-mme1.com Wed Feb 7 09:30:18 2007 13, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 13, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17FUG8C006331 13, 17 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 09:30:17 -0600 13, 17 -- Received: (qmail 11426 invoked by uid 2020); 7 Feb 2007 09:58:19 -0600 13, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 13, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 7 Feb 2007 09:58:19 -0600 13, 17 -- Message-Id: {7.0.1.0.0.20070207092847.01df2648-at-mme1.com} 13, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 17 -- Date: Wed, 07 Feb 2007 09:30:08 -0600 13, 17 -- To: miranda.s.norby-at-sendit.nodak.edu, microscopy-at-microscopy.com 13, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 13, 17 -- Subject: Re: [Microscopy] AskAMicroscopist: student microscopes 13, 17 -- In-Reply-To: {200702070715.l177FU3k026215-at-ns.microscopy.com} 13, 17 -- References: {200702070715.l177FU3k026215-at-ns.microscopy.com} 13, 17 -- Mime-Version: 1.0 13, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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Does anyone have any explanation for what is the function of PVP(polyvinylpyrrodine) in the phosphate buffer for fixing perfusion? I have seen it in an article and now I am curious. Thanks in advance.
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Email: martin.roe-at-nottingham.ac.uk Name: Martin Roe
Organization: Nottingham University
Title-Subject: [Filtered] EDX detectors for JEOL 4000FX TEM
Question: Hello listservers, Does anyone have an old/spare high angle EDX detector (preferably a LINK/ OI) for a JEOL 4000FX TEM? Secondly, does anyone happen to know if an EDX detector was ever made suitable for use on the same microscope's horizontal port (on objective lens) i.e. a horizontally mounted detector. Thanks Martin Roe
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Miranda -
Please look at the "buying school microscopes" section of the Project MICRO web page (URL below) for detailed general advice. Since you teach a wide grade range, be sure to get both dissecting and compound scopes. And since this isn't the best of all possible worlds, you'll need to control price. DON'T do that by buying used equipment on eBay; research-grade scopes that get to that marketplace are too complex for young students to use correctly, and they usually have condition problems. Don't buy 100x objectives on the compound scopes; they require immersion oil and an adjustable condenser, which students won't use properly. That said, look for monocular compound scopes with 4-10-40x objectives, fixed condenser & rechargable LED illumination; you'll find remarkably good imports for around $150. Durable monocular dissecting scopes are $70, and there's a great portable scope for field trips for the same price. I'll be happy to look at web listings of your tentative selections, and to answer further questions; since I'm a retiree, I promise to be prompt.
Listers: If you care enough about this topic to have read this far, I've probably horrified some of you. Please visit the MICRO booth at M&M '07 in Ft. Lauderdale to see some of these scopes!
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
==============================Original Headers============================== 5, 18 -- From schooley-at-mcn.org Wed Feb 7 12:24:35 2007 5, 18 -- Received: from dns3.mcn.org (dns3.mcn.org [216.150.240.32]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17IOY5U011652 5, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 12:24:34 -0600 5, 18 -- Received: from [66.52.170.76] 5, 18 -- by dns3.mcn.org with esmtpa (Exim 4.60) 5, 18 -- (envelope-from {schooley-at-mcn.org} ) 5, 18 -- id JD3V4W-0008YS-4J; Wed, 07 Feb 2007 10:24:34 -0800 5, 18 -- Mime-Version: 1.0 5, 18 -- Message-Id: {a06200702c1efbae62ab9-at-[66.52.170.185]} 5, 18 -- In-Reply-To: {200702070719.l177JKik007366-at-ns.microscopy.com} 5, 18 -- References: {200702070719.l177JKik007366-at-ns.microscopy.com} 5, 18 -- Date: Wed, 7 Feb 2007 09:53:05 -0800 5, 18 -- To: miranda.s.norby-at-sendit.nodak.edu 5, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 5, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: student microscopes 5, 18 -- Cc: microscopy-at-microscopy.com, tpepper-at-iastate.edu 5, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Several people have written to me about my comments on floaters and cataracts, and so I gather there is some general interest in this subject. Therefore, I would like to pass along some comments made by Linus Pauling about these matters in his book "How To Live Longer and Feel Better' (ISBN 0-7167-1775-1).
In Chapter 23 he cites research results that make the following points relating to the eyes: The concentration of vitamin C in the aqueous humor of the eye is twenty five times as high as in blood plasma (suggesting that taking significant amounts of C might be beneficial to the quality of this fluid) Many investigators have reported that there is very little vitamin C in the aqueous humor of eyes that have cataracts, and that patients with them usually have low concentrations in their blood plasma. High intakes of vitamins C and E and B12 appear to be beneficial in preventing senile cataracts. Regular intakes of high doses of vitamins C also appears to be beneficial in preventing and controlling glaucoma.
Pauling doesn't mention floaters specifically, but it would seem reasonable to expect that anything that would improve the quality of the fluid in the eye might also reduce the occurrence of these pesky inclusions.
Pauling recommends a daily intake of 6 to 18 gm of vitamin C, 400 to 1600 IU of vitamin E, and one or two Super-B 50 tablets, plus a mineral supplement. I haven't taken vitamins at the full level he recommends, but have been taking 2 gm of C, 400 IU of E, and one Super-B 50, plus a theraputic vitamin+mineral tablet, daily for the past 20 years and think doing so has been very beneficial in many ways.
-- 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============================== 5, 15 -- From bigelow-at-engin.umich.edu Wed Feb 7 14:55:51 2007 5, 15 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 5, 15 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17Ktp7S004633 5, 15 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 14:55:51 -0600 5, 15 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 5, 15 -- by srvr22.engin.umich.edu (8.13.6/8.13.6) with ESMTP id l17Kto4I009190; 5, 15 -- Wed, 7 Feb 2007 15:55:50 -0500 (EST) 5, 15 -- Mime-Version: 1.0 5, 15 -- Message-Id: {p06210201c1efe79378ce-at-[141.212.131.221]} 5, 15 -- Date: Wed, 7 Feb 2007 15:55:49 -0500 5, 15 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 5, 15 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 5, 15 -- Subject: [Microscopy] RE: Floaters and cataracts 5, 15 -- Cc: MSE Faculty {mse-faculty-at-umich.edu} , mse-office-staff-at-umich.edu 5, 15 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Does anyone have any experience cleaning the carbonaeous gunk off of Mo or Pt-Ir apertures with a solvent? I know about heating up in a Mo etc boat, but looking for alternatives. Will ammonium hydroxide do the trick? Or something else?
thanks. -- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
==============================Original Headers============================== 4, 24 -- From johnf-at-geology.wisc.edu Wed Feb 7 15:07:22 2007 4, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17L7MvG015890 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:22 -0600 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by localhost (Postfix) with ESMTP id 9A3E520D20 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:21 -0600 (CST) 4, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 4, 24 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 4, 24 -- with ESMTP id 07529-02 for {microscopy-at-microscopy.com} ; 4, 24 -- Wed, 7 Feb 2007 15:07:14 -0600 (CST) 4, 24 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 4, 24 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 4, 24 -- (No client certificate requested) 4, 24 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 29B0A20D02 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:14 -0600 (CST) 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {p06230910c1efee78374f-at-[144.92.206.57]} 4, 24 -- Date: Wed, 7 Feb 2007 15:03:05 -0600 4, 24 -- To: microscopy-at-microscopy.com 4, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 4, 24 -- Subject: cleaning apertures chemically? 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
John, I've had very good luck for the last 30 years or so cleaning apertures with a cut knap polishing cloth and 1 micron diamond paste. Just place the aperture on the cloth with a little paste and put your finger on it and rub it in a circular motion. Do both sides. Clean ultrasonically in Joy dishwashing liquid and hot water, rinse in hot tap water or distilled water and immediately blow dry with a duster to avoid water spots. My understanding about Joy is that the Proctor & Gamble labs use it for cleaning critical parts of AAUs and can find no residue. Apparently this is not true of all dishwashing liquids.
This works with 1 mil foil (including multi-hole strips), 5 mil countersunk and even the little Siemems apertures that are heavily countersunk. Just don't try it with gold foil self-cleaning apertures. The gold foil is far too thin and fragile for this technique.
Chuck Garber at SPI tells me that most metallographic diamond pastes have silicones in them. That's a problem, but his pastes don't. As a bonus, the SPI prices are very good. No financial interest, just a happy user.
As you are probably aware, heating moly in a platinum boat is a problem and even heating Pt has limited usefulness because the Pt recrystalizes and eventually you have an aperture with alligator skin and it won't work any more (high astigmatism). Polishing a ruined Pt aperture will restore it, probably due to the smearing of the metal by the diamond particles.
All in all it's pretty good because you don't need to know what the aperture is made of, there's no complicated or exotic equipment needed (beyond an ultrasonic cleaner), no organic solvents or other nasty stuff, and you can use the same apertures for years. Once in a great while I might fold a 1 mil aperture, but that doesn't happen very often.
Ken Converse Owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: johnf-at-geology.wisc.edu [mailto:johnf-at-geology.wisc.edu] Sent: Wednesday, February 07, 2007 4:11 PM To: kenconverse-at-qualityimages.biz
Does anyone have any experience cleaning the carbonaeous gunk off of Mo or Pt-Ir apertures with a solvent? I know about heating up in a Mo etc boat, but looking for alternatives. Will ammonium hydroxide do the trick? Or something else?
thanks. -- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
==============================Original Headers============================== 4, 24 -- From johnf-at-geology.wisc.edu Wed Feb 7 15:07:22 2007 4, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l17L7MvG015890 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:22 -0600 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by localhost (Postfix) with ESMTP id 9A3E520D20 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:21 -0600 (CST) 4, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 4, 24 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 4, 24 -- with ESMTP id 07529-02 for {microscopy-at-microscopy.com} ; 4, 24 -- Wed, 7 Feb 2007 15:07:14 -0600 (CST) 4, 24 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 4, 24 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 4, 24 -- (No client certificate requested) 4, 24 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 29B0A20D02 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:14 -0600 (CST) 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {p06230910c1efee78374f-at-[144.92.206.57]} 4, 24 -- Date: Wed, 7 Feb 2007 15:03:05 -0600 4, 24 -- To: microscopy-at-microscopy.com 4, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 4, 24 -- Subject: cleaning apertures chemically? 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
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==============================Original Headers============================== 25, 29 -- From kenconverse-at-qualityimages.biz Wed Feb 7 21:03:14 2007 25, 29 -- Received: from dpmailmta02.doteasy.com (dpmailmta02.doteasy.com [65.61.209.81]) 25, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1833EcY005037 25, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Feb 2007 21:03:14 -0600 25, 29 -- Received: from qualityimages.biz (unverified [192.168.101.16]) 25, 29 -- by dpmailmta02.doteasy.com (DEO) with ESMTP id 57086489-1814644 25, 29 -- for multiple; Wed, 07 Feb 2007 19:43:42 -0800 25, 29 -- Received: from Ken [68.193.55.199] by qualityimages.biz with ESMTP 25, 29 -- (SMTPD32-8.05) id A2EEE2DE004C; Wed, 07 Feb 2007 19:03:10 -0800 25, 29 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 25, 29 -- To: {johnf-at-geology.wisc.edu} 25, 29 -- Cc: "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 25, 29 -- Subject: RE: [Microscopy] cleaning apertures chemically? 25, 29 -- Date: Wed, 7 Feb 2007 22:02:53 -0500 25, 29 -- Message-ID: {006501c74b2d$a21cb3b0$8d7ec44b-at-Ken} 25, 29 -- MIME-Version: 1.0 25, 29 -- Content-Type: text/plain; 25, 29 -- charset="us-ascii" 25, 29 -- X-Priority: 3 (Normal) 25, 29 -- X-MSMail-Priority: Normal 25, 29 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 25, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 25, 29 -- In-Reply-To: {200702072111.l17LB5uK020615-at-ns.microscopy.com} 25, 29 -- Importance: Normal 25, 29 -- X-IMSTrailer: __IMail_7__ 25, 29 -- X-IP-stats: Incoming Last 0, First 133, in=3174090, out=0, spam=0 ip=192.168.101.16 25, 29 -- X-External-IP: 192.168.101.16 25, 29 -- Content-Transfer-Encoding: 8bit 25, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1833EcY005037 ==============================End of - Headers==============================
Let me float a question out into E-space about microchemical/spot test. My limited experience suggest that high levels of sulfate (SO4) interferes or reduces the sensitivity with Chamot's test for iron. Specifically the potassium mercuric thiocyanate, potassium thiocyanate and the potassium ferrocyanide test. Has anyone else noticed this problem?
I solved it by repetitively dry boiling my sample in dilute (15%) HNO3. I'm just looking for confirmation.......
Stay safe...................Frank
==============================Original Headers============================== 4, 17 -- From frank.karl-at-degussa.com Thu Feb 8 08:21:22 2007 4, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18ELLTm029160 4, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Feb 2007 08:21:22 -0600 4, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 4, 17 -- by malmailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with SMTP id l18ELGjs007029 4, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Feb 2007 15:21:19 +0100 4, 17 -- Subject: Question about sulfate interference in FE spot test 4, 17 -- To: microscopy-at-msa.microscopy.com 4, 17 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 4, 17 -- Message-ID: {OF99FC0C3D.283ECD2B-ON8625727C.004DF726-8525727C.004ED802-at-degussa.com} 4, 17 -- From: frank.karl-at-degussa.com 4, 17 -- Date: Thu, 8 Feb 2007 09:21:11 -0500 4, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 4, 17 -- 02/08/2007 08:21:20 AM 4, 17 -- MIME-Version: 1.0 4, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I have just recently begun using the Leica EM FCS cryo set-up for our ultramicrotome. I'm having a little difficulty and was hoping that members of this list could clue me into some tricks of the trade. I am presently trying to section polymer samples and I need them to be around 50nm in thickness. During cutting, the sections often curl or fold like an accordion. So I pick them up with an eyelash tool and try (with much difficulty and cold fingers) to unfold and stretch them out onto a copper grid. The sections have no affinity for the Cu grid and are much happier adhering to the eyelash or wadding up into an unusable lump. Short sections don't work any better, since these just tend to flip away. I'm using a anti-static device but it's difficult to place it into the correct position since I'm using it's holder to hold the copper grids close to the diamond.
Thank you for any advice! Shawn
==============================Original Headers============================== 3, 30 -- From trent-at-ornl.gov Thu Feb 8 10:04:13 2007 3, 30 -- Received: from emroute3.ornl.gov (emroute3.ornl.gov [160.91.4.110]) 3, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18G4DcX010021 3, 30 -- for {Microscopy-at-Microscopy.Com} ; Thu, 8 Feb 2007 10:04:13 -0600 3, 30 -- Received: from emroute3.ornl.gov ([127.0.0.1]) 3, 30 -- by emroute3.ornl.gov (PMDF V6.3-x3 #31246) 3, 30 -- with ESMTP id {0JD500FGMJ4X97-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from CONVERSION-DAEMON.emroute3.ornl.gov by emroute3.ornl.gov 3, 30 -- (PMDF V6.3-x3 #31246) id {0JD500G01J4XX8-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from ORNLEXCHANGE.ornl.gov (ornlexchange1.ornl.gov [160.91.1.20]) 3, 30 -- by emroute3.ornl.gov (PMDF V6.3-x3 #31246) 3, 30 -- with ESMTP id {0JD500FBBJ4X9V-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from 160.91.157.36 ([160.91.157.36]) 3, 30 -- by ORNLEXCHANGE.ornl.gov ([160.91.1.32]) via Exchange Front-End Server 3, 30 -- mail.ornl.gov ([160.91.4.23]) with Microsoft Exchange Server HTTP-DAV ; Thu, 3, 30 -- 08 Feb 2007 16:00:10 +0000 3, 30 -- Date: Thu, 08 Feb 2007 11:00:30 -0500 3, 30 -- From: Shawn Reeves {trent-at-ornl.gov} 3, 30 -- Subject: Cryo ultramicrotomy -help! 3, 30 -- To: Microscopy-at-Microscopy.Com 3, 30 -- Message-id: {C1F0B34E.1804%trent-at-ornl.gov} 3, 30 -- MIME-version: 1.0 3, 30 -- Content-type: text/plain; charset=US-ASCII 3, 30 -- Content-transfer-encoding: 7bit 3, 30 -- User-Agent: Microsoft-Entourage/11.3.3.061214 3, 30 -- Thread-Topic: Cryo ultramicrotomy -help! 3, 30 -- Thread-Index: AcdLmkGKgGPTdreNEduLiwAKlebEPA== ==============================End of - Headers==============================
Are there any technical difficulties with using a side mounted digital camera system versus a bottom mounted camera system for acquiring a tilt series of images for 3D-reconstruction? Please respond privately.
Tom Bargar University of Nebraska Medical Center Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395 402-559-7347 tbargar-at-unmc.edu
==============================Original Headers============================== 4, 20 -- From tbargar-at-unmc.edu Thu Feb 8 10:42:53 2007 4, 20 -- Received: from zixvpm01.unmc.edu (zixvpm01.unmc.edu [192.198.54.126]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18GgrPe000491 4, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 8 Feb 2007 10:42:53 -0600 4, 20 -- Received: from zixvpm01.unmc.edu (ZixVPM [127.0.0.1]) 4, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 6A1CC4C0E3 4, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 8 Feb 2007 10:42:52 -0600 (CST) 4, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 4, 20 -- by zixvpm01.unmc.edu (Proprietary) with ESMTP id 19FBB4C07C 4, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 8 Feb 2007 10:42:51 -0600 (CST) 4, 20 -- Subject: Digital cameras used in electron tomography 4, 20 -- To: Microscopy-at-MSA.Microscopy.com 4, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 4, 20 -- Message-ID: {OF87E25AEB.3DCAE967-ON8625727C.005B5034-8625727C.005BCFB4-at-unmc.edu} 4, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 4, 20 -- Date: Thu, 8 Feb 2007 10:42:49 -0600 4, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 02/08/2007 10:42:51 4, 20 -- AM 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
We are working toward acquiring a 120Kv TEM with electron tomography capability. I would like to hear from anyone who has an instrument (of any brand) at this level. I would like to know your experiences with whatever manufacturer's system you chose. Please, respond privately.
Tom Bargar University of Nebraska Medical Center Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395 402-559-7347 tbargar-at-unmc.edu
==============================Original Headers============================== 4, 20 -- From tbargar-at-unmc.edu Thu Feb 8 10:54:06 2007 4, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18Gs5vC011733 4, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 8 Feb 2007 10:54:06 -0600 4, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 4, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 02FAAA005F 4, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 8 Feb 2007 10:54:04 -0600 (CST) 4, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 4, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 1DD67398047 4, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 8 Feb 2007 10:54:02 -0600 (CST) 4, 20 -- Subject: 120Kv TEM's used in electron tomography 4, 20 -- To: Microscopy-at-MSA.Microscopy.com 4, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 4, 20 -- Message-ID: {OF04C4B81D.1445144E-ON8625727C.005C24D5-8625727C.005CD5FA-at-unmc.edu} 4, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 4, 20 -- Date: Thu, 8 Feb 2007 10:54:00 -0600 4, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 02/08/2007 10:54:01 4, 20 -- AM 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
We are clearing rooms for a couple new scope installations and one gem we turned up is a boxed set of filmstrips and cassette tapes by Crang and Ward, entitled "Electron Microscopy: Principles and Practices", copyright 1975 by the W.B. Saunders Company, Philadelphia, London, Toronto. The box shows some wear, but the filmstrips and cassettes appear to be complete and pristine, with their orignal patina. Some color fade due to age.
Topics covered are: 1) The TEM, 2) Fixation and Embedment of Biological Tissues, 3) Ultramicrotomy, 4) Ultrastructural Localization of Enzime Activity, 5) Electron Microscope Autoradiography, 6) Freeze-etching, 7) The High Voltage Electron Microscope, 8) The SEM, 9) Specimen Preparation for SEM, and 10) X-ray Microanalysis.
At auction, a conservative estimate of value would be whatever emotional attachment someone might attach to them. That's a conservative estimate. Actual value could go even higher.
Any takers for cost of shipping?
Cheers, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 8, 23 -- From TindallR-at-missouri.edu Thu Feb 8 10:57:39 2007 8, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18Gvcv2015610 8, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Feb 2007 10:57:39 -0600 8, 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); 8, 23 -- Thu, 8 Feb 2007 10:57:35 -0600 8, 23 -- x-mimeole: Produced By Microsoft Exchange V6.5 8, 23 -- Content-class: urn:content-classes:message 8, 23 -- MIME-Version: 1.0 8, 23 -- Content-Type: text/plain; 8, 23 -- charset="us-ascii" 8, 23 -- Subject: Electron Microscopy Antique Road Show 8, 23 -- Date: Thu, 8 Feb 2007 10:57:35 -0600 8, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68DCE-at-UM-XMAIL08.um.umsystem.edu} 8, 23 -- X-MS-Has-Attach: 8, 23 -- X-MS-TNEF-Correlator: 8, 23 -- Thread-Topic: Electron Microscopy Antique Road Show 8, 23 -- Thread-Index: AcdLojtPw12MT9HfTee0932OIhsJSw== 8, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 8, 23 -- To: {microscopy-at-microscopy.com} 8, 23 -- X-OriginalArrivalTime: 08 Feb 2007 16:57:35.0530 (UTC) FILETIME=[3B5108A0:01C74BA2] 8, 23 -- Content-Transfer-Encoding: 8bit 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l18Gvcv2015610 ==============================End of - Headers==============================
I would be interested in the feedback on this. Could people post to the listserver? Kim
tbargar-at-unmc.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 } ---------------------------------------------------------------------------- } } } Are there any technical difficulties with using a side mounted digital } camera system versus a bottom mounted camera system for acquiring a tilt } series of images for 3D-reconstruction? Please respond privately. } } Tom Bargar } University of Nebraska Medical Center } Core Electron Microscopy Research Facility } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } 402-559-7347 } tbargar-at-unmc.edu } }
}
-- Kim Rensing Ph.D. Research Assistant Professor Manager, Microscopy and Imaging Facility
University of Calgary Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1 403-220-3488 krensing-at-ucalgary.ca
==============================Original Headers============================== 5, 22 -- From krensing-at-ucalgary.ca Thu Feb 8 11:01:25 2007 5, 22 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18H1PP8027290 5, 22 -- for {microscopy-at-microscopy.com} ; Thu, 8 Feb 2007 11:01:25 -0600 5, 22 -- Received: from [136.159.164.163] (unknown [136.159.164.163]) 5, 22 -- by smtp2.ucalgary.ca (Postfix) with ESMTP id EE78310043 5, 22 -- for {microscopy-at-microscopy.com} ; Thu, 8 Feb 2007 10:01:23 -0700 (MST) 5, 22 -- Message-ID: {45CB5763.7060302-at-ucalgary.ca} 5, 22 -- Date: Thu, 08 Feb 2007 10:01:23 -0700 5, 22 -- From: Kim Rensing {krensing-at-ucalgary.ca} 5, 22 -- Organization: Microscopy and Imaging Facility, U. of Calgary 5, 22 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) 5, 22 -- MIME-Version: 1.0 5, 22 -- To: microscopy-at-microscopy.com 5, 22 -- Subject: Re: [Microscopy] Digital cameras used in electron tomography 5, 22 -- References: {200702081647.l18Glggp010013-at-ns.microscopy.com} 5, 22 -- In-Reply-To: {200702081647.l18Glggp010013-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 5, 22 -- X-UCalgary-MailScanner: Found to be clean 5, 22 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
You can get folding grids that look like two regular mesh grids attached along one side, which is the "hinge". You collect a section on one side, then fold other side over on top so section is clamped in place mechanically, so no adhesion of section to grid takes place. This would also tend to flatten the section to some extent.
Most EM vendors that sell grids would have them.
Whether these folding grids are compatible with requirements for collecting and observing cryosections, I don't know, but it might be worth a try.
Just my 2.5 cents worth...
Good luck,
Gib Gilbert (Gib) Ahlstrand, Electron Microscopist, Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
trent-at-ornl.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 } ---------------------------------------------------------------------------- } } I have just recently begun using the Leica EM FCS cryo set-up for our } ultramicrotome. I'm having a little difficulty and was hoping that members } of this list could clue me into some tricks of the trade. } I am presently trying to section polymer samples and I need them to be } around 50nm in thickness. During cutting, the sections often curl or fold } like an accordion. So I pick them up with an eyelash tool and try (with much } difficulty and cold fingers) to unfold and stretch them out onto a copper } grid. The sections have no affinity for the Cu grid and are much happier } adhering to the eyelash or wadding up into an unusable lump. Short sections } don't work any better, since these just tend to flip away. I'm using a } anti-static device but it's difficult to place it into the correct position } since I'm using it's holder to hold the copper grids close to the diamond. } } Thank you for any advice! } Shawn --
==============================Original Headers============================== 9, 21 -- From ahlst007-at-umn.edu Thu Feb 8 11:42:11 2007 9, 21 -- Received: from mta-a2.tc.umn.edu (mta-a2.tc.umn.edu [134.84.119.206]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18HgAKB013489 9, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 8 Feb 2007 11:42:10 -0600 9, 21 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) 9, 21 -- by mta-a2.tc.umn.edu (UMN smtpd) with ESMTP 9, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 8 Feb 2007 11:42:10 -0600 (CST) 9, 21 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 9, 21 -- Message-ID: {45CB609F.5040402-at-umn.edu} 9, 21 -- Date: Thu, 08 Feb 2007 11:40:47 -0600 9, 21 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 9, 21 -- Reply-To: ahlst007-at-umn.edu 9, 21 -- Organization: Imaging Center UM 9, 21 -- User-Agent: Thunderbird 1.5.0.9 (Macintosh/20061207) 9, 21 -- MIME-Version: 1.0 9, 21 -- To: Microscopy-at-Microscopy.com 9, 21 -- Subject: Re: Cryo ultramicrotomy -help! 9, 21 -- References: {200702081608.l18G8Ohb015860-at-ns.microscopy.com} 9, 21 -- In-Reply-To: {200702081608.l18G8Ohb015860-at-ns.microscopy.com} 9, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both echinea-at-gmx.net as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: echinea-at-gmx.net Name: Ernesto Chinea
Title-Subject: [Filtered] Fixation of whole mosquitoes for SEM and X-ray microscopy
Question: Does anyone have any suggestions for fixation of whole mosquitoes? We are having problems with the preservation of some external structures, particularly the antennas. Is there some ìtrickî to prevent the crumpling of the antennas and the wings? What to do to minimize the lost of limbs? We are using the following protocol: -prefixing in cacodylate buffered GTA at 4 Celsius (no stirring) -gentle buffer washes -fixation in buffered OsO4 -dehydration in graded QUETOL 651 followed by infiltration with QUETOL
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both sswaffe-at-abv.bg as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: V. Andreev
Organization: NPMG
Title-Subject: [Filtered] Comparison
Question: What are the advantages and disadvantages of confocal microscopy in comparison with conventional fluorescence microscopy? For example the level of complexity when it comes to manipulating the microscope and specimen processing, hazardous issues, image quality and diversity in possible techniques. Which one of the two systems is more used in clinical or research areas?
On Feb 8, 2007, at 8:42 AM, tbargar-at-unmc.edu wrote:
} Are there any technical difficulties with using a side mounted digital } camera system versus a bottom mounted camera system for acquiring a } tilt } series of images for 3D-reconstruction? Please respond privately. } Dear Tom, We have taken tilt series on our T12 with a camera that inserts from the side of the column and there were no difficulties. If you refer to an off-axis camera, which uses image shifts to put the image onto the camera, I could foresee some potential problems, but I would expect that the software would handle everything properly. If you are using low-dose conditions, the camera has to have (the option of) a pre-specimen shutter, but other than that, the data collection should go the same way regardless of where the camera is mounted. 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, 24 -- From tivol-at-caltech.edu Thu Feb 8 12:32:16 2007 4, 24 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18IWGW5015542 4, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Feb 2007 12:32:16 -0600 4, 24 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 24 -- by fire-ox-postvirus (Postfix) with ESMTP id A93BA3569C 4, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Feb 2007 10:32:15 -0800 (PST) 4, 24 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 24 -- (using TLSv1 with cipher RC4-SHA (128/128 bits)) 4, 24 -- (No client certificate requested) 4, 24 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 3684510A170 4, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Feb 2007 10:32:12 -0800 (PST) 4, 24 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 24 -- In-Reply-To: {200702081642.l18Ggxhu000651-at-ns.microscopy.com} 4, 24 -- References: {200702081642.l18Ggxhu000651-at-ns.microscopy.com} 4, 24 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 24 -- Message-Id: {50b4a08af32f65c9847f828026a947c8-at-caltech.edu} 4, 24 -- Content-Transfer-Encoding: 7bit 4, 24 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 24 -- Subject: Re: [Microscopy] Digital cameras used in electron tomography 4, 24 -- Date: Thu, 8 Feb 2007 10:40:35 -0800 4, 24 -- To: microscopy-at-msa.microscopy.com 4, 24 -- X-Mailer: Apple Mail (2.624) 4, 24 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Hi I suggest cutting the mosquito into 2-3 pieces (in the prefixative) with a sharp razor blade (e.g. head, thorax and abdomen?). This should improve fixation and embedding. The cuticle is usually very hard to penetrate and cutting the insect will hopefully improve permeability of tissues to all used reagents. I have done this with leafhoppers and aphids with normally good results. Good luck.
The filmstrips were claimed within minutes after the posting appeared, but thanks for all the interest. I didn't know so many people were interested in these old things. Now, let's see what else I can find...........
Cheers, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 5, 23 -- From TindallR-at-missouri.edu Thu Feb 8 13:02:39 2007 5, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18J2dSM016735 5, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Feb 2007 13:02:39 -0600 5, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 5, 23 -- Thu, 8 Feb 2007 13:02:38 -0600 5, 23 -- x-mimeole: Produced By Microsoft Exchange V6.5 5, 23 -- Content-class: urn:content-classes:message 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; 5, 23 -- charset="us-ascii" 5, 23 -- Subject: EM Antiques 5, 23 -- Date: Thu, 8 Feb 2007 13:02:38 -0600 5, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68DD2-at-UM-XMAIL08.um.umsystem.edu} 5, 23 -- X-MS-Has-Attach: 5, 23 -- X-MS-TNEF-Correlator: 5, 23 -- Thread-Topic: EM Antiques 5, 23 -- Thread-Index: AcdLs7Nm2ijlb3vmQ4Sg30eSLdG4Gw== 5, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- X-OriginalArrivalTime: 08 Feb 2007 19:02:38.0962 (UTC) FILETIME=[B3B5E520:01C74BB3] 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l18J2dSM016735 ==============================End of - Headers==============================
are you cutting dry or do you float the sections? In case you are cutting wet, it helps to use a pipette filled with the water/DMSO mixture at room temperature and bring some of the "hot" liquid under the folded sections. The fluctuations in the fluid caused by the temperature difference will unfold the cuts very nicely. Hope this helps
Petra --------------------------------------- Dr. Petra Wahlbring Goodyear S.A. Technical Center Analytical Test Laboratories L-7750 Colmar-Berg Luxembourg e-mail: petra.wahlbring-at-goodyear.com
- May Contain Confidential and/or Proprietary Information. May not be copied or disseminated without the expressed written consent of The Goodyear Tire & Rubber Company. -
trent-at-ornl.gov
02/08/07 05:08 PM To petra.wahlbring-at-goodyear.com cc Please respond to trent-at-ornl.gov Subject [Microscopy] Cryo ultramicrotomy -help!
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
I have just recently begun using the Leica EM FCS cryo set-up for our ultramicrotome. I'm having a little difficulty and was hoping that members of this list could clue me into some tricks of the trade. I am presently trying to section polymer samples and I need them to be around 50nm in thickness. During cutting, the sections often curl or fold like an accordion. So I pick them up with an eyelash tool and try (with much difficulty and cold fingers) to unfold and stretch them out onto a copper grid. The sections have no affinity for the Cu grid and are much happier adhering to the eyelash or wadding up into an unusable lump. Short sections don't work any better, since these just tend to flip away. I'm using a anti-static device but it's difficult to place it into the correct position since I'm using it's holder to hold the copper grids close to the diamond.
Thank you for any advice! Shawn
==============================Original Headers============================== 3, 30 -- From trent-at-ornl.gov Thu Feb 8 10:04:13 2007 3, 30 -- Received: from emroute3.ornl.gov (emroute3.ornl.gov [160.91.4.110]) 3, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18G4DcX010021 3, 30 -- for {Microscopy-at-Microscopy.Com} ; Thu, 8 Feb 2007 10:04:13 -0600 3, 30 -- Received: from emroute3.ornl.gov ([127.0.0.1]) 3, 30 -- by emroute3.ornl.gov (PMDF V6.3-x3 #31246) 3, 30 -- with ESMTP id {0JD500FGMJ4X97-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from CONVERSION-DAEMON.emroute3.ornl.gov by emroute3.ornl.gov 3, 30 -- (PMDF V6.3-x3 #31246) id {0JD500G01J4XX8-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from ORNLEXCHANGE.ornl.gov (ornlexchange1.ornl.gov [160.91.1.20]) 3, 30 -- by emroute3.ornl.gov (PMDF V6.3-x3 #31246) 3, 30 -- with ESMTP id {0JD500FBBJ4X9V-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from 160.91.157.36 ([160.91.157.36]) 3, 30 -- by ORNLEXCHANGE.ornl.gov ([160.91.1.32]) via Exchange Front-End Server 3, 30 -- mail.ornl.gov ([160.91.4.23]) with Microsoft Exchange Server HTTP-DAV ; Thu, 3, 30 -- 08 Feb 2007 16:00:10 +0000 3, 30 -- Date: Thu, 08 Feb 2007 11:00:30 -0500 3, 30 -- From: Shawn Reeves {trent-at-ornl.gov} 3, 30 -- Subject: Cryo ultramicrotomy -help! 3, 30 -- To: Microscopy-at-Microscopy.Com 3, 30 -- Message-id: {C1F0B34E.1804%trent-at-ornl.gov} 3, 30 -- MIME-version: 1.0 3, 30 -- Content-type: text/plain; charset=US-ASCII 3, 30 -- Content-transfer-encoding: 7bit 3, 30 -- User-Agent: Microsoft-Entourage/11.3.3.061214 3, 30 -- Thread-Topic: Cryo ultramicrotomy -help! 3, 30 -- Thread-Index: AcdLmkGKgGPTdreNEduLiwAKlebEPA== ==============================End of - Headers==============================
==============================Original Headers============================== 22, 16 -- From petra.wahlbring-at-goodyear.com Fri Feb 9 01:55:52 2007 22, 16 -- Received: from emeam1.goodyear.com (emeam1.goodyear.com [57.67.226.7]) 22, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l197tqYJ014525 22, 16 -- for {Microscopy-at-Microscopy.Com} ; Fri, 9 Feb 2007 01:55:52 -0600 22, 16 -- In-Reply-To: {200702081608.l18G8uot016707-at-ns.microscopy.com} 22, 16 -- Subject: Re: [Microscopy] Cryo ultramicrotomy -help! 22, 16 -- To: trent-at-ornl.gov 22, 16 -- Cc: Microscopy-at-Microscopy.Com 22, 16 -- X-Mailer: Lotus Notes Release 6.5 September 26, 2003 22, 16 -- Message-ID: {OF2115C3C7.24FEE0BC-ONC125727D.002AA858-C125727D.002B5A21-at-goodyear.com} 22, 16 -- From: petra.wahlbring-at-goodyear.com 22, 16 -- Date: Fri, 9 Feb 2007 08:53:30 +0100 22, 16 -- X-MIMETrack: Serialize by Router on ECLNGM01/EU/GDYRNET(Release 6.5.4FP3|January 09, 2006) at 22, 16 -- 02/09/2007 08:55:52 AM 22, 16 -- MIME-Version: 1.0 22, 16 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
OK, I've had enough. Has anybody come up with a way to export all the spectra in an Oxford Inca Project (to EMSA format in particular) in one fell swoop? I'm getting a little tired the right click fandango for each and every spectra. If it matters, I have Inca 200 version 4.02.
My worst nightmare: Sure, version x.xx can do that. It's only a zillion dollars. I don't need any more pretty graphics and hand holding. I'd just like to get mundane grunt work done fast. After all, that's what computers are for.....
At least it's Friday.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
If Photoshop is used to modify a digital TEM image to remove stain precipitate, how should the image processing be described in the figure legend......or is it even ethical to modify an image in this way?
Thank you in advance for your opinions!
-- Sandy Hancock Laboratory for Neurotoxicity Studies Virginia-Maryland Regional College of Veterinary Medicine Virginia Tech (540)231-4817
==============================Original Headers============================== 5, 18 -- From hancocksk-at-vt.edu Fri Feb 9 10:04:04 2007 5, 18 -- Received: from lennier.cc.vt.edu (lennier.cc.vt.edu [198.82.162.213]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19G44oT014138 5, 18 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 10:04:04 -0600 5, 18 -- Received: from vivi.cc.vt.edu (IDENT:mirapoint-at-evil-vivi.cc.vt.edu [10.1.1.12]) 5, 18 -- by lennier.cc.vt.edu (8.12.11.20060308/8.12.11) with ESMTP id l19Fv54C026240 5, 18 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 11:04:17 -0500 5, 18 -- Received: from [128.173.229.225] (vet9225.vetmed.vt.edu [128.173.229.225]) 5, 18 -- by vivi.cc.vt.edu (MOS 3.8.2-GA) 5, 18 -- with ESMTP id GZH72300; 5, 18 -- Fri, 9 Feb 2007 11:04:03 -0500 (EST) 5, 18 -- Mime-Version: 1.0 5, 18 -- Message-Id: {a06230987c1f24bdeeb1c-at-[128.173.229.225]} 5, 18 -- Date: Fri, 9 Feb 2007 11:04:00 -0500 5, 18 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 5, 18 -- From: Sandy Hancock {skperkin-at-vt.edu} 5, 18 -- Subject: image processing 5, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
The interdisciplinary meeting on Electron Microscopy and Multiscale Modelling (EMMM 2007) will take place in Moscow, Russian Federation, from 3-7 September 2007. This meeting, which is intended to bring together experts from several key areas of modern crystallography and materials science, will focus on the link between methods of direct visualization of structures at nano- and meso-scale based on diffraction techniques and electron microscopy, and the rapidly expanding field of multiscale modelling of materials.
Further information about the meeting, including the provisional programme, information about the city of Moscow, transportation and accommodation, is available at: http://www.crys.ras.ru/EMMM07/
-- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2220 N Campus Drive Northwestern University Evanston, IL 60208, USA Tel: (847) 491-3996 Fax: (847) 491-7820 email: L-marks at northwestern dot edu www.numis.northwestern.edu EMM2007 http://ns.crys.ras.ru/EMMM07/
==============================Original Headers============================== 3, 22 -- From marksmsa-at-gmail.com Fri Feb 9 10:04:35 2007 3, 22 -- Received: from ug-out-1314.google.com (ug-out-1314.google.com [66.249.92.174]) 3, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19G4Y1I014925 3, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 10:04:35 -0600 3, 22 -- Received: by ug-out-1314.google.com with SMTP id m2so772354ugc 3, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 09 Feb 2007 08:04:33 -0800 (PST) 3, 22 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 3, 22 -- d=gmail.com; s=beta; 3, 22 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 3, 22 -- b=IPaL4LyBQdxPp3V68f3gCQ4Q1/pfS4add0SjHQR3EeyKX6tnXcj0EupubcS3rxXlU0D5peZBs8DPPkE6olrqFSQMZiHQ0kGqPuPWfR7CJVKDC9L/pdxmYVqx1UkvSsJYoZTFv9DDQaMZGyZDs7HrXlXTQ+O74uP4ZUeOJph5icc= 3, 22 -- Received: by 10.78.39.16 with SMTP id m16mr5013467hum.1171037072108; 3, 22 -- Fri, 09 Feb 2007 08:04:32 -0800 (PST) 3, 22 -- Received: by 10.78.42.1 with HTTP; Fri, 9 Feb 2007 08:04:32 -0800 (PST) 3, 22 -- Message-ID: {e13ba6260702090804m2776f164ud96debfe924ecb9d-at-mail.gmail.com} 3, 22 -- Date: Fri, 9 Feb 2007 10:04:32 -0600 3, 22 -- From: "L Marks" {marksmsa-at-gmail.com} 3, 22 -- To: Microscopy-at-microscopy.com 3, 22 -- Subject: EMMM 2007: First Announcement 3, 22 -- MIME-Version: 1.0 3, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 22 -- Content-Transfer-Encoding: 7bit 3, 22 -- Content-Disposition: inline ==============================End of - Headers==============================
If you are cutting dry, maybe using the technique we use for biological samples (Tokuyasu wet retrieval method). Pick your section up using a drop of 2.3M sucrose in phosphate buffer on a small 2mm loop. Dip the loop in the sucrose and then bring the drop over the section and lower it until the section "sticks" to the bottom of the drop. The sucrose will freeze quickly, so be fast. Remove the loop to room temp and wait a couple seconds for the droplet to melt and then touch it to your prepared grid. The section will attach to the grid and hopefully be flat again. Again, the temperature difference may help straighten them out. Good luck, Jo Dee
Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease
Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158
-----Original Message----- X-from: petra.wahlbring-at-goodyear.com [mailto:petra.wahlbring-at-goodyear.com] Sent: Friday, February 09, 2007 12:02 AM To: jfish-at-gladstone.ucsf.edu
Dear Shawn,
are you cutting dry or do you float the sections? In case you are cutting wet, it helps to use a pipette filled with the water/DMSO mixture at room temperature and bring some of the "hot" liquid under the folded sections. The fluctuations in the fluid caused by the temperature difference will unfold the cuts very nicely. Hope this helps
Petra --------------------------------------- Dr. Petra Wahlbring Goodyear S.A. Technical Center Analytical Test Laboratories L-7750 Colmar-Berg Luxembourg e-mail: petra.wahlbring-at-goodyear.com
- May Contain Confidential and/or Proprietary Information. May not be copied or disseminated without the expressed written consent of The Goodyear Tire & Rubber Company. -
trent-at-ornl.gov
02/08/07 05:08 PM To petra.wahlbring-at-goodyear.com cc Please respond to trent-at-ornl.gov Subject [Microscopy] Cryo ultramicrotomy -help!
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
I have just recently begun using the Leica EM FCS cryo set-up for our ultramicrotome. I'm having a little difficulty and was hoping that members of this list could clue me into some tricks of the trade. I am presently trying to section polymer samples and I need them to be around 50nm in thickness. During cutting, the sections often curl or fold like an accordion. So I pick them up with an eyelash tool and try (with much difficulty and cold fingers) to unfold and stretch them out onto a copper grid. The sections have no affinity for the Cu grid and are much happier adhering to the eyelash or wadding up into an unusable lump. Short sections don't work any better, since these just tend to flip away. I'm using a anti-static device but it's difficult to place it into the correct position since I'm using it's holder to hold the copper grids close to the diamond.
Thank you for any advice! Shawn
==============================Original Headers============================== 3, 30 -- From trent-at-ornl.gov Thu Feb 8 10:04:13 2007 3, 30 -- Received: from emroute3.ornl.gov (emroute3.ornl.gov [160.91.4.110]) 3, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l18G4DcX010021 3, 30 -- for {Microscopy-at-Microscopy.Com} ; Thu, 8 Feb 2007 10:04:13 -0600 3, 30 -- Received: from emroute3.ornl.gov ([127.0.0.1]) 3, 30 -- by emroute3.ornl.gov (PMDF V6.3-x3 #31246) 3, 30 -- with ESMTP id {0JD500FGMJ4X97-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from CONVERSION-DAEMON.emroute3.ornl.gov by emroute3.ornl.gov 3, 30 -- (PMDF V6.3-x3 #31246) id {0JD500G01J4XX8-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from ORNLEXCHANGE.ornl.gov (ornlexchange1.ornl.gov [160.91.1.20]) 3, 30 -- by emroute3.ornl.gov (PMDF V6.3-x3 #31246) 3, 30 -- with ESMTP id {0JD500FBBJ4X9V-at-emroute3.ornl.gov} for 3, 30 -- Microscopy-at-Microscopy.Com; Thu, 08 Feb 2007 11:00:33 -0500 (EST) 3, 30 -- Received: from 160.91.157.36 ([160.91.157.36]) 3, 30 -- by ORNLEXCHANGE.ornl.gov ([160.91.1.32]) via Exchange Front-End Server 3, 30 -- mail.ornl.gov ([160.91.4.23]) with Microsoft Exchange Server HTTP-DAV ; Thu, 3, 30 -- 08 Feb 2007 16:00:10 +0000 3, 30 -- Date: Thu, 08 Feb 2007 11:00:30 -0500 3, 30 -- From: Shawn Reeves {trent-at-ornl.gov} 3, 30 -- Subject: Cryo ultramicrotomy -help! 3, 30 -- To: Microscopy-at-Microscopy.Com 3, 30 -- Message-id: {C1F0B34E.1804%trent-at-ornl.gov} 3, 30 -- MIME-version: 1.0 3, 30 -- Content-type: text/plain; charset=US-ASCII 3, 30 -- Content-transfer-encoding: 7bit 3, 30 -- User-Agent: Microsoft-Entourage/11.3.3.061214 3, 30 -- Thread-Topic: Cryo ultramicrotomy -help! 3, 30 -- Thread-Index: AcdLmkGKgGPTdreNEduLiwAKlebEPA== ==============================End of - Headers==============================
==============================Original Headers============================== 22, 16 -- From petra.wahlbring-at-goodyear.com Fri Feb 9 01:55:52 2007 22, 16 -- Received: from emeam1.goodyear.com (emeam1.goodyear.com [57.67.226.7]) 22, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l197tqYJ014525 22, 16 -- for {Microscopy-at-Microscopy.Com} ; Fri, 9 Feb 2007 01:55:52 -0600 22, 16 -- In-Reply-To: {200702081608.l18G8uot016707-at-ns.microscopy.com} 22, 16 -- Subject: Re: [Microscopy] Cryo ultramicrotomy -help! 22, 16 -- To: trent-at-ornl.gov 22, 16 -- Cc: Microscopy-at-Microscopy.Com 22, 16 -- X-Mailer: Lotus Notes Release 6.5 September 26, 2003 22, 16 -- Message-ID: {OF2115C3C7.24FEE0BC-ONC125727D.002AA858-C125727D.002B5A21-at-goodyear.com} 22, 16 -- From: petra.wahlbring-at-goodyear.com 22, 16 -- Date: Fri, 9 Feb 2007 08:53:30 +0100 22, 16 -- X-MIMETrack: Serialize by Router on ECLNGM01/EU/GDYRNET(Release 6.5.4FP3|January 09, 2006) at 22, 16 -- 02/09/2007 08:55:52 AM 22, 16 -- MIME-Version: 1.0 22, 16 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 34, 22 -- From jfish-at-gladstone.ucsf.edu Fri Feb 9 10:33:11 2007 34, 22 -- Received: from gladstone.ucsf.edu (gladstone.ucsf.edu [169.230.76.25]) 34, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19GXBZk004556 34, 22 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 9 Feb 2007 10:33:11 -0600 34, 22 -- Received: from [169.230.76.4] (HELO JFISH) 34, 22 -- by gladstone.ucsf.edu (CommuniGate Pro SMTP 4.2.10) 34, 22 -- with ESMTP id 563015891 for Microscopy-at-MSA.Microscopy.Com; Fri, 09 Feb 2007 08:33:09 -0800 34, 22 -- Reply-To: {jfish-at-gladstone.ucsf.edu} 34, 22 -- From: "Jo Dee Fish" {jfish-at-gladstone.ucsf.edu} 34, 22 -- To: {Microscopy-at-msa.microscopy.com} 34, 22 -- Subject: RE: [Microscopy] Re: Cryo ultramicrotomy -help! 34, 22 -- Date: Fri, 9 Feb 2007 08:33:05 -0800 34, 22 -- Organization: J. David Gladstone Institutes 34, 22 -- Message-ID: {000301c74c67$f9a28a70$8903010a-at-JFISH} 34, 22 -- MIME-Version: 1.0 34, 22 -- Content-Type: text/plain; 34, 22 -- charset="us-ascii" 34, 22 -- Content-Transfer-Encoding: 7bit 34, 22 -- X-Mailer: Microsoft Office Outlook 11 34, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 34, 22 -- Thread-Index: AcdMIJQO7aXWFq5xSSiMHSwkoDOwNAARdS8g 34, 22 -- In-Reply-To: {200702090801.l1981wF4020908-at-ns.microscopy.com} ==============================End of - Headers==============================
John Mackenzie, Jr. Coordinator for the Center for Electron Microscopy Professor of Microbiology North Carolina State University Phone (919) 515-2664 Fax (919) 515-8293
skperkin-at-vt.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 } ---------------------------------------------------------------------------- } } Hello- } } If Photoshop is used to modify a digital TEM image to remove stain } precipitate, how should the image processing be described in the } figure legend......or is it even ethical to modify an image in this } way? } } Thank you in advance for your opinions! } } }
==============================Original Headers============================== 6, 21 -- From john_mackenzie-at-ncsu.edu Fri Feb 9 10:33:58 2007 6, 21 -- Received: from uni00mr.unity.ncsu.edu (uni00mr.unity.ncsu.edu [152.1.1.163]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19GXuwW005239 6, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 10:33:57 -0600 6, 21 -- Received: from [127.0.0.1] (cord1.mbio.ncsu.edu [152.1.178.41]) 6, 21 -- by uni00mr.unity.ncsu.edu (8.13.7/8.13.8/Nv5.2006.1109) with ESMTP id l19GXqdO018249 6, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 11:33:53 -0500 (EST) 6, 21 -- Message-ID: {45CCA27D.7090808-at-ncsu.edu} 6, 21 -- Date: Fri, 09 Feb 2007 11:34:05 -0500 6, 21 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 6, 21 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) 6, 21 -- MIME-Version: 1.0 6, 21 -- To: microscopy-at-microscopy.com 6, 21 -- Subject: Re: [Microscopy] image processing 6, 21 -- References: {200702091609.l19G9WJj031033-at-ns.microscopy.com} 6, 21 -- In-Reply-To: {200702091609.l19G9WJj031033-at-ns.microscopy.com} 6, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 21 -- Content-Transfer-Encoding: 7bit 6, 21 -- X-PMX-Version: 5.2.1.279297, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.2.9.81933 6, 21 -- X-Spam-Status: No, Hits=7% 6, 21 -- X-Spam-Level: IIIIIII ==============================End of - Headers==============================
The official MSA guidelines require that any change in the image such as this be fully documented.
My opinion: I would doubt that this operation would be ethical as it fundamentally changes your original data. I doubt that the image would pass review once the altering of the image was disclosed. What is preventing you from resectioning and staining properly?
John
John Mackenzie, Jr. Coordinator for the Center for Electron Microscopy Professor of Microbiology North Carolina State University Phone (919) 515-2664 Fax (919) 515-8293
skperkin-at-vt.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 } ---------------------------------------------------------------------------- } } Hello- } } If Photoshop is used to modify a digital TEM image to remove stain } precipitate, how should the image processing be described in the } figure legend......or is it even ethical to modify an image in this } way? } } Thank you in advance for your opinions! } } }
==============================Original Headers============================== 8, 21 -- From john_mackenzie-at-ncsu.edu Fri Feb 9 10:38:25 2007 8, 21 -- Received: from uni00mr.unity.ncsu.edu (uni00mr.unity.ncsu.edu [152.1.1.163]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19GcPHt015903 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 10:38:25 -0600 8, 21 -- Received: from [127.0.0.1] (cord1.mbio.ncsu.edu [152.1.178.41]) 8, 21 -- by uni00mr.unity.ncsu.edu (8.13.7/8.13.8/Nv5.2006.1109) with ESMTP id l19GcNaA020864 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 11:38:24 -0500 (EST) 8, 21 -- Message-ID: {45CCA38A.2000508-at-ncsu.edu} 8, 21 -- Date: Fri, 09 Feb 2007 11:38:34 -0500 8, 21 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 8, 21 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) 8, 21 -- MIME-Version: 1.0 8, 21 -- To: microscopy-at-microscopy.com 8, 21 -- Subject: Re: [Microscopy] image processing 8, 21 -- References: {200702091609.l19G9WJj031033-at-ns.microscopy.com} 8, 21 -- In-Reply-To: {200702091609.l19G9WJj031033-at-ns.microscopy.com} 8, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- X-PMX-Version: 5.2.1.279297, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.2.9.82933 8, 21 -- X-Spam-Status: No, Hits=7% 8, 21 -- X-Spam-Level: IIIIIII ==============================End of - Headers==============================
I don't think you should really remove any artifacts or background - look at JCB vol. 166, p. 11 (2004) for a pretty good editorial.
What they consider OK is just level adjustment. Even if you are touching the curves, you should state it and removing/adding anything is not acceptable at all.
Michal
skperkin-at-vt.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 } ---------------------------------------------------------------------------- } } Hello- } } If Photoshop is used to modify a digital TEM image to remove stain } precipitate, how should the image processing be described in the } figure legend......or is it even ethical to modify an image in this } way? } } Thank you in advance for your opinions! } } }
==============================Original Headers============================== 5, 19 -- From Michal.Jarnik-at-fccc.edu Fri Feb 9 10:40:03 2007 5, 19 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19Ge3ZB021565 5, 19 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 10:40:03 -0600 5, 19 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) 5, 19 -- (authenticated bits=0) 5, 19 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id l19Ge2Yh017838 5, 19 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 11:40:02 -0500 (EST) 5, 19 -- Message-ID: {45CCA3E0.604-at-fccc.edu} 5, 19 -- Date: Fri, 09 Feb 2007 11:40:00 -0500 5, 19 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} 5, 19 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: microscopy-at-microscopy.com 5, 19 -- Subject: Re: [Microscopy] image processing 5, 19 -- References: {200702091610.l19GALNn001549-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200702091610.l19GALNn001549-at-ns.microscopy.com} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Years ago, I had a student develop a program that would convert images from Oxford's ISIS Labbook/Job storage system to TIFF format in bulk. I suppose we could have done something with the spectra at the same time, but we didn't.
For as many spectra as we collect, I cannot imagine converting them all to another format. What would you do with them all? Instead, I wrote a rather simple macro to aid in placing selected spectra into Word documents and use that when necessary.
But to answer your question, David Vowles at Cambridge (djv21-at-cam.ac.uk) wrote a package called Spectrum Plot which reads and displays multiple formats of files and can export them into multiple formats. It also has a batch conversion option.
I have not heard from David in a while, but I should still have a copy of the installation files in case you cannot find the elsewhere.
Warren Straszheim
-----Original Message----- X-from: jehrman-at-mta.ca [mailto:jehrman-at-mta.ca] Sent: Friday, February 09, 2007 9:11 AM To: wesaia-at-iastate.edu
OK, I've had enough. Has anybody come up with a way to export all the spectra in an Oxford Inca Project (to EMSA format in particular) in one fell swoop? I'm getting a little tired the right click fandango for each and every spectra. If it matters, I have Inca 200 version 4.02.
My worst nightmare: Sure, version x.xx can do that. It's only a zillion dollars. I don't need any more pretty graphics and hand holding. I'd just like to get mundane grunt work done fast. After all, that's what computers are for.....
At least it's Friday.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have extended experience in polymer samples and here are some suggestions: -use a wire loop attached to a wood BBQ stick to collect the sections from the cryo-chamber. The wire loop could be about 3mm in diameter. -dip the loop in 1-2%sucrose solution and insert it into the chamber. -wait until you see the solution in the loop start freezing and then aproach the loop to the section to collect them. -remove the loop from the chamber and allow the solution to un-freeze. -while looking through a stereo scope, place the section on the grid which is holded with tweezers. -remove the excess solution from the grid with a small piece of filter paper through the place between the tweezer's fingers. -place the grid on plenty diH2O in a beaker with the section facing the water until you are done sectioning, to remove the sucrose solution. -remove the grids and dry them as mentioned above.
I hope this helps. If you have further questions, email me or just call.
ANI M ISSAIAN Electron Microscopy Facility Manager California State University, Northridge 18111 Nordhoff street, Northridge, CA 91330-8303 Biology dept., MC 8303, Room CS2205 Phone: (818) 677-3383 Fax (818) 677-2034
---- Original message ---- } Date: Thu, 8 Feb 2007 10:11:35 -0600 } From: trent-at-ornl.gov } Subject: [Microscopy] Cryo ultramicrotomy -help! } To: ani.issaian-at-csun.edu } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 8, 28 -- From ani.issaian-at-csun.edu Fri Feb 9 11:26:25 2007 8, 28 -- Received: from plover.csun.edu (plover.csun.edu [130.166.1.24]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19HQOvl028852 8, 28 -- for {Microscopy-at-Microscopy.Com} ; Fri, 9 Feb 2007 11:26:25 -0600 8, 28 -- Received: from puffin.csun.edu (puffin.csun.edu [130.166.1.21]) 8, 28 -- by plover.csun.edu (MOS 3.8.2-GA) 8, 28 -- with ESMTP id DND24083; 8, 28 -- Fri, 9 Feb 2007 09:26:20 -0800 (PST) 8, 28 -- Received: from cuckoo.csun.edu (cuckoo.csun.edu [130.166.1.230]) 8, 28 -- by puffin.csun.edu (MOS 3.7.5a-GA) 8, 28 -- with ESMTP id FKW05722; 8, 28 -- Fri, 9 Feb 2007 09:26:19 -0800 (PST) 8, 28 -- Received: (from cuckoo.csun.edu [130.166.114.202]) 8, 28 -- by cuckoo.csun.edu (MOS 3.8.2-GA) 8, 28 -- with HTTPS/1.1 id ATE50444 (AUTH ami24015); 8, 28 -- Fri, 9 Feb 2007 09:26:18 -0800 (PST) 8, 28 -- From: Ani M Issaian {ani.issaian-at-csun.edu} 8, 28 -- Subject: Re: [Microscopy] Cryo ultramicrotomy -help! 8, 28 -- To: trent-at-ornl.gov 8, 28 -- Cc: Microscopy-at-Microscopy.Com 8, 28 -- Reply-To: ani.issaian-at-csun.edu 8, 28 -- X-Mailer: Mirapoint Webmail Direct 3.8.2-GA 8, 28 -- MIME-Version: 1.0 8, 28 -- Content-Type: text/plain; charset=us-ascii 8, 28 -- Content-Transfer-Encoding: 7bit 8, 28 -- Message-Id: {20070209092618.ATE50444-at-cuckoo.csun.edu} 8, 28 -- Date: Fri, 9 Feb 2007 09:26:18 -0800 (PST) 8, 28 -- X-Junkmail-Whitelist: YES (by domain whitelist at plover.csun.edu) ==============================End of - Headers==============================
} We have developed a new three-step process to clean moly apertures } so that all extraneous material, including flashing, is removed } during our aperture manufacturing. Please contact us directly and } we'll see if it's possible to clean up your used apertures. } } John Arnott
Disclaimer: Ladd Research produces Apertures and Microholes for EM use and other custom work
} Ladd Research } 83 Holly Court } Williston, VT 05945 } } On-line Catalog: www.laddresearch.com } } Telephone: 1-802-658-4961 (anywhere) } Toll Free 1-800-451-3406 (US) } Fax: 1-802-660-8859 } } e-mail: sales-at-laddresearch.com } } Disclaimer: Ladd Research is a manufacturer of apertures and other } electron microscopy products } } } At 04:13 PM 2/7/2007, you wrote: } ---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Does anyone have any experience cleaning the carbonaeous gunk off of Mo or Pt-Ir apertures with a solvent? I know about heating up in a Mo etc boat, but looking for alternatives. Will ammonium hydroxide do the trick? Or something else?
thanks. -- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
Thanks to all who have replied about my grumblings. But I think I need to clarify. I don't have any problem actually *performing* the export from Inca to EMSA. The Inca software does this just fine. The problem is when I have *50* of these things to convert to EMSA. I can't find any way to do it in the INCA interface other than right-clicking, choosing export, choosing EMSA, getting the file dialog, possibly modifying the name of the file, and hitting OK. The *&(#*&!!!! interface *will* let you select multiple spectra from the data pane, but the only option available on the right-click context menu is Delete! It appears that Dilbert's company had something to do with the design here....
As to why I want to do this for so many spectra - I have a project where I need to pull off specific values from certain channels in the spectra. I've written a program to do that (in batch, takes all of 2 seconds for 50 spectra), once I have the spectra in EMSA format. It's the mind-numbing, senseless waste of time involved in clicking through all those spectra to get the EMSA version in the first place that's driving me nuts. For those who aren't familiar with the Inca software, all the spectra are wrapped up in an "ipj" extension file. Those spectra can be exported one at a time to almost any format, but one can't do it en masse.
Oh yeah, I also export spectra to EMSA when I need a publication quality figure. The figures generated by the Reporting module for the Inca system just don't cut it.
I have thought of a way to do this, using a macro utility like Macro Express, but things like this haven't run terribly reliably for me on the old NT 4.0 system of the INCA.
Maybe someone from Oxford is listening? Perusing the gazillions of EXE and DLL files that make up this package, possibly one of them contains the export function that can be called from another program. I'll hold my breath while I wait for a reply.....
Still, it is Friday....
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Method: "Image was artistically rendered to resemble what we expect it would look like (a.k.a. to support our argument) were the staining procedure optimized."
} The official MSA guidelines require that any change in the image such as } this be fully documented. } } My opinion: I would doubt that this operation would be ethical as it } fundamentally changes your original data. I doubt that the image would } pass review once the altering of the image was disclosed. What is } preventing you from resectioning and staining properly? } } John } } John Mackenzie, Jr. } Coordinator for the Center for Electron Microscopy } Professor of Microbiology } North Carolina State University } Phone (919) 515-2664 Fax (919) 515-8293 } } } } skperkin-at-vt.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 } } } ---------------------------------------------------------------------------- } } } } Hello- } } } } If Photoshop is used to modify a digital TEM image to remove stain } } precipitate, how should the image processing be described in the } } figure legend......or is it even ethical to modify an image in this } } way? } } } } Thank you in advance for your opinions! } } } } } } } } } ==============================Original Headers============================== } 8, 21 -- From john_mackenzie-at-ncsu.edu Fri Feb 9 10:38:25 2007 } 8, 21 -- Received: from uni00mr.unity.ncsu.edu } (uni00mr.unity.ncsu.edu [152.1.1.163]) } 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l19GcPHt015903 } 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 } 10:38:25 -0600 } 8, 21 -- Received: from [127.0.0.1] (cord1.mbio.ncsu.edu [152.1.178.41]) } 8, 21 -- by uni00mr.unity.ncsu.edu } (8.13.7/8.13.8/Nv5.2006.1109) with ESMTP id l19GcNaA020864 } 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 } 11:38:24 -0500 (EST) } 8, 21 -- Message-ID: {45CCA38A.2000508-at-ncsu.edu} } 8, 21 -- Date: Fri, 09 Feb 2007 11:38:34 -0500 } 8, 21 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} } 8, 21 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) } 8, 21 -- MIME-Version: 1.0 } 8, 21 -- To: microscopy-at-microscopy.com } 8, 21 -- Subject: Re: [Microscopy] image processing } 8, 21 -- References: {200702091609.l19G9WJj031033-at-ns.microscopy.com} } 8, 21 -- In-Reply-To: {200702091609.l19G9WJj031033-at-ns.microscopy.com} } 8, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 8, 21 -- Content-Transfer-Encoding: 7bit } 8, 21 -- X-PMX-Version: 5.2.1.279297, Antispam-Engine: 2.5.0.283055, } Antispam-Data: 2007.2.9.82933 } 8, 21 -- X-Spam-Status: No, Hits=7% } 8, 21 -- X-Spam-Level: IIIIIII } ==============================End of - Headers==============================
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 8, 30 -- From cammer-at-aecom.yu.edu Fri Feb 9 13:35:47 2007 8, 30 -- Received: from mx1.aecom.yu.edu (mx1.aecom.yu.edu [129.98.1.51]) 8, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19JZkDq005780 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 13:35:47 -0600 8, 30 -- Received: from draco.aecom.yu.edu (draco.aecom.yu.edu [129.98.1.160]) 8, 30 -- by mx1.aecom.yu.edu (Postfix) with ESMTP id B24609F0030 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:35:46 -0500 (EST) 8, 30 -- Received: from draco.aecom.yu.edu (unknown [127.0.0.1]) 8, 30 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 40CE18B4009 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:22:57 -0500 (EST) 8, 30 -- X-AuditID: 816201a0-a16c9bb0000017bb-24-45ccca11904f 8, 30 -- Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.100]) 8, 30 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 0BF25718002 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:22:57 -0500 (EST) 8, 30 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.80.70]) 8, 30 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 8, 30 -- (No client certificate requested) 8, 30 -- by post.aecom.yu.edu (Postfix) with ESMTP id 3ADFB28 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:35:46 -0500 (EST) 8, 30 -- Message-Id: {7.0.1.0.2.20070209143238.054c8178-at-aecom.yu.edu} 8, 30 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 8, 30 -- Date: Fri, 09 Feb 2007 14:33:43 -0500 8, 30 -- To: microscopy-at-microscopy.com 8, 30 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 8, 30 -- Subject: Re: [Microscopy] Re: image processing 8, 30 -- In-Reply-To: {200702091639.l19Gdw6J021233-at-ns.microscopy.com} 8, 30 -- References: {200702091639.l19Gdw6J021233-at-ns.microscopy.com} 8, 30 -- Mime-Version: 1.0 8, 30 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 30 -- X-Brightmail-Tracker: AAAAAA== ==============================End of - Headers==============================
We want to try embedding zebrafish in polyester wax/carbowax. Any thoughts, suggestions, protocols, references, vendors, etc. would be welcome. Thanks!
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 6, 33 -- From mcauliff-at-umdnj.edu Fri Feb 9 15:43:23 2007 6, 33 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19LhMjh019617 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Fri, 9 Feb 2007 15:43:23 -0600 6, 33 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 6, 33 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 4092F4BED0 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Fri, 9 Feb 2007 16:43:22 -0500 (EST) 6, 33 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 6, 33 -- by zix02.umdnj.edu (Proprietary) with ESMTP id 66CCA4BE81 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Fri, 9 Feb 2007 16:43:21 -0500 (EST) 6, 33 -- Received: from ([130.219.34.131]) 6, 33 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.75390795; 6, 33 -- Fri, 09 Feb 2007 16:42:57 -0500 6, 33 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 33 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 33 -- id {0JD700001T88XY-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 33 -- for microscopy-at-msa.microscopy.com; Fri, 09 Feb 2007 16:42:57 -0500 (EST) 6, 33 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 33 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 33 -- 2004)) with ESMTP id {0JD700F65TNE8Z-at-Polaris.umdnj.edu} ; Fri, 6, 33 -- 09 Feb 2007 16:42:51 -0500 (EST) 6, 33 -- Date: Fri, 09 Feb 2007 16:44:15 -0500 6, 33 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 33 -- Subject: carbowax/polyester wax 6, 33 -- To: Histonet {histonet-at-pathology.swmed.edu} , 6, 33 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 33 -- Message-id: {45CCEB2F.6030901-at-umdnj.edu} 6, 33 -- MIME-version: 1.0 6, 33 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 33 -- Content-transfer-encoding: 7BIT 6, 33 -- X-Accept-Language: en-us, en 6, 33 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 33 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
didn't see a reply to this - I suspect it is there for osmotic purposes. I believe LR Gold embedding protocols sometimes include PVP with the ethanol dehydration steps to minimize osmotic stress.
At 12:01 PM 02/07/07, you wrote:
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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
I'm looking for the most efficient and accurate way to trace features in digital images (for instance, outlining different minerals or metal phases in backscattered electron images). Does anyone have experience with this? There seems to be three basic ways to go: (1) graphics pads that replace a mouse, (2) Tablet PCs, and (3) "pen displays" -- that is, LCD displays on which, like Tablet PCs, a stylus can write on the screen. Any relevant experiences out there with such devices? Any insights would be welcome. Feel free to email me off-listserver with any comments or suggestions. If there is a demand, I can post a summary of the answers.
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu Personal Website: http://umn.edu/~frah0010
==============================Original Headers============================== 8, 20 -- From frah0010-at-umn.edu Fri Feb 9 20:44:31 2007 8, 20 -- Received: from mta-m2.tc.umn.edu (mta-m2.tc.umn.edu [160.94.23.21]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1A2iUKV014238 8, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 20:44:31 -0600 8, 20 -- Received: from [10.0.1.3] (c-75-72-182-252.hsd1.mn.comcast.net [75.72.182.252]) 8, 20 -- by mta-m2.tc.umn.edu (UMN smtpd) with ESMTP 8, 20 -- Fri, 9 Feb 2007 20:44:29 -0600 (CST) 8, 20 -- X-Umn-Remote-Mta: [N] c-75-72-182-252.hsd1.mn.comcast.net [75.72.182.252] #+TS+AU+HN 8, 20 -- In-Reply-To: {1D190879-3592-4F6F-B0BE-857EB4840C8A-at-umn.edu} 8, 20 -- References: {1D190879-3592-4F6F-B0BE-857EB4840C8A-at-umn.edu} 8, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 20 -- Message-Id: {A3D8A22D-6E52-41EA-925F-6FACADA2DB4C-at-umn.edu} 8, 20 -- Cc: Ellery Frahm {frah0010-at-umn.edu} 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- From: Ellery Frahm {frah0010-at-umn.edu} 8, 20 -- Subject: digital sketching 8, 20 -- Date: Fri, 9 Feb 2007 20:44:22 -0600 8, 20 -- To: Microscopy-at-microscopy.com 8, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
The website for the M&M 2007 meeting says that the submission deadline is Wed Feb 15, 2007. Feb 15 is a Thursday on my calendar. Can anybody set me straight? I can't find my call for papers.
Thanks in advance
-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 Headers============================== 4, 20 -- From walck-at-southbaytech.com Fri Feb 9 20:55:07 2007 4, 20 -- Received: from flpi102.sbcis.sbc.com (flpi102.sbcis.sbc.com [207.115.20.71]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1A2t78m025426 4, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 20:55:07 -0600 4, 20 -- X-ORBL: [64.169.217.123] 4, 20 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 4, 20 -- by flpi102.sbcis.sbc.com (8.13.8 out.dk.spool/8.13.8) with ESMTP id l1A2t5JY025273 4, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 18:55:06 -0800 4, 20 -- From: "Scott Walck" {walck-at-southbaytech.com} 4, 20 -- To: {Microscopy-at-microscopy.com} 4, 20 -- Subject: Submission deadline 4, 20 -- Date: Fri, 9 Feb 2007 18:55:15 -0800 4, 20 -- Message-ID: {005a01c74cbe$e4ba99c0$7801a8c0-at-dynamicbl8uno3} 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: text/plain; 4, 20 -- charset="us-ascii" 4, 20 -- Content-Transfer-Encoding: 7bit 4, 20 -- X-Mailer: Microsoft Office Outlook 11 4, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 4, 20 -- Thread-Index: AcdMvuQfw00QSO25QAWK7gwDn7jIfg== ==============================End of - Headers==============================
When producing Vibratome sections (for EM or light microscopy) of insect tissue, I usually embed the tissue in 15% gelatine before sectioning, and have been doing that for years. But now I suddenly have grown tired of the sticky gelatinous mass that surrounds these fragile sections. Agarose is supposed to be a good substitute- has anyone got experience with it or can tell me which type/temperature/concentration of agarose to test?
thank you
Gerd Leitinger
Dr Gerd Leitinger Institute of Cell Biology, Histology, and Embryology Center of Molecular Medicine Medical University of Graz Harrachgasse 21 8010 Graz Austria
phone +43 316 380 4237
==============================Original Headers============================== 6, 18 -- From Gerd.Leitinger-at-meduni-graz.at Sat Feb 10 02:31:33 2007 6, 18 -- Received: from viefep15-int.chello.at (viefep18-int.chello.at [213.46.255.21]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1A8VWd7009097 6, 18 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 02:31:32 -0600 6, 18 -- Received: from [84.115.150.93] by viefep15-int.chello.at 6, 18 -- (InterMail vM.6.01.05.04 201-2131-123-105-20051025) with ESMTP 6, 18 -- id {20070210083130.VLZ21314.viefep15-int.chello.at-at-[84.115.150.93]} 6, 18 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 09:31:30 +0100 6, 18 -- Message-ID: {45CD82E0.8020407-at-meduni-graz.at} 6, 18 -- Date: Sat, 10 Feb 2007 09:31:28 +0100 6, 18 -- From: "Dr. Gerd Leitinger" {Gerd.Leitinger-at-meduni-graz.at} 6, 18 -- Reply-To: Gerd.Leitinger-at-meduni-graz.at 6, 18 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 6, 18 -- MIME-Version: 1.0 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- Subject: Vibratome sections 6, 18 -- Content-Type: text/plain; charset=ISO-8859-15; format=flowed 6, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I originally found using a graphics tablet (a cheap £70 Wacon Graphire 2 pad) a real pain to get used to and stuck with a mouse. I did have lot of success with a light pen years ago (c1990) using a £80k Magiscan Colour image analyser, but that pen was integrated into the software/hardware/VDU. Light pens are still available, but I never bothered investigating them as they are relatively expensive.
However when I had to trace around hundreds of cells that couldn't be detected by thresholding, I found a mouse just wasn't accurate enough (I did think of trying a far higher dpi gaming mouse). I was using MetaMorph image anaylsis software. MetaMorph's main failing is no binary editor to remove/add incorrectly thresholded regions so manual thesholding simply couldn't be used to detect the cells.
Drawing round manually with the mouse was such a pain that I got the Graphire 2 out of the drawer and plugged it in (it's USB so it was easy when it's drivers are loaded, and the USB MS mouse still works). I found that within a few minutes I was drawing round the cells far more easily, and now I always go for the Graphire 2 pad for such things. A lot of the ease depends on the quality of the program you are using the pad with though, as MetaMorph and Image Pro Plus offer 'back untrace' undo options if you slip off the edge.
I have a more recent even cheaper Graphire pad 4 pad (£40), and I have to say the position of the click buttons is very poorly located on the new pen (for me) compared to my old Graphire 2 tablet (£70). You could try the upmarket Intuos (£200) that offer A4+ sizes (and with all the Wacom tablets you can put photos under a pad 'flap' to trace over). Wacom also provide a crosshair optical viewfinder mouse for accurate tracing over photos using the Intuos, just like expensive CAD tablets (see wacom.com). I didn't like the cheap Wacom mouse that came with the Graphire 2 pad though - it's too unresponsive - no doubt the expensive Intuos one is a lot better. For home use I got a student copy of Corel painter IX for the kids, as this works great with a Wacom tablet (press harder on the tablet with the pen and the line gets thicker, like a real paintbrush) - they soon lost my stylus pen though (£28).
Always try an image analysis program (e.g. ImageJ / NIHImage which is free), to see if you can manually threshold the areas you want to detect, for very quick and easy measurement. It's always worth taking time with specimen preparation and image capture to create pictures that are highly conducive to this.
I have always found laptops and tablet PCs too slow compared to an imaging workstation, although the new Intel Duo 2 has improved their performance somewhat - but there's still few upgrade paths when the hardware becomes dated compared to a standard PC, and they are more expensive to buy (particularly if, like me, you want one able to play Doom 3 occasionally).
So tracing objects like cells with cheap Wacom Graphire tablet and stylus pen worked for me. You still need the PC's mouse for menu's etc as the pen is too slow for this (they both work together happily).
Regards
Keith
-------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu] Sent: 10 February 2007 02:49 To: keith.morris-at-ucl.ac.uk
Microscopy folks,
I'm looking for the most efficient and accurate way to trace features in digital images (for instance, outlining different minerals or metal phases in backscattered electron images). Does anyone have experience with this? There seems to be three basic ways to go: (1) graphics pads that replace a mouse, (2) Tablet PCs, and (3) "pen displays" -- that is, LCD displays on which, like Tablet PCs, a stylus can write on the screen. Any relevant experiences out there with such devices? Any insights would be welcome. Feel free to email me off-listserver with any comments or suggestions. If there is a demand, I can post a summary of the answers.
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu Personal Website: http://umn.edu/~frah0010
==============================Original Headers============================== 8, 20 -- From frah0010-at-umn.edu Fri Feb 9 20:44:31 2007 8, 20 -- Received: from mta-m2.tc.umn.edu (mta-m2.tc.umn.edu [160.94.23.21]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1A2iUKV014238 8, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 20:44:31 -0600 8, 20 -- Received: from [10.0.1.3] (c-75-72-182-252.hsd1.mn.comcast.net [75.72.182.252]) 8, 20 -- by mta-m2.tc.umn.edu (UMN smtpd) with ESMTP 8, 20 -- Fri, 9 Feb 2007 20:44:29 -0600 (CST) 8, 20 -- X-Umn-Remote-Mta: [N] c-75-72-182-252.hsd1.mn.comcast.net [75.72.182.252] #+TS+AU+HN 8, 20 -- In-Reply-To: {1D190879-3592-4F6F-B0BE-857EB4840C8A-at-umn.edu} 8, 20 -- References: {1D190879-3592-4F6F-B0BE-857EB4840C8A-at-umn.edu} 8, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 20 -- Message-Id: {A3D8A22D-6E52-41EA-925F-6FACADA2DB4C-at-umn.edu} 8, 20 -- Cc: Ellery Frahm {frah0010-at-umn.edu} 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- From: Ellery Frahm {frah0010-at-umn.edu} 8, 20 -- Subject: digital sketching 8, 20 -- Date: Fri, 9 Feb 2007 20:44:22 -0600 8, 20 -- To: Microscopy-at-microscopy.com 8, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
==============================Original Headers============================== 31, 24 -- From keith.morris-at-ucl.ac.uk Sat Feb 10 04:43:26 2007 31, 24 -- Received: from smtp2.global.net.uk (smtp2.global.net.uk [80.189.94.52]) 31, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1AAhQvd022887 31, 24 -- for {Microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 04:43:26 -0600 31, 24 -- Received: from 100.237.adsl.brightview.com ([80.189.237.100] helo=loungepc) 31, 24 -- by smtp2.global.net.uk with esmtp (Exim 4.42) 31, 24 -- id 1HFphT-000Doz-3S; Sat, 10 Feb 2007 10:43:25 +0000 31, 24 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 31, 24 -- To: {frah0010-at-umn.edu} 31, 24 -- Cc: {Microscopy-at-microscopy.com} 31, 24 -- References: {200702100249.l1A2nTxm020866-at-ns.microscopy.com} 31, 24 -- Subject: RE: [Microscopy] digital sketching 31, 24 -- Date: Sat, 10 Feb 2007 10:43:21 -0000 31, 24 -- Message-ID: {000001c74d00$4ab4cc50$0301a8c0-at-loungepc} 31, 24 -- MIME-Version: 1.0 31, 24 -- Content-Type: text/plain; 31, 24 -- charset="iso-8859-1" 31, 24 -- X-Mailer: Microsoft Office Outlook 11 31, 24 -- Thread-Index: AcdMvhteFNQjJDLOS+6R0XsAno+y1QAOrXkg 31, 24 -- In-Reply-To: {200702100249.l1A2nTxm020866-at-ns.microscopy.com} 31, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 31, 24 -- Authenticated-Sender: 31, 24 -- Content-Transfer-Encoding: 8bit 31, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1AAhQvd022887 ==============================End of - Headers==============================
It's image editing really not image processing. I would have thought that if you showed the original and the 'doctored' version together all should be fine. However journal peer review is so 'complicated' at present, even this may not be acceptable to many (these days you seem to have to tell a complete polished story rather than just present new data for discussion).
When you push a button on a digital camera the resulting image has been subjected to intense image processing (sharpen, light balance, noise reduction) already (and you can get the angle right, crop or introduce shadows to remove any unpleasantness). Deconvolution software also applies a lot of processing to the images. Plus using a confocal microscope and a mixture of flourescent labels it's a small matter of adjusting PMT gain etc.. to significantly transform the picture to more like that of an 'image rendered to resemble what we expect it should look like (a.k.a. to support our argument)' before you click the scan button. This can completely remove less bright staining regions, although the image hasn't technically been doctored. Plus of course you can move to the one area on the specimen where it really shows what you want to see (the rest of the sample being rejected as being a bit too typical). Most images for publication are chosen on aesthetics as much as on scientific grounds, and so are likely to be atypical. No doubt the researcher has used random field selection etc.., to ensure that the image isn't actually misleading though.
Keith
-------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: cammer-at-aecom.yu.edu [mailto:cammer-at-aecom.yu.edu] Sent: 09 February 2007 19:41 To: keith.morris-at-ucl.ac.uk
Method: "Image was artistically rendered to resemble what we expect it would look like (a.k.a. to support our argument) were the staining procedure optimized."
} The official MSA guidelines require that any change in the image such as } this be fully documented. } } My opinion: I would doubt that this operation would be ethical as it } fundamentally changes your original data. I doubt that the image would } pass review once the altering of the image was disclosed. What is } preventing you from resectioning and staining properly? } } John } } John Mackenzie, Jr. } Coordinator for the Center for Electron Microscopy } Professor of Microbiology } North Carolina State University } Phone (919) 515-2664 Fax (919) 515-8293 } } } } skperkin-at-vt.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 } } } ---------------------------------------------------------------------------- } } } } Hello- } } } } If Photoshop is used to modify a digital TEM image to remove stain } } precipitate, how should the image processing be described in the } } figure legend......or is it even ethical to modify an image in this } } way? } } } } Thank you in advance for your opinions! } } } } } } } } } ==============================Original Headers============================== } 8, 21 -- From john_mackenzie-at-ncsu.edu Fri Feb 9 10:38:25 2007 } 8, 21 -- Received: from uni00mr.unity.ncsu.edu } (uni00mr.unity.ncsu.edu [152.1.1.163]) } 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l19GcPHt015903 } 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 } 10:38:25 -0600 } 8, 21 -- Received: from [127.0.0.1] (cord1.mbio.ncsu.edu [152.1.178.41]) } 8, 21 -- by uni00mr.unity.ncsu.edu } (8.13.7/8.13.8/Nv5.2006.1109) with ESMTP id l19GcNaA020864 } 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 } 11:38:24 -0500 (EST) } 8, 21 -- Message-ID: {45CCA38A.2000508-at-ncsu.edu} } 8, 21 -- Date: Fri, 09 Feb 2007 11:38:34 -0500 } 8, 21 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} } 8, 21 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) } 8, 21 -- MIME-Version: 1.0 } 8, 21 -- To: microscopy-at-microscopy.com } 8, 21 -- Subject: Re: [Microscopy] image processing } 8, 21 -- References: {200702091609.l19G9WJj031033-at-ns.microscopy.com} } 8, 21 -- In-Reply-To: {200702091609.l19G9WJj031033-at-ns.microscopy.com} } 8, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 8, 21 -- Content-Transfer-Encoding: 7bit } 8, 21 -- X-PMX-Version: 5.2.1.279297, Antispam-Engine: 2.5.0.283055, } Antispam-Data: 2007.2.9.82933 } 8, 21 -- X-Spam-Status: No, Hits=7% } 8, 21 -- X-Spam-Level: IIIIIII } ==============================End of - Headers==============================
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 8, 30 -- From cammer-at-aecom.yu.edu Fri Feb 9 13:35:47 2007 8, 30 -- Received: from mx1.aecom.yu.edu (mx1.aecom.yu.edu [129.98.1.51]) 8, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l19JZkDq005780 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 13:35:47 -0600 8, 30 -- Received: from draco.aecom.yu.edu (draco.aecom.yu.edu [129.98.1.160]) 8, 30 -- by mx1.aecom.yu.edu (Postfix) with ESMTP id B24609F0030 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:35:46 -0500 (EST) 8, 30 -- Received: from draco.aecom.yu.edu (unknown [127.0.0.1]) 8, 30 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 40CE18B4009 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:22:57 -0500 (EST) 8, 30 -- X-AuditID: 816201a0-a16c9bb0000017bb-24-45ccca11904f 8, 30 -- Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.100]) 8, 30 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 0BF25718002 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:22:57 -0500 (EST) 8, 30 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.80.70]) 8, 30 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 8, 30 -- (No client certificate requested) 8, 30 -- by post.aecom.yu.edu (Postfix) with ESMTP id 3ADFB28 8, 30 -- for {microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 14:35:46 -0500 (EST) 8, 30 -- Message-Id: {7.0.1.0.2.20070209143238.054c8178-at-aecom.yu.edu} 8, 30 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 8, 30 -- Date: Fri, 09 Feb 2007 14:33:43 -0500 8, 30 -- To: microscopy-at-microscopy.com 8, 30 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 8, 30 -- Subject: Re: [Microscopy] Re: image processing 8, 30 -- In-Reply-To: {200702091639.l19Gdw6J021233-at-ns.microscopy.com} 8, 30 -- References: {200702091639.l19Gdw6J021233-at-ns.microscopy.com} 8, 30 -- Mime-Version: 1.0 8, 30 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 30 -- X-Brightmail-Tracker: AAAAAA== ==============================End of - Headers==============================
==============================Original Headers============================== 19, 23 -- From keith.morris-at-ucl.ac.uk Sat Feb 10 05:10:38 2007 19, 23 -- Received: from smtp4.global.net.uk (smtp4.global.net.uk [80.189.92.92]) 19, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1ABAcR8004913 19, 23 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 05:10:38 -0600 19, 23 -- Received: from 11.235.adsl.brightview.com ([80.189.235.11] helo=loungepc) 19, 23 -- by smtp4.global.net.uk with esmtp (Exim 4.42) 19, 23 -- id 1HFq7l-000DyI-Sh 19, 23 -- for microscopy-at-microscopy.com; Sat, 10 Feb 2007 11:10:37 +0000 19, 23 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 19, 23 -- To: {microscopy-at-microscopy.com} 19, 23 -- References: {200702091941.l19Jf8J1012530-at-ns.microscopy.com} 19, 23 -- Subject: RE: [Microscopy] image processing 19, 23 -- Date: Sat, 10 Feb 2007 11:10:28 -0000 19, 23 -- Message-ID: {000001c74d04$16871d30$0301a8c0-at-loungepc} 19, 23 -- MIME-Version: 1.0 19, 23 -- Content-Type: text/plain; 19, 23 -- charset="us-ascii" 19, 23 -- Content-Transfer-Encoding: 7bit 19, 23 -- X-Mailer: Microsoft Office Outlook 11 19, 23 -- Thread-Index: AcdMgkpWXjJFrydoRC6NrFTs2++wUgAdFmAg 19, 23 -- In-Reply-To: {200702091941.l19Jf8J1012530-at-ns.microscopy.com} 19, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 19, 23 -- Authenticated-Sender: ==============================End of - Headers==============================
Here is one of those topics I thought I understood but now that I am teaching it I find myself confused. How can an S wave pass through a phase specimen and NOT interact with it?
I'm using Douglas Murphy's excellent text on light microscopy and digital imaging. In introducing PC optics he states (pg 99):
"Upon transit through a phase object, and incident wave of an illuminating beam becomes divided into two components: (1) an undeviated (0th order) waved or surround wave (S wave) that passes through the specimen but does not interact with it, and (2) a deviated or diffracted wave (D wave) that becomes scattered in many directions." .
At the (also excellent) Molecular Expression web site, Murphy and colleagues say it slightly differently:
"The primary component is an undeviated (or undiffracted; zeroth- order) planar wavefront, commonly referred to as the surround (S) wave, which passes through and around the specimen, but does not interact with it."
How is it that a light wave can pass through a phase object yet not interact with the object and not have its phase changed, while the diffracted light does have its phase changed?
Gary Radice
==============================Original Headers============================== 15, 21 -- From gradice-at-richmond.edu Sun Feb 11 14:35:27 2007 15, 21 -- Received: from nylon.richmond.edu (nylon.richmond.edu [141.166.30.20]) 15, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1BKZQxO014910 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 14:35:26 -0600 15, 21 -- Received: from polyester.richmond.edu (polyester.richmond.edu [141.166.24.28]) 15, 21 -- by nylon.richmond.edu (8.13.1/8.13.1) with ESMTP id l1BKZQ2Q009375 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 15:35:26 -0500 15, 21 -- Received: from [10.0.1.3] (lvs080049.richmond.edu [141.166.80.49]) 15, 21 -- by polyester.richmond.edu (8.12.11.20060308/8.12.11) with ESMTP id l1BKZPVs032335 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 15:35:26 -0500 15, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 15, 21 -- Content-Transfer-Encoding: 7bit 15, 21 -- Message-Id: {79E3462E-2520-4BB2-BAE0-209724B7F43C-at-richmond.edu} 15, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 15, 21 -- To: Microscopy List {Microscopy-at-MSA.Microscopy.Com} 15, 21 -- From: Radice Gary {gradice-at-richmond.edu} 15, 21 -- Subject: S waves in phase contrast optics 15, 21 -- Date: Sun, 11 Feb 2007 15:35:24 -0500 15, 21 -- X-Mailer: Apple Mail (2.752.2) 15, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED 15, 21 -- X-Scanned-By: MIMEDefang 2.49 on 141.166.24.28 ==============================End of - Headers==============================
Another alternative that is to encapsulate in alginate, we have used this to encapsulate both individual cells and tissues. The advantage is that you work at ambient temperature (no heating), disadvantage that you introduce calcium ions into the system which you may not want to do). One method we used was to use a 2% solution of sodium alginate and solidify by dropping into or flooding with 50 mM Calcium Chloride.
Ian
Ian Hallett Microscopy HortResearch Mt Albert Research Centre Private Bag 92 169, Mt Albert Auckland, New Zealand +64-9-815 4200 ext 7002
-----Original Message----- X-from: Gerd.Leitinger-at-meduni-graz.at [mailto:Gerd.Leitinger-at-meduni-graz.at] Sent: Saturday, 10 February 2007 9:35 p.m. To: Ian Hallett
Hi,
When producing Vibratome sections (for EM or light microscopy) of insect tissue, I usually embed the tissue in 15% gelatine before sectioning, and have been doing that for years. But now I suddenly have grown tired of the sticky gelatinous mass that surrounds these fragile sections. Agarose is supposed to be a good substitute- has anyone got experience with it or can tell me which type/temperature/concentration of agarose to test?
thank you
Gerd Leitinger
Dr Gerd Leitinger Institute of Cell Biology, Histology, and Embryology Center of Molecular Medicine Medical University of Graz Harrachgasse 21 8010 Graz Austria
phone +43 316 380 4237
==============================Original Headers============================== 6, 18 -- From Gerd.Leitinger-at-meduni-graz.at Sat Feb 10 02:31:33 2007 6, 18 -- Received: from viefep15-int.chello.at (viefep18-int.chello.at [213.46.255.21]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1A8VWd7009097 6, 18 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 02:31:32 -0600 6, 18 -- Received: from [84.115.150.93] by viefep15-int.chello.at 6, 18 -- (InterMail vM.6.01.05.04 201-2131-123-105-20051025) with ESMTP 6, 18 -- id {20070210083130.VLZ21314.viefep15-int.chello.at-at-[84.115.150.93]} 6, 18 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 09:31:30 +0100 6, 18 -- Message-ID: {45CD82E0.8020407-at-meduni-graz.at} 6, 18 -- Date: Sat, 10 Feb 2007 09:31:28 +0100 6, 18 -- From: "Dr. Gerd Leitinger" {Gerd.Leitinger-at-meduni-graz.at} 6, 18 -- Reply-To: Gerd.Leitinger-at-meduni-graz.at 6, 18 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 6, 18 -- MIME-Version: 1.0 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- Subject: Vibratome sections 6, 18 -- Content-Type: text/plain; charset=ISO-8859-15; format=flowed 6, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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==============================Original Headers============================== 20, 30 -- From IHallett-at-hortresearch.co.nz Sun Feb 11 14:35:51 2007 20, 30 -- Received: from hortresearch.co.nz (mscan.hortresearch.co.nz [202.36.134.15]) 20, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1BKZolx015182 20, 30 -- for {microscopy-at-microscopy.com} ; Sun, 11 Feb 2007 14:35:50 -0600 20, 30 -- Received: from aklexf01.hort.net.nz ([10.16.1.14]) by hortresearch.co.nz 20, 30 -- with HortResearch; Mon, 12 Feb 2007 09:50:28 +1300 20, 30 -- Received: from AKLEXB01.hort.net.nz ([10.16.1.15]) by aklexf01.hort.net.nz 20, 30 -- with Microsoft SMTPSVC(6.0.3790.1830); Mon, 12 Feb 2007 09:35:47 +1300 20, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 20, 30 -- Content-class: urn:content-classes:message 20, 30 -- MIME-Version: 1.0 20, 30 -- Content-Type: text/plain; 20, 30 -- charset=us-ascii 20, 30 -- Subject: RE: [Microscopy] Vibratome sections 20, 30 -- Date: Mon, 12 Feb 2007 09:35:47 +1300 20, 30 -- Message-ID: {D3BAD63C088F3C48AEB385E881359F8702BEFCDD-at-AKLEXB01.hort.net.nz} 20, 30 -- In-Reply-To: {200702100835.l1A8ZSmq013232-at-ns.microscopy.com} 20, 30 -- X-MS-Has-Attach: 20, 30 -- X-MS-TNEF-Correlator: 20, 30 -- Thread-Topic: [Microscopy] Vibratome sections 20, 30 -- Thread-Index: AcdM7myYI71rMw3nTs2gTlPjonwOeABLP43Q 20, 30 -- From: "Ian Hallett" {IHallett-at-hortresearch.co.nz} 20, 30 -- To: {microscopy-at-microscopy.com} 20, 30 -- X-OriginalArrivalTime: 11 Feb 2007 20:35:47.0555 (UTC) 20, 30 -- FILETIME=[3602C330:01C74E1C] 20, 30 -- X-imss-version: 2.046 20, 30 -- X-imss-result: Passed 20, 30 -- X-imss-approveListMatch: *-at-hortresearch.co.nz 20, 30 -- Content-Transfer-Encoding: 8bit 20, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1BKZolx015182 ==============================End of - Headers==============================
It is not that the S wave does not have its phase changed (it does have it's phase changed), but that the D wave is diffracted and the S wave is not diffracted. The diffracted light is also changed in phase by the fact that it is diffracted. So, both change phase (with respect to light that does not pass through the object) due to passing through the object and the D wave has it's phase changed also by being diffracted. The phase plate then puts the D wave and the S wave about 180° out of phase (if it is a positive phase plate) and decreases the intensity of the S wave.
Make sense?
David
On Feb 11, 2007, at 1:40 PM, gradice-at-richmond.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 } ---------------------------------------------------------------------- } ------ } } Here is one of those topics I thought I understood but now that I am } teaching it I find myself confused. How can an S wave pass through a } phase specimen and NOT interact with it? } } I'm using Douglas Murphy's excellent text on light microscopy and } digital imaging. In introducing PC optics he states (pg 99): } } "Upon transit through a phase object, and incident wave of an } illuminating beam becomes divided into two components: (1) an } undeviated (0th order) waved or surround wave (S wave) that passes } through the specimen but does not interact with it, and (2) a } deviated or diffracted wave (D wave) that becomes scattered in many } directions." . } } At the (also excellent) Molecular Expression web site, Murphy and } colleagues say it slightly differently: } } "The primary component is an undeviated (or undiffracted; zeroth- } order) planar wavefront, commonly referred to as the surround (S) } wave, which passes through and around the specimen, but does not } interact with it." } } [http://www.microscopyu.com/articles/phasecontrast/ } phasemicroscopy.html] } } How is it that a light wave can pass through a phase object yet not } interact with the object and not have its phase changed, while the } diffracted light does have its phase changed? } } } Gary Radice } } } } } } } } ==============================Original } Headers============================== } 15, 21 -- From gradice-at-richmond.edu Sun Feb 11 14:35:27 2007 } 15, 21 -- Received: from nylon.richmond.edu (nylon.richmond.edu } [141.166.30.20]) } 15, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l1BKZQxO014910 } 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 } 14:35:26 -0600 } 15, 21 -- Received: from polyester.richmond.edu } (polyester.richmond.edu [141.166.24.28]) } 15, 21 -- by nylon.richmond.edu (8.13.1/8.13.1) with ESMTP id } l1BKZQ2Q009375 } 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 } 15:35:26 -0500 } 15, 21 -- Received: from [10.0.1.3] (lvs080049.richmond.edu } [141.166.80.49]) } 15, 21 -- by polyester.richmond.edu (8.12.11.20060308/8.12.11) } with ESMTP id l1BKZPVs032335 } 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 } 15:35:26 -0500 } 15, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 15, 21 -- Content-Transfer-Encoding: 7bit } 15, 21 -- Message-Id: {79E3462E-2520-4BB2- } BAE0-209724B7F43C-at-richmond.edu} } 15, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } 15, 21 -- To: Microscopy List {Microscopy-at-MSA.Microscopy.Com} } 15, 21 -- From: Radice Gary {gradice-at-richmond.edu} } 15, 21 -- Subject: S waves in phase contrast optics } 15, 21 -- Date: Sun, 11 Feb 2007 15:35:24 -0500 } 15, 21 -- X-Mailer: Apple Mail (2.752.2) } 15, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED } 15, 21 -- X-Scanned-By: MIMEDefang 2.49 on 141.166.24.28 } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 23 -- From Elliott-at-arizona.edu Sun Feb 11 19:15:50 2007 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 l1C1Fn9L009050 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 19:15:50 -0600 9, 23 -- Received: from localhost (amavis6.email.arizona.edu [10.0.0.209]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 862DB12CA8CD 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 18:15:49 -0700 (MST) 9, 23 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9EECE12CAA35 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 18:15:47 -0700 (MST) 9, 23 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 23 -- In-Reply-To: {200702112040.l1BKewGN025217-at-ns.microscopy.com} 9, 23 -- References: {200702112040.l1BKewGN025217-at-ns.microscopy.com} 9, 23 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 23 -- Message-Id: {7D863B6C-E07D-425B-BF5C-30C6B4A65330-at-arizona.edu} 9, 23 -- From: David Elliott {Elliott-at-arizona.edu} 9, 23 -- Subject: Re: [Microscopy] S waves in phase contrast optics 9, 23 -- Date: Sun, 11 Feb 2007 18:15:46 -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 l1C1Fn9L009050 ==============================End of - Headers==============================
A. Diffraction is a simplified means of understanding the aspect of Phase Contrast:
As stated by David, there are phase changes in both waves (S & D); for most biologic specimens on the order of 1/4 wavelength. But (positive) phase plate adds an addition 1/4 wave shift thus creating 180 degree difference where cancellation creates no light (black).
Let me take Chrysotile asbestos fibers at the limit of detection:
B. It is actually better to see phase contrast looking at it in terms of Fourier Optics.
Presume coherent illumination Presume step function phase object
g(y) = e^(i*Phase*y)
Presume a small phase difference
g(y) = 1 + i*Phase*(y)
The Fourier transform is
U(v) = Integral from -infinity to +infinity [1+ (i*Phase*y)]*e^(ivy)
U(v) = U1(v) + iU2(v)
The use of a 1/4 wave phase plate shifts U1 + iU2 to U1 + U2 with a new image function:
g’(y’) = g1(y’) + g2(y’)
Where g1 is the image of the whole object aperture; it represents the constant background;
And g2 is a function for a regular grating.
Phase modulation in the object is converted to an amplitude modulation in the image.
Tony
Ps I have a couple of slides with this if you want to see the equations along with graphics. {From A presentations in 2003}
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-----Original Message----- X-from: Elliott-at-arizona.edu [mailto:Elliott-at-arizona.edu] Sent: Sunday, February 11, 2007 8:22 PM To: ph2-at-sprynet.com
Hi Gary
It is not that the S wave does not have its phase changed (it does have it's phase changed), but that the D wave is diffracted and the S wave is not diffracted. The diffracted light is also changed in phase by the fact that it is diffracted. So, both change phase (with respect to light that does not pass through the object) due to passing through the object and the D wave has it's phase changed also by being diffracted. The phase plate then puts the D wave and the S wave about 180° out of phase (if it is a positive phase plate) and decreases the intensity of the S wave.
Make sense?
David
On Feb 11, 2007, at 1:40 PM, gradice-at-richmond.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 } ---------------------------------------------------------------------- } ------ } } Here is one of those topics I thought I understood but now that I am } teaching it I find myself confused. How can an S wave pass through a } phase specimen and NOT interact with it? } } I'm using Douglas Murphy's excellent text on light microscopy and } digital imaging. In introducing PC optics he states (pg 99): } } "Upon transit through a phase object, and incident wave of an } illuminating beam becomes divided into two components: (1) an } undeviated (0th order) waved or surround wave (S wave) that passes } through the specimen but does not interact with it, and (2) a } deviated or diffracted wave (D wave) that becomes scattered in many } directions." . } } At the (also excellent) Molecular Expression web site, Murphy and } colleagues say it slightly differently: } } "The primary component is an undeviated (or undiffracted; zeroth- } order) planar wavefront, commonly referred to as the surround (S) } wave, which passes through and around the specimen, but does not } interact with it." } } [http://www.microscopyu.com/articles/phasecontrast/ } phasemicroscopy.html] } } How is it that a light wave can pass through a phase object yet not } interact with the object and not have its phase changed, while the } diffracted light does have its phase changed? } } } Gary Radice } } } } } } } } ==============================Original } Headers============================== } 15, 21 -- From gradice-at-richmond.edu Sun Feb 11 14:35:27 2007 } 15, 21 -- Received: from nylon.richmond.edu (nylon.richmond.edu } [141.166.30.20]) } 15, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l1BKZQxO014910 } 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 } 14:35:26 -0600 } 15, 21 -- Received: from polyester.richmond.edu } (polyester.richmond.edu [141.166.24.28]) } 15, 21 -- by nylon.richmond.edu (8.13.1/8.13.1) with ESMTP id } l1BKZQ2Q009375 } 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 } 15:35:26 -0500 } 15, 21 -- Received: from [10.0.1.3] (lvs080049.richmond.edu } [141.166.80.49]) } 15, 21 -- by polyester.richmond.edu (8.12.11.20060308/8.12.11) } with ESMTP id l1BKZPVs032335 } 15, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 } 15:35:26 -0500 } 15, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 15, 21 -- Content-Transfer-Encoding: 7bit } 15, 21 -- Message-Id: {79E3462E-2520-4BB2- } BAE0-209724B7F43C-at-richmond.edu} } 15, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } 15, 21 -- To: Microscopy List {Microscopy-at-MSA.Microscopy.Com} } 15, 21 -- From: Radice Gary {gradice-at-richmond.edu} } 15, 21 -- Subject: S waves in phase contrast optics } 15, 21 -- Date: Sun, 11 Feb 2007 15:35:24 -0500 } 15, 21 -- X-Mailer: Apple Mail (2.752.2) } 15, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED } 15, 21 -- X-Scanned-By: MIMEDefang 2.49 on 141.166.24.28 } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 23 -- From Elliott-at-arizona.edu Sun Feb 11 19:15:50 2007 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 l1C1Fn9L009050 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 19:15:50 -0600 9, 23 -- Received: from localhost (amavis6.email.arizona.edu [10.0.0.209]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 862DB12CA8CD 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 18:15:49 -0700 (MST) 9, 23 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9EECE12CAA35 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 11 Feb 2007 18:15:47 -0700 (MST) 9, 23 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 23 -- In-Reply-To: {200702112040.l1BKewGN025217-at-ns.microscopy.com} 9, 23 -- References: {200702112040.l1BKewGN025217-at-ns.microscopy.com} 9, 23 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 23 -- Message-Id: {7D863B6C-E07D-425B-BF5C-30C6B4A65330-at-arizona.edu} 9, 23 -- From: David Elliott {Elliott-at-arizona.edu} 9, 23 -- Subject: Re: [Microscopy] S waves in phase contrast optics 9, 23 -- Date: Sun, 11 Feb 2007 18:15:46 -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 l1C1Fn9L009050 ==============================End of - Headers==============================
==============================Original Headers============================== 40, 28 -- From ph2-at-sprynet.com Sun Feb 11 19:51:29 2007 40, 28 -- Received: from elasmtp-mealy.atl.sa.earthlink.net (elasmtp-mealy.atl.sa.earthlink.net [209.86.89.69]) 40, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1C1pT72020755 40, 28 -- for {microscopy-at-microscopy.com} ; Sun, 11 Feb 2007 19:51:29 -0600 40, 28 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 40, 28 -- s=dk20050327; d=sprynet.com; 40, 28 -- b=Zt1deRx4127NeyiP8PeX5S5/O59ZCDleMyBzUIzoJBXWqU7K5oA1f4mMKWKDM8nI; 40, 28 -- h=Received:From:To:Cc:Subject:Date:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Mailer:In-Reply-To:X-MimeOLE:Thread-Index:Message-ID:X-ELNK-Trace:X-Originating-IP; 40, 28 -- Received: from [69.136.160.207] (helo=user915fa8f284) 40, 28 -- by elasmtp-mealy.atl.sa.earthlink.net with asmtp (Exim 4.34) 40, 28 -- id 1HGQLl-00029i-Ch; Sun, 11 Feb 2007 20:51:26 -0500 40, 28 -- From: "Tony Havics" {ph2-at-sprynet.com} 40, 28 -- To: "Micrscopy Listserve" {microscopy-at-microscopy.com} 40, 28 -- Cc: {Elliott-at-arizona.edu} 40, 28 -- Subject: RE: [Microscopy] Re: S waves in phase contrast optics 40, 28 -- Date: Sun, 11 Feb 2007 20:51:21 -0500 40, 28 -- MIME-Version: 1.0 40, 28 -- Content-Type: text/plain; 40, 28 -- charset="iso-8859-1" 40, 28 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 40, 28 -- In-Reply-To: {200702120121.l1C1Lq1A017988-at-ns.microscopy.com} 40, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 40, 28 -- Thread-Index: AcdORCzRxXlHaMt2TGavK3TBd2fcyAAAVPxA 40, 28 -- Message-ID: {E1HGQLl-00029i-Ch-at-elasmtp-mealy.atl.sa.earthlink.net} 40, 28 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f911d68cf6ca7668f853a6cacab17c013c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 40, 28 -- X-Originating-IP: 69.136.160.207 40, 28 -- Content-Transfer-Encoding: 8bit 40, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1C1pT72020755 ==============================End of - Headers==============================
It does not really matter, IMO. The submissions are not summaries but rather the whole article. This is counter to other conferences. I'm not willing to put a bunch of time into creating a complete article only to have it rejected for some obscure reason... or to have it relegated to a poster session as before.
I think attendance at M&M is great but paper submission is not an efficient use of my time.
Why they selected Cleveland OH is beyond me. Flight connections from West Coast are lousey.
Try:
http://microscopy.org/MMMeetings/MM07/
for info.
gary g.
At 06:56 PM 2/9/2007, you wrote:
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==============================Original Headers============================== 15, 20 -- From gary-at-gaugler.com Sun Feb 11 20:22:41 2007 15, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 15, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l1C2MeDQ032396 15, 20 -- for {microscopy-at-microscopy.com} ; Sun, 11 Feb 2007 20:22:41 -0600 15, 20 -- Message-Id: {200702120222.l1C2MeDQ032396-at-ns.microscopy.com} 15, 20 -- Received: (qmail 9409 invoked from network); 11 Feb 2007 18:16:36 -0800 15, 20 -- Received: by simscan 1.1.0 ppid: 9405, pid: 9406, t: 0.1775s 15, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 15, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 15, 20 -- by qsmtp4 with SMTP; 11 Feb 2007 18:16:36 -0800 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 15, 20 -- Date: Sun, 11 Feb 2007 18:22:41 -0800 15, 20 -- To: walck-at-southbaytech.com 15, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 15, 20 -- Subject: Re: [Microscopy] Submission deadline 15, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 15, 20 -- In-Reply-To: {200702100256.l1A2uEH1028322-at-ns.microscopy.com} 15, 20 -- References: {200702100256.l1A2uEH1028322-at-ns.microscopy.com} 15, 20 -- Mime-Version: 1.0 15, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-A832892 ==============================End of - Headers==============================
I've got an Hitachi 7000 TEM. There are 3 rotary backing pumps with oil mist filters. Standard Hitachi issue. The time has come to change the filters, but........I can't buy the filters on their own. Instead I'm expected to replace the entire filter casing for almost $300 each! I can't vent the pumps outside, so have to find the filters. The filters are cylindrical, 46.5mm deep, external diameter 64mm, internal diameter 59mm and made of a felty substance. Does anyone know of a source? Down under would be nice. Or from NZ. But at this stage I'm getting desperate and will consider any country on Earth - or other planet if available with short delivery time.
Cheers,
Diana
Diana van Driel Dept Ophthalmology Sydney University GPO Box 4337 Sydney, NSW AUSTRALIA 2001
I'm not sure what these look like but if they are KF25, then look for traps from Duniway Stockroom. They have all sorts of vacuum accessories.
Also, what are the brands of the pumps? Most pump makers offer filters directly. Duniway is likely to be more cost effective.
http://www.duniway.com
gary g.
At 06:53 PM 2/11/2007, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Sun Feb 11 21:51:35 2007 11, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l1C3pZF8023799 11, 20 -- for {microscopy-at-microscopy.com} ; Sun, 11 Feb 2007 21:51:35 -0600 11, 20 -- Message-Id: {200702120351.l1C3pZF8023799-at-ns.microscopy.com} 11, 20 -- Received: (qmail 9611 invoked from network); 11 Feb 2007 19:51:34 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 9608, pid: 9609, t: 0.1692s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp2 with SMTP; 11 Feb 2007 19:51:34 -0800 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 11, 20 -- Date: Sun, 11 Feb 2007 19:51:35 -0800 11, 20 -- To: dianavd-at-eye.usyd.edu.au 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] Hitachi rotary pump oil mist filters 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200702120253.l1C2rJ3J014578-at-ns.microscopy.com} 11, 20 -- References: {200702120253.l1C2rJ3J014578-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-7369F59 ==============================End of - Headers==============================
My mistake. Another conference is in Cleveland. I'm presenting there shortly before M&M.
This M&M is at Ft. Lauderdale., FL. Never been there.
Sorry about that....relative to the venue. Article issues remain.
gary g.
At 06:24 PM 2/11/2007, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Mon Feb 12 02:30:28 2007 11, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l1C8USEO006657 11, 20 -- for {microscopy-at-microscopy.com} ; Mon, 12 Feb 2007 02:30:28 -0600 11, 20 -- Message-Id: {200702120830.l1C8USEO006657-at-ns.microscopy.com} 11, 20 -- Received: (qmail 32246 invoked from network); 12 Feb 2007 00:24:23 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 32243, pid: 32244, t: 0.1874s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp4 with SMTP; 12 Feb 2007 00:24:23 -0800 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 11, 20 -- Date: Mon, 12 Feb 2007 00:30:28 -0800 11, 20 -- To: gary-at-gaugler.com 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] Re: Submission deadline 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200702120224.l1C2OEla002925-at-ns.microscopy.com} 11, 20 -- References: {200702120224.l1C2OEla002925-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-6E226354 ==============================End of - Headers==============================
The College of Microscopy will be offering the following electron microscopy short courses next month:
March 19-23 - Scanning Electron Microscopy
March 27-29 - Transmission Electron Microscopy
Both will be held at our Westmont, IL facility.
In addition to lectures, these courses emphasize hands-on training using state of the art equipment. For further details and registration information, please follow the link below.
www.collegeofmicroscopy.com
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
==============================Original Headers============================== 11, 27 -- From eschumacher-at-mccrone.com Mon Feb 12 08:30:23 2007 11, 27 -- Received: from pgp.mccrone.com (McCrone-Associates-1051626.cust-rtr.ameritech.net [68.23.63.122]) 11, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1CEUNUa003985 11, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 Feb 2007 08:30:23 -0600 11, 27 -- Received: from pgp.mccrone.com (localhost [127.0.0.1]) 11, 27 -- by pgp.mccrone.com (Postfix) with ESMTP id F32631A800B 11, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 Feb 2007 08:30:23 -0600 (CST) 11, 27 -- Received: from MCCRONEMSG.tmg.mccrone.com ([192.168.101.20]) 11, 27 -- by pgp.mccrone.com (PGP Universal service); 11, 27 -- Mon, 12 Feb 2007 08:30:24 -0600 11, 27 -- X-PGP-Universal: processed 11, 27 -- Content-class: urn:content-classes:message 11, 27 -- MIME-Version: 1.0 11, 27 -- Content-Type: text/plain; 11, 27 -- charset="US-ASCII" 11, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 11, 27 -- Subject: Short Course Announcement: SEM and TEM 11, 27 -- Date: Mon, 12 Feb 2007 08:30:12 -0600 11, 27 -- Message-ID: {2A27CE0EC2A5C748974EA513DFB285A7B97435-at-MCCRONEMSG.tmg.mccrone.com} 11, 27 -- X-MS-Has-Attach: 11, 27 -- X-MS-TNEF-Correlator: 11, 27 -- Thread-Topic: Short Course Announcement: SEM and TEM 11, 27 -- Thread-Index: AcdOsk5IwJftz2oOQQ6HFs6FZ8o43g== 11, 27 -- From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} 11, 27 -- To: {Microscopy-at-MSA.Microscopy.Com} 11, 27 -- Content-Transfer-Encoding: 8bit 11, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1CEUNUa003985 ==============================End of - Headers==============================
For EM embedding of loose cells (in order to make them easier to handle), we have used Sigma's Type VII or Type IX agarose. I have not developed a preference, since this is not something i really do routinely. You may or may not like agarose after having used gelatin. While agarose is more clear and also less visible in the microscope, and is probably better for the cryo knife, I have found it more tricky to handle - keeping it solid, etc. The concentrations and references are a bit further away to reach at this moment, but I'll look them up if you haven't gotten them yet from other sources.
Best regards, Vlad
________________________________________________ Vlad Speransky, Staff Scientist Supramolecular Structure and Function Section NIBIB, National Institutes of Health 13 South Dr, Rm. 3N17 MSC 5766 Bethesda, MD 20892 301 496-3989 vladislav_speransky-at-nih.gov
On Feb 10, 2007, at 3:33 AM, Gerd.Leitinger-at-meduni-graz.at 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, } } When producing Vibratome sections (for EM or light microscopy) of } insect } tissue, I usually embed the tissue in 15% gelatine before sectioning, } and have been doing that for years. But now I suddenly have grown } tired } of the sticky gelatinous mass that surrounds these fragile sections. } Agarose is supposed to be a good substitute- has anyone got experience } with it or can tell me which type/temperature/concentration of agarose } to test? } } thank you } } Gerd Leitinger } } Dr Gerd Leitinger } Institute of Cell Biology, Histology, and Embryology } Center of Molecular Medicine } Medical University of Graz } Harrachgasse 21 } 8010 Graz } Austria } } phone +43 316 380 4237 } } ==============================Original } Headers============================== } 6, 18 -- From Gerd.Leitinger-at-meduni-graz.at Sat Feb 10 02:31:33 2007 } 6, 18 -- Received: from viefep15-int.chello.at (viefep18- } int.chello.at [213.46.255.21]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l1A8VWd7009097 } 6, 18 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb 2007 } 02:31:32 -0600 } 6, 18 -- Received: from [84.115.150.93] by viefep15-int.chello.at } 6, 18 -- (InterMail vM.6.01.05.04 } 201-2131-123-105-20051025) with ESMTP } 6, 18 -- id {20070210083130.VLZ21314.viefep15- } int.chello.at-at-[84.115.150.93]} } 6, 18 -- for {microscopy-at-microscopy.com} ; Sat, 10 Feb } 2007 09:31:30 +0100 } 6, 18 -- Message-ID: {45CD82E0.8020407-at-meduni-graz.at} } 6, 18 -- Date: Sat, 10 Feb 2007 09:31:28 +0100 } 6, 18 -- From: "Dr. Gerd Leitinger" {Gerd.Leitinger-at-meduni-graz.at} } 6, 18 -- Reply-To: Gerd.Leitinger-at-meduni-graz.at } 6, 18 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) } 6, 18 -- MIME-Version: 1.0 } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- Subject: Vibratome sections } 6, 18 -- Content-Type: text/plain; charset=ISO-8859-15; format=flowed } 6, 18 -- Content-Transfer-Encoding: 7bit } ==============================End of - } Headers==============================
==============================Original Headers============================== 8, 25 -- From vladislav_speransky-at-nih.gov Mon Feb 12 08:36:57 2007 8, 25 -- Received: from nihrelayxway.hub.nih.gov (nihrelayxway.hub.nih.gov [128.231.90.106]) 8, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1CEauTE011927 8, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Feb 2007 08:36:56 -0600 8, 25 -- Received: from helix.nih.gov ([128.231.2.3]) 8, 25 -- by nihrelayxway.hub.nih.gov with ESMTP; 12 Feb 2007 09:36:56 -0500 8, 25 -- X-IronPortListener: NIH_Relay 8, 25 -- X-SBRS: None 8, 25 -- X-IronPort-AV: i="4.13,314,1167627600"; 8, 25 -- d="scan'208"; a="496577040:sNHT39579976" 8, 25 -- Received: from [156.40.102.124] ([156.40.102.124]) 8, 25 -- by helix.nih.gov (8.13.6/8.11.7/2SCANNER) with ESMTP id l1CEauuQ48952608; 8, 25 -- Mon, 12 Feb 2007 09:36:56 -0500 (EST) 8, 25 -- In-Reply-To: {200702100833.l1A8X6r8010766-at-ns.microscopy.com} 8, 25 -- References: {200702100833.l1A8X6r8010766-at-ns.microscopy.com} 8, 25 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 25 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 25 -- Message-Id: {8D8A3748-1897-49C9-BA85-3FAB226CEC48-at-nih.gov} 8, 25 -- Cc: Microscopy-at-microscopy.com 8, 25 -- Content-Transfer-Encoding: 7bit 8, 25 -- From: Vlad Speransky {vladislav_speransky-at-nih.gov} 8, 25 -- Subject: Re: [Microscopy] Vibratome sections 8, 25 -- Date: Mon, 12 Feb 2007 09:35:45 -0500 8, 25 -- To: Gerd.Leitinger-at-meduni-graz.at 8, 25 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
The M&M procedure is actually easier than just about anyone else's. You write a single page of text, a couple of hundred words, add a page of pictures and you are done. As opposed to writing an abstract and then, months later, writing a much longer paper. Those of us who had to go through a clearance procedure at our institution had much less than half the work.
As far as risking rejection, the M&M program production meeting, where each article is read by at least two scientists, has a very low rejection rate. Papers are rejected for REALLY bad science, fraud, gross non-compliance with the simple format rules. etc. In no way does a low rejection rate equate to bad science getting through. The program committee people are first-rate.
There are a finite number of platform presentation rooms available, which translates to a fixed number of possible platform presentation slots. The program committee members organize symposia and invite platform presentation speakers. There are hundreds of submissions above and beyond the number of presentation slots available. It follows that a lot of good articles are presented as posters. Posters are a very effective way of presenting results. In no way are posters "second rate."
Cleveland?? We met there in the last century! We chose Cleveland because there is a strong microscopy community in the area who worked hard to bring M&M to Ohio. Difficult flight connections from the West Coast? Does this mean you had to change planes in Chicago? Changing planes is what most East Coast people have to do to get to great M&M cities like Portland OR.
M&M this year is in Fort Lauderdale, Florida. A big meeting is expected in a beautiful convention center with pre- and post-convention vacation options in one of the world's greatest fun locations. Please join us--the ocean is warm.
Ron
gary-at-gaugler.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 } ---------------------------------------------------------------------------- } } It does not really matter, IMO. The submissions are } not summaries but rather the whole article. This is } counter to other conferences. I'm not willing to } put a bunch of time into creating a complete article } only to have it rejected for some obscure reason... } or to have it relegated to a poster session as before. } } I think attendance at M&M is great but paper submission } is not an efficient use of my time. } } Why they selected Cleveland OH is beyond me. Flight } connections from West Coast are lousey. } } Try: } } http://microscopy.org/MMMeetings/MM07/ } } for info. } } } gary g. } } } At 06:56 PM 2/9/2007, you 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 website for the M&M 2007 meeting says that the submission deadline is } } Wed Feb 15, 2007. Feb 15 is a Thursday on my calendar. Can anybody set me } } straight? I can't find my call for papers. } } } } Thanks in advance } } } } -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 Headers============================== } } 4, 20 -- From walck-at-southbaytech.com Fri Feb 9 20:55:07 2007 } } 4, 20 -- Received: from flpi102.sbcis.sbc.com (flpi102.sbcis.sbc.com } } [207.115.20.71]) } } 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l1A2t78m025426 } } 4, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 } } 20:55:07 -0600 } } 4, 20 -- X-ORBL: [64.169.217.123] } } 4, 20 -- Received: from dynamicbl8uno3 } } (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) } } 4, 20 -- by flpi102.sbcis.sbc.com (8.13.8 } } out.dk.spool/8.13.8) with ESMTP id l1A2t5JY025273 } } 4, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 } } 18:55:06 -0800 } } 4, 20 -- From: "Scott Walck" {walck-at-southbaytech.com} } } 4, 20 -- To: {Microscopy-at-microscopy.com} } } 4, 20 -- Subject: Submission deadline } } 4, 20 -- Date: Fri, 9 Feb 2007 18:55:15 -0800 } } 4, 20 -- Message-ID: {005a01c74cbe$e4ba99c0$7801a8c0-at-dynamicbl8uno3} } } 4, 20 -- MIME-Version: 1.0 } } 4, 20 -- Content-Type: text/plain; } } 4, 20 -- charset="us-ascii" } } 4, 20 -- Content-Transfer-Encoding: 7bit } } 4, 20 -- X-Mailer: Microsoft Office Outlook 11 } } 4, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 } } 4, 20 -- Thread-Index: AcdMvuQfw00QSO25QAWK7gwDn7jIfg== } } ==============================End of - Headers============================== } } } } } ==============================Original Headers============================== } 15, 20 -- From gary-at-gaugler.com Sun Feb 11 20:22:41 2007 } 15, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) } 15, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l1C2MeDQ032396 } 15, 20 -- for {microscopy-at-microscopy.com} ; Sun, 11 Feb 2007 20:22:41 -0600 } 15, 20 -- Message-Id: {200702120222.l1C2MeDQ032396-at-ns.microscopy.com} } 15, 20 -- Received: (qmail 9409 invoked from network); 11 Feb 2007 18:16:36 -0800 } 15, 20 -- Received: by simscan 1.1.0 ppid: 9405, pid: 9406, t: 0.1775s } 15, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 } 15, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } 15, 20 -- by qsmtp4 with SMTP; 11 Feb 2007 18:16:36 -0800 } 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 } 15, 20 -- Date: Sun, 11 Feb 2007 18:22:41 -0800 } 15, 20 -- To: walck-at-southbaytech.com } 15, 20 -- From: Gary Gaugler {gary-at-gaugler.com} } 15, 20 -- Subject: Re: [Microscopy] Submission deadline } 15, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} } 15, 20 -- In-Reply-To: {200702100256.l1A2uEH1028322-at-ns.microscopy.com} } 15, 20 -- References: {200702100256.l1A2uEH1028322-at-ns.microscopy.com} } 15, 20 -- Mime-Version: 1.0 } 15, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-A832892 } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 10, 19 -- From randerson20-at-tampabay.rr.com Mon Feb 12 08:53:31 2007 10, 19 -- Received: from ms-smtp-07.tampabay.rr.com (ms-smtp-07.tampabay.rr.com [65.32.5.139]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1CErUfO026852 10, 19 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Feb 2007 08:53:30 -0600 10, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 10, 19 -- by ms-smtp-07.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id l1CErQAT022825; 10, 19 -- Mon, 12 Feb 2007 09:53:28 -0500 (EST) 10, 19 -- Message-ID: {45D07F64.2030300-at-tampabay.rr.com} 10, 19 -- Date: Mon, 12 Feb 2007 09:53:24 -0500 10, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 10, 19 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) 10, 19 -- MIME-Version: 1.0 10, 19 -- To: gary-at-gaugler.com, Listserver {Microscopy-at-Microscopy.Com} 10, 19 -- Subject: Re: [Microscopy] Re: Submission deadline 10, 19 -- References: {200702120222.l1C2MpDo032599-at-ns.microscopy.com} 10, 19 -- In-Reply-To: {200702120222.l1C2MpDo032599-at-ns.microscopy.com} 10, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 19 -- Content-Transfer-Encoding: 7bit 10, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
A question regarding the on-line submission for M&M 2007: I submitted my paper last week. During the process, I was told for a couple of times that there would be an email sent to me either for checking file or as a recipt of completion of submission. However, I haven't yet received any email. Now I start to worry about this because it was also said an email to confirm my student status would be sent to my advisor(abroad now), but I dont believe he will get the email either. I also emailed the "support-at-bono.cup.org" and had no reply. Anyone experienced can tell me what to do? Since the deadline is approaching, I really really appreciate your prompt reply.
Fan
==============================Original Headers============================== 5, 19 -- From fli-at-chem.umn.edu Mon Feb 12 09:48:25 2007 5, 19 -- Received: from chem.umn.edu (chemsun.chem.umn.edu [128.101.157.2]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l1CFmPaD007090 5, 19 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Feb 2007 09:48:25 -0600 5, 19 -- Message-Id: {200702121548.l1CFmPaD007090-at-ns.microscopy.com} 5, 19 -- Received: (qmail 30107 invoked from network); 12 Feb 2007 15:48:24 -0000 5, 19 -- Received: from x160-1-dhcp.chem.umn.edu (HELO fan) (128.101.160.1) 5, 19 -- by 0 with SMTP; 12 Feb 2007 15:48:24 -0000 5, 19 -- From: "Fan Li" {fli-at-chem.umn.edu} 5, 19 -- To: {Microscopy-at-microscopy.com} 5, 19 -- Subject: Question-M&M 07 paper submission 5, 19 -- Date: Mon, 12 Feb 2007 09:48:17 -0600 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: text/plain; 5, 19 -- charset="us-ascii" 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 5, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 5, 19 -- Thread-Index: AcdOvTaVcIpox5O3Tu2XgM8Rr9l07w== ==============================End of - Headers==============================
I came across a short article in a British PC magazine (PC Format January 2007) that described combining an old Watcom graphics tablet with a 15 inch LCD screen to produce a cheap pen display system like the Watcom Cintiq. No idea how well it works in practice.
Ian
Ian Hallett Microscopy HortResearch Mt Albert Research Centre Private Bag 92 169, Mt Albert Auckland, New Zealand +64-9-815 4200 ext 7002
-----Original Message----- X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu] Sent: Saturday, 10 February 2007 3:48 p.m. To: Ian Hallett
Microscopy folks,
I'm looking for the most efficient and accurate way to trace features in digital images (for instance, outlining different minerals or metal phases in backscattered electron images). Does anyone have experience with this? There seems to be three basic ways to go: (1) graphics pads that replace a mouse, (2) Tablet PCs, and (3) "pen displays" -- that is, LCD displays on which, like Tablet PCs, a stylus can write on the screen. Any relevant experiences out there with such devices? Any insights would be welcome. Feel free to email me off-listserver with any comments or suggestions. If there is a demand, I can post a summary of the answers.
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu Personal Website: http://umn.edu/~frah0010
==============================Original Headers============================== 8, 20 -- From frah0010-at-umn.edu Fri Feb 9 20:44:31 2007 8, 20 -- Received: from mta-m2.tc.umn.edu (mta-m2.tc.umn.edu [160.94.23.21]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1A2iUKV014238 8, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Feb 2007 20:44:31 -0600 8, 20 -- Received: from [10.0.1.3] (c-75-72-182-252.hsd1.mn.comcast.net [75.72.182.252]) 8, 20 -- by mta-m2.tc.umn.edu (UMN smtpd) with ESMTP 8, 20 -- Fri, 9 Feb 2007 20:44:29 -0600 (CST) 8, 20 -- X-Umn-Remote-Mta: [N] c-75-72-182-252.hsd1.mn.comcast.net [75.72.182.252] #+TS+AU+HN 8, 20 -- In-Reply-To: {1D190879-3592-4F6F-B0BE-857EB4840C8A-at-umn.edu} 8, 20 -- References: {1D190879-3592-4F6F-B0BE-857EB4840C8A-at-umn.edu} 8, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 20 -- Message-Id: {A3D8A22D-6E52-41EA-925F-6FACADA2DB4C-at-umn.edu} 8, 20 -- Cc: Ellery Frahm {frah0010-at-umn.edu} 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- From: Ellery Frahm {frah0010-at-umn.edu} 8, 20 -- Subject: digital sketching 8, 20 -- Date: Fri, 9 Feb 2007 20:44:22 -0600 8, 20 -- To: Microscopy-at-microscopy.com 8, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
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==============================Original Headers============================== 21, 30 -- From IHallett-at-hortresearch.co.nz Mon Feb 12 14:04:51 2007 21, 30 -- Received: from hortresearch.co.nz (mscan.hortresearch.co.nz [202.36.134.15]) 21, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1CK4ore025189 21, 30 -- for {microscopy-at-microscopy.com} ; Mon, 12 Feb 2007 14:04:50 -0600 21, 30 -- Received: from aklexf01.hort.net.nz ([10.16.1.14]) by hortresearch.co.nz 21, 30 -- with HortResearch; Tue, 13 Feb 2007 09:19:30 +1300 21, 30 -- Received: from AKLEXB01.hort.net.nz ([10.16.1.15]) by aklexf01.hort.net.nz 21, 30 -- with Microsoft SMTPSVC(6.0.3790.1830); Tue, 13 Feb 2007 09:04:49 +1300 21, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 21, 30 -- Content-class: urn:content-classes:message 21, 30 -- MIME-Version: 1.0 21, 30 -- Content-Type: text/plain; 21, 30 -- charset=us-ascii 21, 30 -- Subject: RE: [Microscopy] digital sketching 21, 30 -- Date: Tue, 13 Feb 2007 09:04:48 +1300 21, 30 -- Message-ID: {D3BAD63C088F3C48AEB385E881359F8702BEFFAD-at-AKLEXB01.hort.net.nz} 21, 30 -- In-Reply-To: {200702100247.l1A2lgu2018321-at-ns.microscopy.com} 21, 30 -- X-MS-Has-Attach: 21, 30 -- X-MS-TNEF-Correlator: 21, 30 -- Thread-Topic: [Microscopy] digital sketching 21, 30 -- Thread-Index: AcdMvde7NGXenUr4RgCyKhyqnhZCDwCIbHIg 21, 30 -- From: "Ian Hallett" {IHallett-at-hortresearch.co.nz} 21, 30 -- To: {microscopy-at-microscopy.com} 21, 30 -- X-OriginalArrivalTime: 12 Feb 2007 20:04:49.0083 (UTC) 21, 30 -- FILETIME=[0CB030B0:01C74EE1] 21, 30 -- X-imss-version: 2.046 21, 30 -- X-imss-result: Passed 21, 30 -- X-imss-approveListMatch: *-at-hortresearch.co.nz 21, 30 -- Content-Transfer-Encoding: 8bit 21, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1CK4ore025189 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both sswaffe-at-abv.bg as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: V. Andreev
Organization: NPMG
Title-Subject: [Filtered] Electron microscopy specimen processing
Question: I'm a student, but once I took a trip to the Bulgarian Academy of Sciences and in the Institut of parasitology and experimental pathology I saw the procedure of specimen processing for electron microscopy. The specimen was placed in a capsule (like some antibiotics are) made from a solid resin. Then this capsule was placed into an ultramicrotome and was cut under a magnifying glass. The staff there told me that they were determinating the thickness of the sliced specimen by the color that it gives of (it was somewere about 50-60nm). The questions I propose are two: what is this resin and how are they determinating the thickness by the color?
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Email: DLowry-at-asu.edu Name: David Lowry
Organization: Arizona State University
Title-Subject: [Filtered] sputter-coater
Question: we have an older Technics Hummer sputter-coater. Lately, when operated at the recommended settings arcing occurs during sputtering. I have cleaned the unit as well as possible (I think) but the problem persists. The arcing is mitigated by reducing amperage, but as a result samples require extended time to adequately coat. Currently a Pt target is installed which is in good condition. I was wondering if this is just a consequence of age and I need to settle for using lower mA and longer time to get good sputtering, or if there is another potential cause I have not considered. Thanks
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Question: The Florida Society for Microscopy and the Florida Chapter of the AVS Science and Technology Society will be offering a series of Hands-on Short Courses at their 2007 Joint Annual Symposium:
Rutherford Backscatter Spectroscopy and Secondary Ion Mass Spectroscopy on March 13 Scanning Electron Microscopy and Energy Dispersive Spectroscopy on March 14 & 15 Auger Electron Spectroscopy and X-Ray Photoelectron Spectroscopy March 16
More information and registration at www.flavs.org or contact Amelia Dempere at ldemp-at-mse.ufl.edu
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Email: lh03-at-aub.edu.lb Name: Lucia Hanna
Organization: American University of beirut
Title-Subject: [Filtered] Training courses in Microscopy
Question: Hello,
I work as a research assistant in Plant pathology at the American University of Beirut, Beirut, Lebanon.
Our lab is equipped with a Zeiss Axiophot compound microscope on which I acquired hands on experience in light microscopy, phase contrast, darkfield and fluoresence microscopy. I would like to deepen my knowledge in the various other fileds of microscopy including electron microscopy, live cell imaging and image analysis as applied to the field of Plant Pathology. I would appreciate your help in helping me find appropriate training courses and funding for these courses, specially in toronto, Canada or in the USA , in states close to Toronto Canada.
THE TEXAS SOCIETY FOR MICROSCOPY Spring Meeting April 12-14, 2007 TCU Campus Fort Worth, TX USA
Thursday, April 12, Workshop: “Basic confocal, wave optics and point spread functions” Charles Hemphill, Sponsored by Leica Microsystems Held at TCU campus Kelly Alumni Center
Guest Speakers – Friday, April 13, 2007
Dr. Edward Kolesar Department of Engineering (Nanotechology) TCU, Fort Worth, TX
— An Eyeball and an Engineer — What could they possibly have in common?
This talk will address the implications of understanding human visual accommodation from the perspective of using microelectromechanical systems (MEMS) technology to supplement cilliary muscle involvement.
and
Dr. John P. Janovec BRIT (botanical research institute of Texas) Fort Worth, TX will speak on his research in Peru
See our web site, http://www.texasmicroscopy.org/ for registration forms and hotel details.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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============================== 13, 22 -- From r-holdford-at-ti.com Mon Feb 12 15:41:04 2007 13, 22 -- Received: from arroyo.ext.ti.com (arroyo.ext.ti.com [192.94.94.40]) 13, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1CLf4UK018231 13, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 Feb 2007 15:41:04 -0600 13, 22 -- Received: from dlep33.itg.ti.com ([157.170.170.112]) 13, 22 -- by arroyo.ext.ti.com (8.13.7/8.13.7) with ESMTP id l1CLexl5001877 13, 22 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 13, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 Feb 2007 15:41:04 -0600 13, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 13, 22 -- by dlep33.itg.ti.com (8.13.7/8.13.7) with ESMTP id l1CLewZU017670 13, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 Feb 2007 15:40:58 -0600 (CST) 13, 22 -- Message-ID: {45D0DEEA.3070603-at-ti.com} 13, 22 -- Date: Mon, 12 Feb 2007 15:40:58 -0600 13, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 13, 22 -- Organization: SC Packaging Development -- FA Development 13, 22 -- User-Agent: Thunderbird 1.5.0.9 (Windows/20061207) 13, 22 -- MIME-Version: 1.0 13, 22 -- To: MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 13, 22 -- Subject: First Call for Papers for the Texas Society for Microscopy Spring 13, 22 -- 2007 Meeting 13, 22 -- Content-Type: text/plain; charset=windows-1252; format=flowed 13, 22 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
The colors are a result of interference in the light waves that are reflected from the thin slice of resin. When white light is shined on the slice, some of the light is reflected from the surface, while other light passes through to the opposite surface of the section, then is reflected back up and joins the previously mentioned reflected light, but since it has traveled a little farther (through the thickness of the slice) its waveform is out of phase with the other light. You have probably studied wave interference in physics, so you know that you can have positive (=brightening) interference (when two peaks coincide) or negative (=darkening) interference (when a trough coincides with a peak). Electron microscopists have learned to accurately determine the thickness of their slices by observing the color of the light reflected off the surface (silver and gold are two 'thicknesses' commonly talked about). There are many different kinds of resins which are used to support the specimens (from the inside and the outside). They go by brand names, but chemically they are things like methacrylates, polyesters, acrylics, and epoxies. Before the specimens are infiltrated with and embedded in the resin, there are other important steps that must be done to biological specimens. Different kinds of chemicals are used to 'fix' the specimen, based on which kinds of structures or macromolecules are to be preserved without distortion. A dehydration step is needed to replace water with a different solvent which is compatible with the resin of choice. There are different kinds of electron-dense stains which can be used to increase contrast in parts of the specimen to be examined or to bind to specific structures. There are other things that must sometimes be done as well, so specimen preparation for transmission electron microscopy (TEM) is time-consuming and sometimes quite complicated. Which is why I stick to scanning electron microscopy (SEM) ;o)
Paul
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Email: sswaffe-at-abv.bg Name: V. Andreev
Organization: NPMG
Title-Subject: [Filtered] Electron microscopy specimen processing
Question: I'm a student, but once I took a trip to the Bulgarian Academy of Sciences and in the Institut of parasitology and experimental pathology I saw the procedure of specimen processing for electron microscopy. The specimen was placed in a capsule (like some antibiotics are) made from a solid resin. Then this capsule was placed into an ultramicrotome and was cut under a magnifying glass. The staff there told me that they were determinating the thickness of the sliced specimen by the color that it gives of (it was somewere about 50-60nm). The questions I propose are two: what is this resin and how are they determinating the thickness by the color?
-------------------------------------------------------------------- Few people at the beginning of the nineteenth century needed an adman to tell them what they wanted. - John Kenneth Galbraith
==============================Original Headers============================== 11, 20 -- From pbgrover-at-yahoo.com Mon Feb 12 16:56:24 2007 11, 20 -- Received: from web34212.mail.mud.yahoo.com (web34212.mail.mud.yahoo.com [66.163.178.127]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l1CMuN7r030610 11, 20 -- for {microscopy-at-microscopy.com} ; Mon, 12 Feb 2007 16:56:23 -0600 11, 20 -- Received: (qmail 2192 invoked by uid 60001); 12 Feb 2007 22:56:23 -0000 11, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 20 -- s=s1024; d=yahoo.com; 11, 20 -- h=X-YMail-OSG:Received:Date:From:Reply-To:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 11, 20 -- b=27lWRmvMuT8NaZAks+bZjdjSZi13Q01SJynL8NJFd/uxMNka4YIg6pfI6FFt3sBc3IAdyv7xW4v8Kpff0Xycnqs/X/gD8mCkB7uZPbDvz3BDXjzNLphrdRWxpuhX4ckyp0tAvgD5ISKt1hsUO2gbSZY1LbdwKtMCHU32BviSvXw=; 11, 20 -- X-YMail-OSG: T88paHMVM1n.sFqslxO8843KQZOUBk5.25eMTQIjYX6jd2IF7hMzFOQeWZHo7T4kWmV9ya_ui5ibMhsaTTHv7fqFjzNvL_17l99KJknNPzeQz1mJLK6nx96I403X8VUVhZlpntY6K.6v4G8- 11, 20 -- Received: from [74.140.104.200] by web34212.mail.mud.yahoo.com via HTTP; Mon, 12 Feb 2007 14:56:22 PST 11, 20 -- Date: Mon, 12 Feb 2007 14:56:22 -0800 (PST) 11, 20 -- From: paul grover {pbgrover-at-yahoo.com} 11, 20 -- Reply-To: pbgrover-at-yahoo.com 11, 20 -- Subject: electron microscopy specimen processing 11, 20 -- To: microscopy-at-microscopy.com 11, 20 -- MIME-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=iso-8859-1 11, 20 -- Content-Transfer-Encoding: 8bit 11, 20 -- Message-ID: {987643.193.qm-at-web34212.mail.mud.yahoo.com} ==============================End of - Headers==============================
(Sorry that this is late but I had some trouble with attachments appearing from nowhere and the message being bumped from the list. I persist only because anyone seriously interested in CPD should read Hans Ris' paper.)
Anderson used amyl acetate after ethanol in the first paper in the 1950s, mostly so he could be sure that all of it had been replaced by CO2 before he went through the critical point. Using his original equipment, I can attest that this could take a long time. The AA used to get forced up into the stem of the pressure gauge and took forever to be washed out.
Ethanol and Acetone work fine (if you ignore the unavoidable shrinkage mentioned in a previous post on dimensional measurements). The main difference is how their slightly different densities (with respect to liquid CO2), affects mixing. The bottom line is to use agitation (either by using a magnetic stirrer or by shaking the entire drier) as described by Hans Ris many years ago.
Ris, H, (1985) The cytoplasmic filament system in critical point dried whole mounts and plastic-embedded sections, J.Cell Biol. 100:1474-1487
His other main point was the absolute necessity of making sure that no water remained in the final CO2 transition liquid. As both ethanol and acetone love to pick up water from the air and, as biological tissue loves water even more, you must use molecular sieve to dry both the final ethanol and the liquid CO2 (with a high-pressure filter sold by several manufacturers). Then put the resulting specimen in a desiccator over phosphorous pentoxide.
If you think that parts/million CO2 is dry enough, I suggest that you work our what "concentration" your cells are in the bomb (volume of dry cellular material/volume of bomb). Admittedly, this is more a factor when drying a monolayer on a few EM grids, as he was.
Ris was able to show that, on a number of purified fibrous structures (MTs, chromosomes, Intermediate and actin filaments) failing to use such a filter destroyed their (known) shape. By viewing whole-mount cells in the HVEM he could show that the cytoskeleton was converted into formless "trabeculae".
This paper should be read by anyone planning to use CPD for high-resolution SEM or TEM studies.
Cheers,
Jim Pawley
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I've also used ethanol as the transfer fluid with liquid CO2 for many years. Both Polaron and Ladd CPD's have been used without any problems. I tried amyl acetate for awhile but wasn't too fond of it. It's only advantage was being able to smell minute residues of it during the CPD flushing process.
However, the number of flushes of liquid CO2 and length of time between flushes depends on the size or thickness of your sample. Most of the ethanol is removed in the first few flushes. You really have to become familar with your samples to determine the time required to remove all of the ethanol. And as usual add a few flushes for good measure.
I've found certain arthropods/insects etc. to be most difficult using CPD. The hard exoskeleton or cuticle allows ethanol to enter the body of the organism but doesn't easily allow the exchange of ethanol with liquid CO2. Tardigrades were especially aggravating. Alternative drying techniques than CPD with arthropods and insects are welcome. -- ********************************************** 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 17-28, 2007, UBC, Vancouver Canada Info: http:// www.3dcourse.ubc.ca/ Applications due by March 15, 2007 "If it ain't diffraction, it must be statistics." Anon.
==============================Original Headers============================== 15, 25 -- From jbpawley-at-wisc.edu Tue Feb 13 08:10:38 2007 15, 25 -- Received: from smtpauth.wiscmail.wisc.edu (agogare.doit.wisc.edu [144.92.197.211]) 15, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1DEAcdb028469 15, 25 -- for {Microscopy-at-Microscopy.Com} ; Tue, 13 Feb 2007 08:10:38 -0600 15, 25 -- Received: from avs-daemon.smtpauth2.wiscmail.wisc.edu by 15, 25 -- smtpauth2.wiscmail.wisc.edu 15, 25 -- (Sun Java System Messaging Server 6.2-7.05 (built Sep 5 2006)) 15, 25 -- id {0JDE00L01NDPFE00-at-smtpauth2.wiscmail.wisc.edu} for 15, 25 -- Microscopy-at-Microscopy.Com; Tue, 13 Feb 2007 08:10:37 -0600 (CST) 15, 25 -- Received: from [144.92.238.207] by smtpauth2.wiscmail.wisc.edu 15, 25 -- (Sun Java System Messaging Server 6.2-7.05 (built Sep 5 2006)) 15, 25 -- with ESMTPSA id {0JDE00LAMNDM1600-at-smtpauth2.wiscmail.wisc.edu} for 15, 25 -- Microscopy-at-Microscopy.Com; Tue, 13 Feb 2007 08:10:34 -0600 (CST) 15, 25 -- Date: Tue, 13 Feb 2007 08:10:35 -0600 15, 25 -- From: James Pawley {jbpawley-at-wisc.edu} 15, 25 -- Subject: [Microscopy] RE: viaWWW: critical point drying question 15, 25 -- To: Microscopy-at-Microscopy.Com 15, 25 -- Message-id: {p06240806c1f7759f270b-at-[144.92.238.207]} 15, 25 -- MIME-version: 1.0 15, 25 -- Content-type: text/plain; charset=us-ascii; format=flowed 15, 25 -- Content-transfer-encoding: 7BIT 15, 25 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=144.92.238.207 15, 25 -- X-Spam-PmxInfo: Server=avs-11, Version=5.3.0.289146, 15, 25 -- Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.2.13.55433, 15, 25 -- SenderIP=144.92.238.207 ==============================End of - Headers==============================
I would second the previous comments about using a graphics tablet. We have an inexpensive Wacom Intuos tablet that we have been using for years for manually editing images. We analyze air voids in concrete and computerized processing according to a script goes a long way toward isolating the air voids, but we often encounter voids that were not completely thresholded or false air voids, (e.g., pores in aggregate instead of paste). At that stage, some manual editing is warranted.
The tablet we have offers two modes of movement of the stylus. One mode operates as a mouse with relative motion. The only advantage of that mode over a regular mouse might be more precise control of the stylus as compared to a mouse. The other mode uses absolute positioning. You don't have to worry about the speed of movement; simply moving the stylus from point A to point B will move the cursor from point A to B. Our pad has only a 5-inch active area, but I find it fairly easy to move the stylus on the pad and edit the image on the screen because of the absolute correspondence.
That said, a lot of the usability will depend on the software package. We also employ backscattered imaging for much of our work since phases often stand out well from one another in polished sections and we can employ global thresholding to pick them out. I wonder how your application is different.
I have always used a simple agar embedment for sectioning various tissues with a Vibratome. I start by heating and stirring 4% agar by weight in water until it all goes into solution. Usually some water will evaporate thus the percentage is actually higher than 4% but that is not critical as long as hardens sufficiently when cooled. I store it in a 50ml tube at 4C than melt it by placing it in a beaker of water on a hot plate. To embed the ttissue I put a drop of the agar on a glass microscope slide and then add the tissue and another drop or two of agar as needed to cover the tissue. Then I quickly cool the slide/agar/tissue by placing it on ice. When the agar is hardened after a few minutes I trim the excess agar away with a razor blade and glue the block to the Vibratome stage with cyanoacrylate (SuperGlue).
There are many variations including the use of expensive low temperature melting point agarose or agar for temperature sensitive samples. There are a very confusing number of agar and agarose products offered for sale and I was shocked that some don't behave "correctly" for this purpose. One product would never gel. I get the most ordinary and inexpensive product. My most recent purchase was from Fisher Scientific for Agar # BP1423-500. -- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 4, 28 -- From Larry.Ackerman-at-ucsf.edu Tue Feb 13 14:57:11 2007 4, 28 -- Received: from emfmcb01.ucsfmedicalcenter.org (emfmcb01.ucsfmedicalcenter.org [64.54.46.97]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1DKvAB3028615 4, 28 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Feb 2007 14:57:10 -0600 4, 28 -- Received: from 64.54.128.152 by emfmcb01.ucsfmedicalcenter.org with 4, 28 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 4, 28 -- Tue, 13 Feb 2007 13:05:45 -0800 4, 28 -- X-Server-Uuid: E2E48A14-EE5B-4280-A138-188440602EDD 4, 28 -- Received: from [128.218.123.88] ([128.218.123.88]) by 4, 28 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Tue, 13 Feb 4, 28 -- 2007 12:55:26 -0800 4, 28 -- Message-ID: {45D225BD.8000703-at-ucsf.edu} 4, 28 -- Date: Tue, 13 Feb 2007 12:55:25 -0800 4, 28 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 4, 28 -- Reply-to: larry.ackerman-at-ucsf.edu 4, 28 -- Organization: UCSF, NeuroAnatomy 4, 28 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 4, 28 -- X-Accept-Language: en-us, en 4, 28 -- MIME-Version: 1.0 4, 28 -- To: Microscopy-at-microscopy.com 4, 28 -- Subject: RE: Vibratome sections 4, 28 -- X-OriginalArrivalTime: 13 Feb 2007 20:55:26.0320 (UTC) 4, 28 -- FILETIME=[496F8700:01C74FB1] 4, 28 -- X-WSS-ID: 69CCF7A32AW570148-08-01 4, 28 -- Content-Type: text/plain; 4, 28 -- charset=iso-8859-1; 4, 28 -- format=flowed 4, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Vitaly, I don't think I've actually cleaned anything smaller than about 50u, but I also don't see any particular reason why 1u diamond shouldn't work on a 10u aperture. Give it a try.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Friday, February 09, 2007 12:05 AM To: kenconverse-at-qualityimages.biz
Jim, True, but does it affect the imaging? My experience with SEMs says no. I'm not sure about TEMs. Ken
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Sent: Wednesday, February 14, 2007 11:47 AM To: kenconverse-at-qualityimages.biz
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Question: We have a JEOL JSM 5310 LV scanning microscope (low vacuum, environmental type) with an Oxford Instruments EDS system and the LINK ISIS 300 software. I am trying to use SEM Quant for full quantitative analysis of minerals using a set of well characterized standards. I have done this before on a regular SEM very successfully, with the routine involving setting the working distance at 39 mm. However, with this LV SEM model, I have noticed that I am getting a very weak signal (very low number of counts) on my calibration standard (I'm using cobalt), but that the signal is appropriate if the working distance is reduced to 26 mm. My question therefore is:
How critical is it to stick with a working distance of 39 mm for quatitative analysis and subsequent ZAF corrections? If I change the WD, will I need to make any changes to my protocol for quantitative analysis? If that WD is very critical, is there a way to improve the count rate while still working at 39 mm?
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Email: tjj-at-stowers-institute.org Name: Teri Johnson
Organization: Stowers Institute for Medical Research
Title-Subject: [Filtered] Position available: Research EM Specialist
Question: Electron Microscopy Specialist The Stowers Institute for Medical Research has an opening for an Electron Microscopy (EM) Specialist to oversee day-to-day operations of the new electron microscopy service in the Histology Core Facility.
Responsibilities include providing high quality research EM services on biological samples using established protocols; daily operation of the electron microscopy lab; using and maintaining the microscopes and ancillary equipment; assisting researchers with specimen preparation; sample preparation, including sample receipt, fixation, processing, and embedding; development of protocols; training SIMR research members and other users of techniques and equipment; development of seminars and/or presentations for internal training; purchasing supplies; and maintaining records/archives of the EM lab operation.
In addition to excellent organizational, communication, and problem solving skills, the successful candidate should be familiar with operation of all applicable specimen preparation equipment and microscopes (TEM and SEM); skilled at standard specimen preparation protocols and identifying associated artifacts; have experience in high pressure freezing, cryoEM techniques, and immuno EM; be able to lift in excess of 30 pounds; and be able work overtime, including weekday, weekend, or on call work.
The minimum requirements include an Associates Degree in a biological science and four years experience in research EM. Or a four year degree in a biological science and two years experience in research EM.
The Institute is looking for talented and intelligent people of high integrity who exemplify the values of the Institute--that is, those who are sincere and dependable and who strive for excellence in all they do. We provide a work environment that respects the dignity of every individual and encourages collaboration, teamwork, creativity and innovation. Our founders, Jim and Virginia Stowers, envision creating one of the most innovative and effective medical research organizations in the world. This vision can only be achieved if we attract the very best people.
The Institute provides a positive and nondiscriminatory atmosphere for all applicants, members, and others participating in Institute programs. It is the policy of the Institute to afford equal opportunity in all phases of employment (including advertising, solicitation, recruitment, hiring, transfers, promotions, demotions, compensation, training, benefits, layoffs, terminations and all other terms and conditions of employment) to all individuals regardless of race, creed, color, religion, gender, sexual orientation, pregnancy, national origin, age, disability (including within the meaning of section 504 of the Rehabilitation Act), military status or any other status protected by law.
To apply, visit: http://www.stowers-institute.org/ScientistsSought/ScientistsSought.asp and scroll to the entry for the EM Specialist. Click on the "Apply Now" hyperlink.
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Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers Institute
Title-Subject: [Filtered] Leica EM-Stain
Question: Hello, I am interested in purchasing the Leica EM-Stain. Does anyone have any comments, positive and negative, that I should take into consideration before making such an investment? We will be using it on formvar coated slot grids. Thanks in advance, Rhonda Allen BA HT(ASCP)HTL, QIHC Stowers Institute for Medical Research Kansas City, Missouri 816-926-4346
First of all although the instrumant that you used in the past was optimised for EDS at 39mm most modern instruments work with a far shorter WD.
A simple way to determine the best WD is to place a stub in the instrument at a set magnification. Look st and the count rate. Move the specimen to a new position at the same magnification and refocus before checking the new count rate. Repeat until the optimum maximum count position is determined.
You will need to feed the new take off angle and azimuth (if any) into the software so that the corrections apply to the new postion.
} From my notes on the operation of the 5300 I see a working distance of 15mm seems to be ideal!
Good luck
Steve Chapman Protrain for EM training & consultancy world wide www.emcourses.com Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967
----- Original Message ----- X-from: {elshazly-at-marshall.edu} To: {protrain-at-emcourses.com} Sent: Wednesday, February 14, 2007 9:25 PM
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Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers Institute
Title-Subject: [Filtered] Leica EM Stain Disregard Please
Question: Please disregard the email from Rhonda Allen. I was trying to post something else, and that old question I asked last year keeps coming up.
Cheers,
Rhonda Allen Stowers Institute Kansas City, Missouri
I apologise as my last reply was in haste written between lectures (I am running a series of courses in Australia) I can see I did not explain clearly - try this
First of all although the instrument that you used in the past was optimised for EDS at 39mm most modern instruments work with a far shorter WD.
A simple way to determine the best WD is to place a stub in the instrument at a set magnification. Look at and record the count rate. Move the specimen Z to a new position at the same magnification and refocus before checking the new count rate. Repeat until the optimum, maximum count, Z position is determined, note the focal current or working distance for future reference. When you carry out an analysis, once this focal current or WD has been set, only adjust focus from area to area through a change in sample height in the stage - "Z".
You will need to feed the new take off angle and azimuth (if any) into the software so that the corrections apply to the new position.
} From my notes on the operation of the 5300 with clients I see a working distance of 15mm seems to be ideal!
Good luck, sorry my first reply was not complete.
----- Original Message ----- X-from: {elshazly-at-marshall.edu} To: {protrain-at-emcourses.com} Sent: Wednesday, February 14, 2007 9:25 PM
Hi Vitaly,
I have used diamond paste to clean both SEM and TEM apertures in emergencies.
Standard W filament SEMs have been fine and I have got away with it in a LaB6 filament, 200kV TEM, 50 um objective aperture (max mag 330K).
BUT I would not use it to clean apertures in a High Resolution TEM objective, neither would I use it to clean condenser apertures in a FEG or for small probe modes (apertures of 20um or less). These are quite unforgiving and need good apertures, I would not want to break the vacuum again for the sake of buying a new aperture or cleaning one properly. 'Properly' is the subject of other discussions.
Regards, Ron
In message {200702141802.l1EI2Tnu025620-at-ns.microscopy.com} kenconverse-at-qualityimages.biz writes: } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Jim, } True, but does it affect the imaging? My experience with SEMs says no. I'm } not sure about TEMs. } Ken } } QUALITY IMAGES } Servicing Scanning Electron Microscopes } Since 1981 } 474 So. Bridgton Rd. } Bridgton, ME 04009 } 207-647-4348 } Fax 207-647-2688 } kenconverse-at-qualityimages.biz } qualityimages.biz } } } } -----Original Message----- } X-from: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] } Sent: Wednesday, February 14, 2007 11:47 AM } To: kenconverse-at-qualityimages.biz } Subject: Re: [Microscopy] cleaning apertures chemically? } } } Ken } } 1um abrasive will leave 0.3 to 0.1 scratches. } These will be visible at the edges. } } Jim } } } } From mail-at-ns.microscopy.com Wed Feb 14 10:04:45 2007 } } Date: Wed, 14 Feb 2007 09:11:12 -0600 } } To: jquinn-at-www.matscieng.sunysb.edu } } From: kenconverse-at-qualityimages.biz } } Reply-to: kenconverse-at-qualityimages.biz } } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } } Subject: [Microscopy] cleaning apertures chemically? } } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } } X-lewp: MicroscopyListSpam NAGS } } } } } } } } } } } ---------------------------------------------------------------------------- } } 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 } } } ---------------------------------------------------------------------------- } } } } Vitaly, } } I don't think I've actually cleaned anything smaller than about 50u, but } I } also don't see any particular reason why 1u diamond shouldn't work on a } 10u } aperture. Give it a try. } } Ken Converse } owner } } QUALITY } IMAGES } Servicing Scanning Electron Microscopes } Since 1981 } 474 So. } Bridgton Rd. } Bridgton, ME 04009 } 207-647-4348 } Fax 207-647-2688 } } kenconverse-at-qualityimages.biz } qualityimages.biz } } } } -----Original } Message----- } X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] } } Sent: Friday, February 09, 2007 12:05 AM } } To: kenconverse-at-qualityimages.biz } } Subject: Re: [Microscopy] RE: cleaning apertures chemically? } } } } neat stuff. will it work for very small Pt and Mo apertures - say 10um obj. } } aperture in TEM? Sounds like a big time saver. } } } } Vitaly Feingold } } SIA } } 2773 Heath Lane } } Duluth GA 30096 } } Ph. 770-232-7785 } } Fax 770-232-1791 } } www.sia-cam.com } } ----- Original Message ----- } } X-from: {kenconverse-at-qualityimages.biz} } } To: {vitalylazar-at-att.net} } } Sent: Wednesday, February 07, 2007 10:05 PM } } Subject: [Microscopy] RE: cleaning apertures chemically? } } } } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } ------ } } } 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 } } } } } } ---------------------------------------------------------------------------- } } } } } } John, } } } I've had very good luck for the last 30 years or so cleaning apertures } } } with } } a cut knap polishing cloth and 1 micron diamond paste. Just } place the } } aperture on the cloth with a little paste and put your finger } on it and } } } rub } } } it in a circular motion. Do both sides. Clean ultrasonically in Joy } } } dishwashing liquid and hot water, rinse in hot tap water or distilled } } } water } } } and immediately blow dry with a duster to avoid water spots. My } } } understanding about Joy is that the Proctor & Gamble labs use it for } } } cleaning critical parts of AAUs and can find no residue. Apparently this } } } is } } } not true of all dishwashing liquids. } } } } } } This works with 1 mil foil (including multi-hole strips), 5 mil } } } countersunk } } and even the little Siemems apertures that are heavily } countersunk. Just } } don't try it with gold foil self-cleaning apertures. } The gold foil is far } } too thin and fragile for this technique. } } } } } Chuck Garber at SPI tells me that most metallographic diamond pastes } } } have silicones in them. That's a problem, but his pastes don't. As a } } } bonus, the SPI prices are very good. No financial interest, just a } } } happy user. } } } } } } As you are probably aware, heating moly in a platinum boat is a } } } problem } } } and } } } even heating Pt has limited usefulness because the Pt recrystalizes and } } } eventually you have an aperture with alligator skin and it won't work } any } } more (high astigmatism). Polishing a ruined Pt aperture will } restore it, } } probably due to the smearing of the metal by the diamond } particles. } } } } All in all it's pretty good because you don't need to } know what the } } aperture } } is made of, there's no complicated or } exotic equipment needed (beyond an } } ultrasonic cleaner), no organic } solvents or other nasty stuff, and you can } } use the same apertures for } years. Once in a great while I might fold a 1 } } mil aperture, but that } doesn't happen very often. } } } } Ken Converse } } Owner } } } } } QUALITY IMAGES } } Servicing Scanning Electron Microscopes } } Since 1981 } } } 474 So. Bridgton Rd. } } Bridgton, ME 04009 } } 207-647-4348 } } Fax } 207-647-2688 } } kenconverse-at-qualityimages.biz } } qualityimages.biz } } } } } } } } } -----Original Message----- } } X-from: johnf-at-geology.wisc.edu } [mailto:johnf-at-geology.wisc.edu] } } Sent: Wednesday, February 07, 2007 4:11 } PM } } To: kenconverse-at-qualityimages.biz } } Subject: [Microscopy] } cleaning apertures chemically? } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } ------ } } } 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 have any experience cleaning the carbonaeous gunk off of } } } Mo or Pt-Ir apertures with a solvent? I know about heating up in a Mo } } } etc boat, but looking for alternatives. Will ammonium hydroxide do the } } } trick? Or something else? } } } } } } thanks. } } } -- } } } ======================================================== } } } John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) } 438-7480 } } } Cameron Electron Microprobe Lab lab: (608) 265-4798 } } } Dept of Geology & Geophysics fax: (608) 262-0693 } } } University of Wisconsin home: (608) 274-2245 } } } 1215 West Dayton St. email: johnf-at-geology.wisc.edu } } } Madison, WI 53706 amateur radio: WA3BTA } } } Personal http://www.geology.wisc.edu/~johnf/ } } } Probe lab http://www.geology.wisc.edu/~johnf/sx51.html } } } Probe Sign Up Calender: } } } http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi } } } } } } "The first rule of all intelligent tinkering is to save every cog and } } } wheel." -- Aldo Leopold } } } } } } "For a successful technology, reality must take precedence over } } } public relations, for Nature cannot be fooled." -- Richard P. } } } Feynman } } } } } } ==============================Original } } } Headers============================== } } } 4, 24 -- From johnf-at-geology.wisc.edu Wed Feb 7 15:07:22 2007 4, 24 -- } } } Received: from ice.geology.wisc.edu (ice.geology.wisc.edu } [144.92.206.14]) } } 4, 24 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } } l17L7MvG015890 } } 4, 24 -- } for {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:22 } } -0600 } } 4, } 24 -- Received: from localhost (localhost [127.0.0.1]) } } 4, 24 -- by } localhost (Postfix) with ESMTP id 9A3E520D20 } } 4, 24 -- for } {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:21 } } -0600 (CST) } } } 4, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) } } 4, 24 -- by } localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port } } 10024) 4, } 24 -- with ESMTP id 07529-02 for {microscopy-at-microscopy.com} ; } } } 4, } } } 24 -- Wed, 7 Feb 2007 15:07:14 -0600 (CST) 4, 24 -- Received: from } } } [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) } } 4, 24 -- } (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) } } 4, 24 -- } (No client certificate requested) } } 4, 24 -- by ice.geology.wisc.edu } (Postfix) with ESMTP id 29B0A20D02 } } 4, 24 -- for } {microscopy-at-microscopy.com} ; Wed, 7 Feb 2007 15:07:14 } } -0600 (CST) } } } 4, 24 -- Mime-Version: 1.0 } } 4, 24 -- Message-Id: } {p06230910c1efee78374f-at-[144.92.206.57]} } } } 4, 24 -- Date: Wed, 7 Feb 2007 15:03:05 -0600 } } } 4, 24 -- To: microscopy-at-microscopy.com } } } 4, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} } } } 4, 24 -- Subject: cleaning apertures chemically? } } } 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 4, } } 24 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu } } } ==============================End of - } } } Headers============================== } } } } } } } } } } } } } } } } } } _________________________________________________________________ } } } Need personalized email and website? 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Learn more at www.doteasy.com } } } } } } } } } ==============================Original } } } Headers============================== } } } 25, 29 -- From kenconverse-at-qualityimages.biz Wed Feb 7 21:03:14 2007 } } } 25, 29 -- Received: from dpmailmta02.doteasy.com } (dpmailmta02.doteasy.com } } } [65.61.209.81]) } } } 25, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } } l1833EcY005037 } } } 25, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Feb 2007 } } } 21:03:14 -0600 } } } 25, 29 -- Received: from qualityimages.biz (unverified } [192.168.101.16]) } } 25, 29 -- by dpmailmta02.doteasy.com (DEO) with ESMTP } id 57086489-1814644 } } 25, 29 -- for multiple; Wed, 07 Feb 2007 19:43:42 } -0800 } } 25, 29 -- Received: from Ken [68.193.55.199] by qualityimages.biz } with } } } ESMTP } } } 25, 29 -- (SMTPD32-8.05) id A2EEE2DE004C; Wed, 07 Feb 2007 } } } 19:03:10 -0800 } } } 25, 29 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} } } 25, } 29 -- To: {johnf-at-geology.wisc.edu} } } 25, 29 -- Cc: "MSA Listserver" } {Microscopy-at-MSA.Microscopy.Com} } } 25, 29 -- Subject: RE: [Microscopy] } cleaning apertures chemically? } } 25, 29 -- Date: Wed, 7 Feb 2007 22:02:53 } -0500 } } 25, 29 -- Message-ID: {006501c74b2d$a21cb3b0$8d7ec44b-at-Ken} } } } 25, 29 -- MIME-Version: 1.0 } } } 25, 29 -- Content-Type: text/plain; } } } 25, 29 -- charset="us-ascii" } } } 25, 29 -- X-Priority: 3 (Normal) } } } 25, 29 -- X-MSMail-Priority: Normal } } } 25, 29 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 } } } 25, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } } } 25, 29 -- In-Reply-To: {200702072111.l17LB5uK020615-at-ns.microscopy.com} } } } 25, 29 -- Importance: Normal } } } 25, 29 -- X-IMSTrailer: __IMail_7__ } } } 25, 29 -- X-IP-stats: Incoming Last 0, First 133, in=3174090, out=0, } } } spam=0 ip=192.168.101.16 } } } 25, 29 -- X-External-IP: 192.168.101.16 } } } 25, 29 -- Content-Transfer-Encoding: 8bit } } } 25, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } } ns.microscopy.com id l1833EcY005037 } } } ==============================End of - } } } Headers============================== } } } } } } } } } } } } } } _________________________________________________________________ } } Need personalized email and website? 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Learn more at www.doteasy.com } } } ==============================Original Headers============================== } 17, 30 -- From kenconverse-at-qualityimages.biz Wed Feb 14 11:54:57 2007 } 17, 30 -- Received: from dpmailmta02.doteasy.com (dpmailmta02.doteasy.com [65.61.209.85]) } 17, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1EHsueB014851 } 17, 30 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 14 Feb 2007 11:54:56 - 0600 } 17, 30 -- Received: from qualityimages.biz (unverified [192.168.101.16]) } 17, 30 -- by dpmailmta02.doteasy.com (DEO) with ESMTP id 59614064-1814644 } 17, 30 -- for multiple; Wed, 14 Feb 2007 10:38:07 -0800 } 17, 30 -- Received: from Ken [76.179.237.214] by qualityimages.biz with ESMTP } 17, 30 -- (SMTPD32-8.05) id AC5D245F00EC; Wed, 14 Feb 2007 09:52:29 -0800 } 17, 30 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} } 17, 30 -- To: "'Jim Quinn'" {jquinn-at-www.matscieng.sunysb.edu} } 17, 30 -- Cc: "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} } 17, 30 -- Subject: RE: [Microscopy] cleaning apertures chemically? } 17, 30 -- Date: Wed, 14 Feb 2007 12:54:16 -0500 } 17, 30 -- Message-ID: {000301c75061$279f4e70$6401a8c0-at-Ken} } 17, 30 -- MIME-Version: 1.0 } 17, 30 -- Content-Type: text/plain; } 17, 30 -- charset="us-ascii" } 17, 30 -- X-Priority: 3 (Normal) } 17, 30 -- X-MSMail-Priority: Normal } 17, 30 -- X-Mailer: Microsoft Outlook, Build 10.0.6822 } 17, 30 -- Thread-Index: AcdQWFb7TWkd2t1SS8i/qbSh58h5dwACIRYg } 17, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } 17, 30 -- Importance: Normal } 17, 30 -- In-Reply-To: {200702141646.l1EGkWT05777-at-www.matscieng.sunysb.edu} } 17, 30 -- X-IMSTrailer: __IMail_7__ } 17, 30 -- X-IP-stats: Incoming Last 0, First 139, in=3299568, out=0, spam=0 ip=192.168.101.16 } 17, 30 -- X-Originating-IP: 192.168.101.16 } 17, 30 -- Content-Transfer-Encoding: 8bit } 17, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1EHsueB014851 } ==============================End of - Headers============================== }
-- Mr. Ron Doole Department of Materials Senior Instrumentation Engineer. University of Oxford. Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
==============================Original Headers============================== 8, 24 -- From ron.doole-at-materials.ox.ac.uk Thu Feb 15 02:25:45 2007 8, 24 -- Received: from relay6.mail.ox.ac.uk (relay6.mail.ox.ac.uk [163.1.2.167]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1F8Pi1s011484 8, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Feb 2007 02:25:45 -0600 8, 24 -- Received: from webmail220.herald.ox.ac.uk ([163.1.0.220]) 8, 24 -- by relay6.mail.ox.ac.uk with esmtp (Exim 4.62) 8, 24 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 8, 24 -- id 1HHbw0-0004nz-KC 8, 24 -- for Microscopy-at-microscopy.com; Thu, 15 Feb 2007 08:25:44 +0000 8, 24 -- Received: by webmail220.herald.ox.ac.uk (Postfix, from userid 33) 8, 24 -- id 25C6394047; Thu, 15 Feb 2007 08:25:44 +0000 (GMT) 8, 24 -- Content-Type: text/plain 8, 24 -- Content-Disposition: inline 8, 24 -- Content-Transfer-Encoding: 7bit 8, 24 -- MIME-Version: 1.0 8, 24 -- X-Mailer: MIME-tools 5.417 (Entity 5.417) 8, 24 -- From: Ron Doole {ron.doole-at-materials.ox.ac.uk} 8, 24 -- Date: Thu, 15 Feb 2007 08:25:44 +0000 8, 24 -- To: Microscopy-at-microscopy.com 8, 24 -- Subject: Re: [Microscopy] RE: cleaning apertures chemically? 8, 24 -- In-Reply-To: {200702141802.l1EI2Tnu025620-at-ns.microscopy.com} 8, 24 -- X-Webmail-Sender: rdoole 8, 24 -- X-Webmail-Originating-Ip: 129.67.84.180 8, 24 -- Message-Id: {20070215082544.25C6394047-at-webmail220.herald.ox.ac.uk} ==============================End of - Headers==============================
After opening the ISIS application select the dewar icon, then 'detector' then 'orientation'. This should list the detector conditions including the working distance.
Ron
} } Title-Subject: [Filtered] Takeoff angle } } Question: We have a JEOL JSM 5310 LV scanning microscope (low vacuum, } environmental type) with an Oxford Instruments EDS system and the } LINK ISIS 300 software. I am trying to use SEM Quant for full } quantitative analysis of minerals using a set of well characterized } standards. I have done this before on a regular SEM very } successfully, with the routine involving setting the working distance } at 39 mm. However, with this LV SEM model, I have noticed that I am } getting a very weak signal (very low number of counts) on my } calibration standard (I'm using cobalt), but that the signal is } appropriate if the working distance is reduced to 26 mm. My question } therefore is: } } How critical is it to stick with a working distance of 39 mm for } quatitative analysis and subsequent ZAF corrections? If I change the } WD, will I need to make any changes to my protocol for quantitative } analysis? } If that WD is very critical, is there a way to improve the count rate } while still working at 39 mm? } } Thank you for your cooperation. } } Aley } } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 10, 12 -- From zaluzec-at-microscopy.com Wed Feb 14 15:22:02 2007 } 10, 12 -- Received: from [192.168.1.27] (msdvpn8.msd.anl.gov [130.202.238.72]) } 10, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1ELLxm8030451 } 10, 12 -- for {microscopy-at-microscopy.com} ; Wed, 14 Feb 2007 15:22:00 -0600 } 10, 12 -- Mime-Version: 1.0 } 10, 12 -- X-Sender: (Unverified) } 10, 12 -- Message-Id: {p06020405c1f92f904e4b-at-[192.168.1.27]} } 10, 12 -- Date: Thu, 15 Feb 2007 08:29:01 +1100 } 10, 12 -- To: microscopy-at-microscopy.com } 10, 12 -- From: elshazly-at-marshall.edu (by way of MicroscopyListserver) } 10, 12 -- Subject: viaWWW: Takeoff angle } 10, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers============================== }
-- Mr. Ron Doole Department of Materials Senior Instrumentation Engineer. University of Oxford. Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
==============================Original Headers============================== 6, 24 -- From ron.doole-at-materials.ox.ac.uk Thu Feb 15 02:51:20 2007 6, 24 -- Received: from relay2.mail.ox.ac.uk (relay2.mail.ox.ac.uk [163.1.2.161]) 6, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1F8pJwl022963 6, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Feb 2007 02:51:20 -0600 6, 24 -- Received: from webmail220.herald.ox.ac.uk ([163.1.0.220]) 6, 24 -- by relay2.mail.ox.ac.uk with esmtp (Exim 4.62) 6, 24 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 6, 24 -- id 1HHcKl-0004H4-7H; Thu, 15 Feb 2007 08:51:19 +0000 6, 24 -- Received: by webmail220.herald.ox.ac.uk (Postfix, from userid 33) 6, 24 -- id 2165694047; Thu, 15 Feb 2007 08:51:19 +0000 (GMT) 6, 24 -- Content-Type: text/plain 6, 24 -- Content-Disposition: inline 6, 24 -- Content-Transfer-Encoding: 7bit 6, 24 -- MIME-Version: 1.0 6, 24 -- X-Mailer: MIME-tools 5.417 (Entity 5.417) 6, 24 -- From: Ron Doole {ron.doole-at-materials.ox.ac.uk} 6, 24 -- Date: Thu, 15 Feb 2007 08:51:19 +0000 6, 24 -- To: elshazly-at-marshall.edu 6, 24 -- CC: Microscopy-at-microscopy.com 6, 24 -- Subject: Re: [Microscopy] viaWWW: Takeoff angle 6, 24 -- In-Reply-To: {200702142133.l1ELXjhx030086-at-ns.microscopy.com} 6, 24 -- X-Webmail-Sender: rdoole 6, 24 -- X-Webmail-Originating-Ip: 129.67.84.180 6, 24 -- Message-Id: {20070215085119.2165694047-at-webmail220.herald.ox.ac.uk} ==============================End of - Headers==============================
Hello Everybody, I would like to address the collective wisdom on neutral density filters. I'm using Hoya camera ND filters to lower light levels for photomicroscopy while maintaining color temperature. But I've convinced myself that these filters have a slight greenish cast. Is this just my fancy running away with me?
Who makes a good grade of ND filter. I don't want to use the Kodak gel filters as I prefer the weight and strength of glass. Any suggestions?
Stay safe............ Frank
==============================Original Headers============================== 5, 17 -- From frank.karl-at-degussa.com Thu Feb 15 09:49:49 2007 5, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1FFnmk0014302 5, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 15 Feb 2007 09:49:49 -0600 5, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 5, 17 -- by malmailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with SMTP id l1FFneJB017572 5, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 15 Feb 2007 16:49:46 +0100 5, 17 -- Subject: ND filters 5, 17 -- To: microscopy-at-msa.microscopy.com 5, 17 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 5, 17 -- Message-ID: {OF36B70ADD.F777CD4F-ON86257283.005651D9-85257283.0056ECF2-at-degussa.com} 5, 17 -- From: frank.karl-at-degussa.com 5, 17 -- Date: Thu, 15 Feb 2007 10:49:28 -0500 5, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 5, 17 -- 02/15/2007 09:49:47 AM 5, 17 -- MIME-Version: 1.0 5, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
If they are true ND filters made correctly by a reliable manufacturer there will be no greenish cast. Tiffen, Hoya, Nikon, etc., should all be free of color casts. Can't vouch for "Acme Filter Co." or "Joe's Filters".
That said, there may be issues of older filters having developed an off-color as dyes or plastics age. I'm not extremely familiar with the various methods of making filters, but I strongly suspect that some methods are more stable than others over geological time.
Cheers, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: Thursday, February 15, 2007 9:55 AM To: Tindall, Randy D.
Hello Everybody, I would like to address the collective wisdom on neutral density filters. I'm using Hoya camera ND filters to lower light levels for photomicroscopy while maintaining color temperature. But I've convinced myself that these filters have a slight greenish cast. Is this just my fancy running away with me?
Who makes a good grade of ND filter. I don't want to use the Kodak gel filters as I prefer the weight and strength of glass. Any suggestions?
Stay safe............ Frank
==============================Original Headers============================== 5, 17 -- From frank.karl-at-degussa.com Thu Feb 15 09:49:49 2007 5, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1FFnmk0014302 5, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 15 Feb 2007 09:49:49 -0600 5, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 5, 17 -- by malmailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with SMTP id l1FFneJB017572 5, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 15 Feb 2007 16:49:46 +0100 5, 17 -- Subject: ND filters 5, 17 -- To: microscopy-at-msa.microscopy.com 5, 17 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 5, 17 -- Message-ID: {OF36B70ADD.F777CD4F-ON86257283.005651D9-85257283.0056ECF2-at-degussa.com} 5, 17 -- From: frank.karl-at-degussa.com 5, 17 -- Date: Thu, 15 Feb 2007 10:49:28 -0500 5, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 5, 17 -- 02/15/2007 09:49:47 AM 5, 17 -- MIME-Version: 1.0 5, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 16, 26 -- From TindallR-at-missouri.edu Thu Feb 15 10:02:06 2007 16, 26 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 16, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1FG26Qc023693 16, 26 -- for {microscopy-at-microscopy.com} ; Thu, 15 Feb 2007 10:02:06 -0600 16, 26 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 16, 26 -- Thu, 15 Feb 2007 10:02:06 -0600 16, 26 -- x-mimeole: Produced By Microsoft Exchange V6.5 16, 26 -- Content-class: urn:content-classes:message 16, 26 -- MIME-Version: 1.0 16, 26 -- Content-Type: text/plain; 16, 26 -- charset="us-ascii" 16, 26 -- Subject: RE: [Microscopy] ND filters 16, 26 -- Date: Thu, 15 Feb 2007 10:02:06 -0600 16, 26 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68E10-at-UM-XMAIL08.um.umsystem.edu} 16, 26 -- In-Reply-To: {200702151554.l1FFsdlx016336-at-ns.microscopy.com} 16, 26 -- X-MS-Has-Attach: 16, 26 -- X-MS-TNEF-Correlator: 16, 26 -- Thread-Topic: [Microscopy] ND filters 16, 26 -- Thread-Index: AcdRGZn7kRFw3ICtQGKJxQIdUp04RgAAGDwg 16, 26 -- References: {200702151554.l1FFsdlx016336-at-ns.microscopy.com} 16, 26 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 16, 26 -- To: {frank.karl-at-degussa.com} 16, 26 -- Cc: {microscopy-at-microscopy.com} 16, 26 -- X-OriginalArrivalTime: 15 Feb 2007 16:02:06.0199 (UTC) FILETIME=[A3C5B070:01C7511A] 16, 26 -- Content-Transfer-Encoding: 8bit 16, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1FG26Qc023693 ==============================End of - Headers==============================
Other replies have pointed out that the geometry for analysis varies between microscopes. We are still running a JEOL 840A that is setup for EDX at a working distance of 39mm. Our Hitachi 2460N uses 25mm, and we are hoping to get a new SEM that uses something between 5 and 10mm.
If your ISIS system was installed by the Oxford personnel then the proper geometric constants, including working distance, should have already been set up. You should have a Detector icon on the Labbook task manager. Start it, and then choose the Detector menu and the Orientation option. That should show you the preferred working distance for your microscope.
You probably still want to perform the exercise of confirming the working distance for your maximum count rate. However, if the collimator on your x-ray detector has a cut out on the bottom, it is possible that you will get an optimum count rate at some slightly greater working distance. That would not be good as you want to be at the specified working distance. I have also found that indicated working distance is not the same for all voltages. I can set our Hitachi objective lens to a nominal 25mm focal length and find that I have to raise or lower the stage as I go to different accelerating voltages to bring the image back into focus. You will need to determine the proper indicated working distance at a given kV for your actual desired working distance.
One other thing - you need to make sure that the parameters stored with your spectra reflect your actual conditions and geometry. MANY years ago, I collected several weeks of data on a Kevex system before I realized that we had not setup the software to store the correct conditions. Fortunately, we were consistent in our data collection and were able to correct the files, but it is better to get it right the first time.
We have an Oxford ISIS on our Hitachi and the provision for reading conditions directly from the microscope. However, we also have a utility for suspending that communication for certain operations. In that situation, the last known conditions get stored with the spectra. If someone suspends communication while the beam is turned off, an accelerating voltage of zero is the last known value. That plays even more havoc with ZAF corrections than does a wrong take-off angle.
EDS can produce some pretty decent results, but a good amount of care in setup and collection is necessary. Just because results are easily produced with the press of a button doesn't mean they are right.
Warren Straszheim
-----Original Message----- Sent: Wednesday, February 14, 2007 3:25 PM
Question: We have a JEOL JSM 5310 LV scanning microscope (low vacuum, environmental type) with an Oxford Instruments EDS system and the LINK ISIS 300 software. I am trying to use SEM Quant for full quantitative analysis of minerals using a set of well characterized standards. I have done this before on a regular SEM very successfully, with the routine involving setting the working distance at 39 mm. However, with this LV SEM model, I have noticed that I am getting a very weak signal (very low number of counts) on my calibration standard (I'm using cobalt), but that the signal is appropriate if the working distance is reduced to 26 mm. My question therefore is:
How critical is it to stick with a working distance of 39 mm for quatitative analysis and subsequent ZAF corrections? If I change the WD, will I need to make any changes to my protocol for quantitative analysis? If that WD is very critical, is there a way to improve the count rate while still working at 39 mm?
Thank you for your cooperation.
Aley
==============================Original Headers============================== 16, 32 -- From wesaia-at-iastate.edu Thu Feb 15 10:12:07 2007 16, 32 -- Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) 16, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1FGC6be004776 16, 32 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 15 Feb 2007 10:12:06 -0600 16, 32 -- Received: from mailout-1.iastate.edu (mailout-1.iastate.edu [129.186.140.1]) 16, 32 -- by mailhub-3.iastate.edu (8.12.11.20060614/8.12.10) with SMTP id l1FG8vMZ008720; 16, 32 -- Thu, 15 Feb 2007 10:08:57 -0600 16, 32 -- Received: from (owa.eng.iastate.edu [129.186.23.85]) by mailout-1.iastate.edu with smtp 16, 32 -- id 20fa_9456a4cc_bd0e_11db_8423_00137253420a; 16, 32 -- Thu, 15 Feb 2007 10:07:04 -0600 16, 32 -- Received: from maire.eng.iastate.edu ([10.10.196.69]) by owa.eng.iastate.edu with Microsoft SMTPSVC(6.0.3790.1830); 16, 32 -- Thu, 15 Feb 2007 10:08:57 -0600 16, 32 -- Content-class: urn:content-classes:message 16, 32 -- MIME-Version: 1.0 16, 32 -- Content-Type: text/plain; 16, 32 -- charset="us-ascii" 16, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 32 -- Subject: RE: [Microscopy] viaWWW: Takeoff angle 16, 32 -- Date: Thu, 15 Feb 2007 10:11:27 -0600 16, 32 -- Message-ID: {16A330AC32056A40B32842EC4BB8D72701629456-at-maire.eng.iastate.edu} 16, 32 -- In-Reply-To: {200702142124.l1ELOjge002029-at-ns.microscopy.com} 16, 32 -- X-MS-Has-Attach: 16, 32 -- X-MS-TNEF-Correlator: 16, 32 -- Thread-Topic: [Microscopy] viaWWW: Takeoff angle 16, 32 -- Thread-Index: AcdQfo1auAw5lE3QQwe7AZ8tFmAt8gAl/0cw 16, 32 -- References: {200702142124.l1ELOjge002029-at-ns.microscopy.com} 16, 32 -- From: "Straszheim, Warren E [CCE E]" {wesaia-at-iastate.edu} 16, 32 -- To: "MSA" {Microscopy-at-msa.microscopy.com} 16, 32 -- Cc: {elshazly-at-marshall.edu} 16, 32 -- X-OriginalArrivalTime: 15 Feb 2007 16:08:57.0525 (UTC) FILETIME=[98F10E50:01C7511B] 16, 32 -- Content-Transfer-Encoding: 8bit 16, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l1FGC6be004776 ==============================End of - Headers==============================
Frank, I agree with Randy Tindall that 'good quality' NDF's are nutral in the visible. But if yours has gone green for whatever reason, you can make nice ones yourslf by evaporating aluminum foil on glass. Of course, you have to be careful of the surface, but by changing the time of evap (and/or distance to the source) you can get nice NDFs pretty cheap (assuming you have an evaporator).
My two photons, Tobias
} ----------------------------------- } } Hello Everybody, } I would like to address the collective wisdom on neutral density filters. } I'm using Hoya camera ND filters to lower light levels for photomicroscopy } while maintaining color temperature. But I've convinced myself that these } filters have a slight greenish cast. Is this just my fancy running away } with me? } } Who makes a good grade of ND filter. I don't want to use the Kodak gel } filters as I prefer the weight and strength of glass. Any suggestions? } } Stay safe............ Frank }
Assuming we are talking about digital cameras, there is a wonderful little freeware program that will allow you to correct for problems with illumination color, uneven illumination, and the color response of your camera sensor. The program is "Image Arithmetic" and can be downloaded from http://www.t3i.nl/myblog/?page_id=7. The feature of interest is image division. The subject image is divided, pixel by pixel, by a blank image taken under identical conditions of illumination. You must do this in manual mode to keep the illumination intensity identical. Obviously the extra steps will slow things down, but the procedure works very well when done correctly.
Ralph Common Dept. of Physiology Michigan State University
==============================Original Headers============================== 3, 24 -- From rcommon-at-msu.edu Thu Feb 15 10:38:55 2007 3, 24 -- Received: from sys29.mail.msu.edu (sys29.mail.msu.edu [35.9.75.129]) 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1FGctYr027665 3, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Feb 2007 10:38:55 -0600 3, 24 -- Received: from [35.9.122.125] (helo=emlab) 3, 24 -- by sys29.mail.msu.edu with esmtpsa (Exim 4.52 #1) 3, 24 -- (TLSv1:RC4-MD5:128) 3, 24 -- id 1HHjdG-0006Ia-Vm 3, 24 -- for Microscopy-at-microscopy.com; Thu, 15 Feb 2007 11:38:55 -0500 3, 24 -- From: "Ralph Common" {rcommon-at-msu.edu} 3, 24 -- To: {Microscopy-at-microscopy.com} 3, 24 -- Subject: ND filters 3, 24 -- Date: Thu, 15 Feb 2007 11:40:28 -0500 3, 24 -- Message-ID: {00a001c75120$00bf9f70$7d7a0923-at-msu.edu} 3, 24 -- MIME-Version: 1.0 3, 24 -- Content-Type: text/plain; 3, 24 -- charset="iso-8859-1" 3, 24 -- Content-Transfer-Encoding: 7bit 3, 24 -- X-Priority: 3 (Normal) 3, 24 -- X-MSMail-Priority: Normal 3, 24 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) 3, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1896 3, 24 -- Importance: Normal 3, 24 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
I have a situation with an image analysis system attached to an optical metallograph that I would appreciate comments on. We are evaluating thermal spay coating for area percent porosity. Our program has both single phase and multi-phase area percent analyses sub-routines. Keeping the region of interest (field) constant, and the discrimination threshold also constant, the two phase analysis programs return different values (eg 3.2% vs 5.5%). Multiple operators have experienced the same issue.
Thanks in advance!
Chris Holp FirstEnergy Corp. Beta Labs Mayfield Village, OH 44143
440-604-9704
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==============================Original Headers============================== 7, 20 -- From holpc-at-firstenergycorp.com Thu Feb 15 11:05:03 2007 7, 20 -- Received: from firstenergycorp.com (gw09.firstenergycorp.com [205.132.74.166]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l1FH53iN006933 7, 20 -- for {microscopy-at-microscopy.com} ; Thu, 15 Feb 2007 11:05:03 -0600 7, 20 -- Subject: Image analysis 7, 20 -- To: Microscopy-at-microscopy.com 7, 20 -- X-Mailer: Lotus Notes Release 6.5.4 CCH5 September 12, 2005 7, 20 -- Message-ID: {OF1D61D900.EEA0B243-ON85257283.0059D23C-85257283.005B6242-at-FirstEnergyCorp.com} 7, 20 -- From: holpc-at-firstenergycorp.com 7, 20 -- Date: Thu, 15 Feb 2007 11:38:09 -0500 7, 20 -- MIME-Version: 1.0 7, 20 -- X-MIMETrack: Serialize by Router on mail01/Servers/FirstEnergy(Release 6.55FP1HF75 | July 7, 20 -- 21, 2006) at 02/15/2007 11:38:09, 7, 20 -- Itemize by SMTP Server on GW11/Servers/FirstEnergy(Release 6.5.5FP1|April 7, 20 -- 11, 2006) at 02/15/2007 11:38:07 AM, 7, 20 -- Serialize by Router on GW11/Servers/FirstEnergy(Release 6.5.5FP1|April 11, 2006) at 7, 20 -- 02/15/2007 11:38:07 AM, 7, 20 -- Serialize complete at 02/15/2007 11:38:07 AM 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="US-ASCII" ==============================End of - Headers==============================
Offhand, it sounds like a thresholding issue. You should have some way to review the thresholded image to compare it to the original. Area measurements can be quite sensitive to the settings, and your results seem to indicate that your application is one of those sensitive cases. It does raise the question of confidence in the answer when the results vary so widely.
What are the differences between single and multiple phase modes of operation? Offhand, I think you would want single-phase mode, but multi-phase mode should collapse to single-phase mode and give you the same answer if the same threshold was used. If your system automatically chooses a threshold, I would be very wary of the results. I prefer operator setting and review to make sure you are measuring what you want to.
Warren
-----Original Message----- X-from: holpc-at-firstenergycorp.com [mailto:holpc-at-firstenergycorp.com] Sent: Thursday, February 15, 2007 11:06 AM To: wesaia-at-iastate.edu
Chris writes ...
} I have a situation with an image analysis system attached to } an optical metallograph that I would appreciate comments on. } We are evaluating thermal spay coating for area percent } porosity. Our program has both single phase and multi-phase } area percent analyses sub-routines. Keeping the region of } interest (field) constant, and the discrimination threshold } also constant, the two phase analysis programs return } different values (eg 3.2% vs 5.5%). } Multiple operators have experienced the same issue.
At first glance 3.2 relative to 5.5 is a huge difference, but digital image thresholding also implies these conjugate values, 96.8 and 94.5, which are not so different. The small difference between the 2 softwares might reflect their different philosophies regarding how to treat the pixels at the edge of the ROI and/or image frame. The thresholding may also have some differences. For example, one might include in the count only those pixel values greater than the threshold value, while the other includes what is greater OR equal to the threshold.
You may be able to figure out what is going on by downsizing the same image to fewer and fewer pixels. At some point I would believe both softwares would give you the same answer and thereby provide clues as to what's happenin'
HTH & good luck :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 7, 21 -- From michael-at-shaffer.net Thu Feb 15 11:45:39 2007 7, 21 -- Received: from n007.sc1.he.tucows.com (smtpout1430.sc1.he.tucows.com [64.97.157.130]) 7, 21 --