A colleague wants to prepare bundles of polyester fibres for SEM by embedding and cutting with a Microm HM325 microtome using a steel knife. The idea is to examine the cut surface of the block to study the cross-sectional shape of the fibres.
Could you please recommend the best type of embedding material for the purpose? It should not contain aggressive solvents, or be cured at too high a temperature.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
==============================Original Headers============================== 6, 21 -- From hinmeigeng-at-hotmail.com Sat Apr 1 04:30:21 2006 6, 21 -- Received: from hotmail.com (bay101-f34.bay101.hotmail.com [64.4.56.44]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k31AUKTF011498 6, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sat, 1 Apr 2006 04:30:21 -0600 6, 21 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 6, 21 -- Sat, 1 Apr 2006 02:30:20 -0800 6, 21 -- Message-ID: {BAY101-F34425505819B73D3958F3ECAD70-at-phx.gbl} 6, 21 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 6, 21 -- Sat, 01 Apr 2006 10:30:17 GMT 6, 21 -- X-Originating-IP: [86.128.212.193] 6, 21 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 6, 21 -- X-Sender: hinmeigeng-at-hotmail.com 6, 21 -- Reply-To: R.H.Olley-at-reading.ac.uk 6, 21 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 6, 21 -- To: Microscopy-at-MSA.Microscopy.Com 6, 21 -- Cc: R.H.Olley-at-reading.ac.uk 6, 21 -- Subject: Polyester Fibre Sectioning 6, 21 -- Date: Sat, 01 Apr 2006 10:30:17 +0000 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; format=flowed 6, 21 -- X-OriginalArrivalTime: 01 Apr 2006 10:30:20.0902 (UTC) FILETIME=[471A6860:01C65577] ==============================End of - Headers==============================
The 2006 Workshop run by Steve Chapman (Protrain Ltd, UK) will be held in Canberra at the Australian National University Electron Microscopy Unit.
The workshop will run over 5 days from the 12th to the 16th June 2006, and will cover SEM operation and basic maintenance, with some EDXA and FIB work.
The course in 2006 will be run in association with Anaspec cc Australia. The cost will be $AUS800 for the 5-day course, which includes morning and afternoon tea but no other meals or accommodation.
Contactfor details and bookings: Ruth O'Loughlin, Ruth-at-anaspec.co.za Canberra travel and accommodation suggestions can be found on
These highly recommended courses are enjoyable, intensive and interactive. Participants gain experience in getting the best possible performance from a range of SEMs. Professional microscopists and actual and aspiring "power users" are the main clientele, some people have so much fun they come back to do the course twice!
Accommodation: at ANU http://accom.anu.edu.au/ Other local accommodation : http://canberra.citysearch.com.au/section/visitor-guide/
Transport to Canberra - air, train, bus or road...http://canberra.citysearch.com.au/feature/51/gettingThere.html
Sally Stowe
-- Dr SJ Stowe ANU Electron Microscopy Unit http://www.anu.edu.au/EMU/index.html
==============================Original Headers============================== 11, 26 -- From sally.stowe-at-anu.edu.au Sat Apr 1 08:15:07 2006 11, 26 -- Received: from mail.rsbs.anu.edu.au (mail.rsbs.anu.edu.au [150.203.72.70]) 11, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k31EF6D2023777 11, 26 -- for {microscopy-at-microscopy.com} ; Sat, 1 Apr 2006 08:15:07 -0600 11, 26 -- Received: from mail.rsbs.anu.edu.au (mail.rsbs.anu.edu.au [127.0.0.1]) 11, 26 -- by mail.rsbs.anu.edu.au (Postfix) with SMTP 11, 26 -- id BD6AAF441E1; Sun, 2 Apr 2006 01:15:22 +1100 (EST) 11, 26 -- Received: from 203.113.234.139 11, 26 -- (SquirrelMail authenticated user u8411377) 11, 26 -- by mail.rsbs.anu.edu.au with HTTP; 11, 26 -- Sun, 2 Apr 2006 00:15:22 +1000 (EST) 11, 26 -- Message-ID: {1412.203.113.234.139.1143900922.squirrel-at-mail.rsbs.anu.edu.au} 11, 26 -- Date: Sun, 2 Apr 2006 00:15:22 +1000 (EST) 11, 26 -- Subject: Protrain SEM Workshop Canberra June 2006 11, 26 -- From: sally.stowe-at-anu.edu.au 11, 26 -- To: microscopy-at-microscopy.com, austem-at-anu.edu.au 11, 26 -- User-Agent: SquirrelMail/1.4.2-1 11, 26 -- MIME-Version: 1.0 11, 26 -- Content-Type: text/plain;charset=iso-8859-1 11, 26 -- Content-Transfer-Encoding: 8bit 11, 26 -- X-Priority: 3 11, 26 -- Importance: Normal 11, 26 -- X-RSBS-MailScanner-Information: Please contact the ISP for more information 11, 26 -- X-RSBS-MailScanner: Found to be clean 11, 26 -- X-RSBS-MailScanner-SpamScore: s 11, 26 -- X-MailScanner-From: sally.stowe-at-anu.edu.au ==============================End of - Headers==============================
We routinely observe fibre (all kinds - polymers, wools, cotton etc) cross sections in the sem, but rather than embedding them in resin, we feed the the bundle through the heat shrink tubing used in the electronics industry (as narrow as possible), shrink the tubing and then cut them using a sharp blade (we have found injector blades to be the best) on a Hardy microtome or similiar.
We have found this to be far less time consuming than embedding in a resin with very similar results.
Cheers
Colin Veitch CSIRO Textile and Fibre Technology Geelong, Australia
-----Original Message----- X-from: hinmeigeng-at-hotmail.com [mailto:hinmeigeng-at-hotmail.com] Sent: Sat 1/04/2006 9:30 PM To: Veitch, Colin (TFT, Geelong) Cc:
Greetings All !
A colleague wants to prepare bundles of polyester fibres for SEM by embedding and cutting with a Microm HM325 microtome using a steel knife. The idea is to examine the cut surface of the block to study the cross-sectional shape of the fibres.
Could you please recommend the best type of embedding material for the purpose? It should not contain aggressive solvents, or be cured at too high a temperature.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
==============================Original Headers============================== 6, 21 -- From hinmeigeng-at-hotmail.com Sat Apr 1 04:30:21 2006 6, 21 -- Received: from hotmail.com (bay101-f34.bay101.hotmail.com [64.4.56.44]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k31AUKTF011498 6, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sat, 1 Apr 2006 04:30:21 -0600 6, 21 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 6, 21 -- Sat, 1 Apr 2006 02:30:20 -0800 6, 21 -- Message-ID: {BAY101-F34425505819B73D3958F3ECAD70-at-phx.gbl} 6, 21 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 6, 21 -- Sat, 01 Apr 2006 10:30:17 GMT 6, 21 -- X-Originating-IP: [86.128.212.193] 6, 21 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 6, 21 -- X-Sender: hinmeigeng-at-hotmail.com 6, 21 -- Reply-To: R.H.Olley-at-reading.ac.uk 6, 21 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 6, 21 -- To: Microscopy-at-MSA.Microscopy.Com 6, 21 -- Cc: R.H.Olley-at-reading.ac.uk 6, 21 -- Subject: Polyester Fibre Sectioning 6, 21 -- Date: Sat, 01 Apr 2006 10:30:17 +0000 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; format=flowed 6, 21 -- X-OriginalArrivalTime: 01 Apr 2006 10:30:20.0902 (UTC) FILETIME=[471A6860:01C65577] ==============================End of - Headers==============================
==============================Original Headers============================== 20, 31 -- From Colin.Veitch-at-csiro.au Sat Apr 1 15:31:24 2006 20, 31 -- Received: from vic-MTAout5.csiro.au (vic-MTAout5.csiro.au [150.229.64.42]) 20, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k31LVL26020169 20, 31 -- for {Microscopy-at-microscopy.com} ; Sat, 1 Apr 2006 15:31:24 -0600 20, 31 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=Edr5xS+CNVwM1ZkcxZmzJTgVjkzESAVg+fBUD1PNJAo8d4oeaa9LLRw8nWUUBbDf/4eDecwdLsZ/L3OE5ysfljcOqt8MqrZIw1UdUsSO6VdhPf9assrLCiPpf+1hCYjZ; 20, 31 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 20, 31 -- by vic-MTAout5.csiro.au with ESMTP; 02 Apr 2006 07:31:20 +1000 20, 31 -- X-IronPort-AV: i="4.03,154,1141563600"; 20, 31 -- d="scan'208"; a="73618511:sNHT60718480" 20, 31 -- Received: from exvicn2-mel.nexus.csiro.au ([138.194.3.62]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 20, 31 -- Sun, 2 Apr 2006 07:31:19 +1000 20, 31 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exvicn2-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 20, 31 -- Sun, 2 Apr 2006 07:31:19 +1000 20, 31 -- X-MIMEOLE: Produced By Microsoft Exchange V6.0.6603.0 20, 31 -- Content-Class: urn:content-classes:message 20, 31 -- MIME-Version: 1.0 20, 31 -- Content-Type: text/plain; 20, 31 -- charset="Windows-1252" 20, 31 -- Subject: RE: [Microscopy] Polyester Fibre Sectioning 20, 31 -- Date: Sun, 2 Apr 2006 07:31:17 +1000 20, 31 -- Message-ID: {32CDDDAA7161394599F002549491574902480B-at-exvic5-gex.nexus.csiro.au} 20, 31 -- X-MS-Has-Attach: 20, 31 -- X-MS-TNEF-Correlator: 20, 31 -- Thread-Topic: [Microscopy] Polyester Fibre Sectioning 20, 31 -- Thread-Index: AcZVd07U+l155sLxTiK9SMcIEvwpgAAW0W4b 20, 31 -- References: {200604011030.k31AUT1P011648-at-ns.microscopy.com} 20, 31 -- From: {Colin.Veitch-at-csiro.au} 20, 31 -- To: {hinmeigeng-at-hotmail.com} , {Microscopy-at-microscopy.com} 20, 31 -- X-OriginalArrivalTime: 01 Apr 2006 21:31:19.0827 (UTC) FILETIME=[9DAA1E30:01C655D3] 20, 31 -- Content-Transfer-Encoding: 8bit 20, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k31LVL26020169 ==============================End of - Headers==============================
For general reference and because I'm just curious, what is an "injector blade" and when would you use it in a microtome instead of either diamond or glass blades.
Or is this blade not used with microtomes?
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 6, 11 -- From zaluzec-at-microscopy.com Sun Apr 2 12:22:12 2006 6, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k32HMBkP025186 6, 11 -- for {microscopy-at-microscopy.com} ; Sun, 2 Apr 2006 12:22:11 -0500 6, 11 -- Mime-Version: 1.0 6, 11 -- Message-Id: {p06110401c055b822c831-at-[206.69.208.22]} 6, 11 -- Date: Sun, 2 Apr 2006 12:22:10 -0500 6, 11 -- To: microscopy-at-microscopy.com 6, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 6, 11 -- Subject: Injector Blade? 6, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Some of you listers enjoy using the inexpensive children's digital microscope, the QX3, as a home hobby item. It's gone thru several changes, but it's still available, with improved software. There's a new software package, available at http://www.edhsw.com/mixscope/index.html
I have no interest in the company, and no personal opinion on the product; I just want you to have fun... -- 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
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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
One can use disposable injector blades from a dispenser quite similar to what used to be used for razor blades for shaving in old-fashioned packages for either paraffin microtomy or cryostat work. I do not believe there is an application for ultramicrotomy where they would take the place of a diamond or glass blade. Tom
At 12:23 PM 04/02/06, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Ah yes, how fondly i remember my old Schick injector. First razor - took it to university with me. Then they got chromium edged blades, Then someone decided we needed two blades, and the rest was history. Now we can get 5 blades, with a motor of some sort, too. In spite of the nostalgia, must confess that it did cut my face up all to heck and gone. But i still have the old razor....
While I'm tired of cutting my face up, I still use injector blades in the blade holder for my old ultratome III to trim blocks.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Phone: 204-789-3313 Pager: 204-931-954 Home Phone: 204-489-6924 Cell: 204-781-1502 Fax: 204-789-3926/204-489-6924
Take a look at the Tips page on our web site (set out below) to see a technique that we have used for many years to determine the TRUE cross section of a fibre or similar material.
I do not believe that any cutting method, other than embedding and sectioning, truly demonstrates the cross section of a material, fracturing does!
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7802 966067 Web www.emcourses.com
----- Original Message ----- X-from: {hinmeigeng-at-hotmail.com} To: {protrain-at-emcourses.com} Sent: Saturday, April 01, 2006 11:31 AM
Nestor,
An injector blade is something we bearded types have long happily forgotten about. It's the narrow, single-edge razor blade that used to go into injector-razors. The main competitor to double-edge razors Way Back When. The "injector" bit was because of how they were supplied: in a little metal carrier with a key that slid into the back of the razor. The blade was then pushed into the razor ("injected"), simultaneously pushing out the old blade. I used Schick Platinum-Plus injector blades in a Vibratome, they worked much better than the commerical microscopy-supply house microtome blades and were much cheaper.
Phil
} For general reference and because I'm just curious, what is an } "injector blade" and when would you use it in a microtome } instead of either diamond or glass blades. } } Or is this blade not used with microtomes? } } } } Nestor } Your Friendly Neighborhood SysOp -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Mon Apr 3 07:36:34 2006 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k33CaYxB010877 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 07:36:34 -0500 4, 22 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k33DE14p031280 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 09:14:07 -0400 4, 22 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 22 -- Mon, 3 Apr 2006 08:36:08 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230903c056c6481aa4-at-[141.209.160.132]} 4, 22 -- In-Reply-To: {200604021728.k32HSSGX000601-at-ns.microscopy.com} 4, 22 -- References: {200604021728.k32HSSGX000601-at-ns.microscopy.com} 4, 22 -- Date: Mon, 3 Apr 2006 08:36:28 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] Injector Blade? 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 03 Apr 2006 12:36:08.0741 (UTC) FILETIME=[2ECABD50:01C6571B] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -4 () L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Thanks to all who responded to my inquiry regarding the Replacement valve. I was able to find several sources for adequate replacements. The key was adding the term "metering valve" to my growing vocabulary. With the proper name and an internet connection, it only took a few minutes to find exactly what I needed.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 3, 23 -- From hagglundk1-at-nku.edu Mon Apr 3 08:20:04 2006 3, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k33DK4Qs021072 3, 23 -- for {microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 08:20:04 -0500 3, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 3, 23 -- Mon, 3 Apr 2006 09:20:02 -0400 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 3, 23 -- Content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="us-ascii" 3, 23 -- Subject: Re: [Microscopy] Replacement Argon Valve THANKS 3, 23 -- Date: Mon, 3 Apr 2006 09:20:01 -0400 3, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3586D1-at-mailfac1.hh.nku.edu} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: Re: [Microscopy] Replacement Argon Valve THANKS 3, 23 -- Thread-Index: AcZXIVBc5msXIo1MShuTFFZwXIUP9A== 3, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 3, 23 -- To: {microscopy-at-microscopy.com} 3, 23 -- X-OriginalArrivalTime: 03 Apr 2006 13:20:02.0668 (UTC) FILETIME=[50BC1AC0:01C65721] 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k33DK4Qs021072 ==============================End of - Headers==============================
For those of you who haven't been able to visualize this blade, Here is a link to an old ad by Schick. I haven't been able to find a link to the original AO blade holder that used these blades, but I do have two in my lab that we are still using, both for paraffin and frozen sections. We do get strange looks at the local pharmacy, though, when we ask for the blades.
http://www.rareads.com/scans/8792.jpg
Joel
} } } } ---------------------------------------------------------------------- } ------ 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 } ---------------------------------------------------------------------- } ------ } } } For general reference and because I'm just curious, what is an } "injector blade" and when would you use it in a microtome } instead of either diamond or glass blades. } } Or is this blade not used with microtomes? } } } } Nestor } Your Friendly Neighborhood SysOp } } ==============================Original } Headers============================== 6, 11 -- From } zaluzec-at-microscopy.com Sun Apr 2 12:22:12 2006 6, 11 -- Received: } from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 11 -- by } ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k32HMBkP025186 6, 11 -- for {microscopy-at-microscopy.com} ; Sun, 2 Apr } 2006 12:22:11 -0500 6, 11 -- Mime-Version: 1.0 6, 11 -- Message-Id: } {p06110401c055b822c831-at-[206.69.208.22]} 6, 11 -- Date: Sun, 2 Apr 2006 } 12:22:10 -0500 6, 11 -- To: microscopy-at-microscopy.com 6, 11 -- From: } "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 6, 11 -- Subject: } Injector Blade? 6, 11 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
In my younger days, when the 2 blade razors were being advertised, I was watching "Saturday Night Live". They would do some spoof ads, and they would do a pretty good job of it. You could hardly tell the difference between their ads, and the real ones. They put together a great ad for a 3 bladed razor that looked pretty real, compared to the 2 blade razor ads. At the end of the ad, they ended with the line, "Because you'll believe anything!" Years later, I saw a real ad for the triple edged razor, and that old "Saturday Night Live" ad came back to me. Now they are selling 5 edged razors! They must work well, because they are selling. I wish I had patented the idea all those years ago...
Have a good day (or evening)!
Darrell
paul_hazelton-at-umanitoba.ca wrote on 04/02/2006 09:26:13 PM:
} 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 } ----------------------------------------------------------------------------
} } Ah yes, how fondly i remember my old Schick injector. First razor - } took it to university with me. Then they got chromium edged blades, } Then someone decided we needed two blades, and the rest was history. } Now we can get 5 blades, with a motor of some sort, too. In spite of } the nostalgia, must confess that it did cut my face up all to heck and } gone. But i still have the old razor.... } } } While I'm tired of cutting my face up, I still use injector blades in } the blade holder for my old ultratome III to trim blocks. } } } paul } } } } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } e-mail: paul_hazelton-at-umanitoba.ca } Work Phone: 204-789-3313 } Pager: 204-931-954 } Home Phone: 204-489-6924 } Cell: 204-781-1502 } Fax: 204-789-3926/204-489-6924 } } } ==============================Original Headers============================== } 10, 21 -- From paul_hazelton-at-umanitoba.ca Sun Apr 2 20:24:51 2006 } 10, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc. } umanitoba.ca [130.179.16.23]) } 10, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k331OovE014494; } 10, 21 -- Sun, 2 Apr 2006 20:24:50 -0500 } 10, 21 -- Received: from [130.179.152.71] (cvx-008.cc.umanitoba.ca } [130.179.152.71]) } 10, 21 -- (authenticated bits=0) } 10, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP } id k331OkFm012539; } 10, 21 -- Sun, 2 Apr 2006 20:24:48 -0500 (CDT) } 10, 21 -- Message-ID: {4430795C.9060407-at-umanitoba.ca} } 10, 21 -- Date: Sun, 02 Apr 2006 20:24:44 -0500 } 10, 21 -- From: Paul Hazelton {paul_hazelton-at-umanitoba.ca} } 10, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en- } US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) } 10, 21 -- X-Accept-Language: en-us, en } 10, 21 -- MIME-Version: 1.0 } 10, 21 -- To: zaluzec-at-microscopy.com } 10, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} } 10, 21 -- Subject: Re: [Microscopy] Injector Blade? } 10, 21 -- References: {200604021725.k32HP0bj028969-at-ns.microscopy.com} } 10, 21 -- In-Reply-To: {200604021725.k32HP0bj028969-at-ns.microscopy.com} } 10, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 10, 21 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 27 -- From milesd-at-us.ibm.com Mon Apr 3 11:07:58 2006 9, 27 -- Received: from e3.ny.us.ibm.com (e3.ny.us.ibm.com [32.97.182.143]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k33G7wbU010192 9, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 3 Apr 2006 11:07:58 -0500 9, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 9, 27 -- by e3.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k33G7lZx008773 9, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 3 Apr 2006 12:07:47 -0400 9, 27 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 9, 27 -- by d01relay02.pok.ibm.com (8.12.10/NCO/VER6.8) with ESMTP id k33G7arN249918 9, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 3 Apr 2006 12:07:37 -0400 9, 27 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 9, 27 -- by d01av04.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k33G7arZ012435 9, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 3 Apr 2006 12:07:36 -0400 9, 27 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 9, 27 -- by d01av04.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k33G7a3T012414 9, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 3 Apr 2006 12:07:36 -0400 9, 27 -- In-Reply-To: {200604030126.k331QDaD015919-at-ns.microscopy.com} 9, 27 -- Subject: Re: [Microscopy] Re: Injector Blade? 9, 27 -- To: Microscopy-at-MSA.Microscopy.Com 9, 27 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 9, 27 -- Message-ID: {OFF56D0893.E91E22F3-ON85257145.0054402D-85257145.0058956C-at-us.ibm.com} 9, 27 -- From: Darrell Miles {milesd-at-us.ibm.com} 9, 27 -- Date: Mon, 3 Apr 2006 12:07:34 -0400 9, 27 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 6.5.4FP3 HF3|February 22, 2006) at 9, 27 -- 04/03/2006 12:07:35 9, 27 -- MIME-Version: 1.0 9, 27 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Hello, I meant to look this up over the weekend, but my internet and phone was down. I thought I would ask this here first.
I wanted to find papers or references on the infinite focus types of stereo microscopes that combine in-focus regions of a series of images into one image. A prodoct that is commercially available that is similar is from alicona imaging called an infinite focus microscope.
Can anyone forward me references or journal references? Any guidance greatly appreciated. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 4, 24 -- From gvrdolja-at-nature.berkeley.edu Mon Apr 3 12:12:12 2006 4, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k33HCBSh020695 4, 24 -- for {microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 12:12:12 -0500 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 18394C1E70 4, 24 -- for {microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 10:12:11 -0700 (PDT) 4, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 4, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 4, 24 -- with ESMTP id 15229-04-2 for {microscopy-at-microscopy.com} ; 4, 24 -- Mon, 3 Apr 2006 10:12:10 -0700 (PDT) 4, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 4, 24 -- id A9935C1E5C; Mon, 3 Apr 2006 10:12:10 -0700 (PDT) 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 94CB7C1E57 4, 24 -- for {microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 10:12:10 -0700 (PDT) 4, 24 -- Date: Mon, 3 Apr 2006 10:12:10 -0700 (PDT) 4, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 4, 24 -- To: microscopy-at-microscopy.com 4, 24 -- Subject: question about infinite focus stereo microscopes 4, 24 -- Message-ID: {Pine.SOC.4.64.0604031008050.2468-at-nature.Berkeley.EDU} 4, 24 -- MIME-Version: 1.0 4, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 24 -- X-Virus-Scanned: amavisd-new at nature.berkeley.edu ==============================End of - Headers==============================
The Final Program for SCANNING 2006 is now available at www.scanning.org. The Registration Form is also available online and we encourage you to register today to ensure your first choice for Short Courses and Sessions.
We look forward to seeing you April 25-27 at the Hotel Washington in Washington, D.C.
The blades are still available through microscopy supply houses. I use them for many purposes myself. Maybe someone already pointed this out, but I thought I'd mention it since it sounds like some people are still looking for a source. There is even a picture of the dispenser on the Ted Pella site/catalog.
On 3 Apr 2006 at 10:12, jbs-at-temple.edu wrote:
} For those of you who haven't been able to visualize this blade, Here } is a link to an old ad by Schick. I haven't been able to find a link } to the original AO blade holder that used these blades, but I do have } two in my lab that we are still using, both for paraffin and frozen } sections. We do get strange looks at the local pharmacy, though, } when we ask for the blades. } } } http://www.rareads.com/scans/8792.jpg } } Joel
All the best,
Andy Buechele, Washington, D.C., U.S.A.
==============================Original Headers============================== 6, 22 -- From andrewb-at-mail.vsl.cua.edu Mon Apr 3 13:01:36 2006 6, 22 -- Received: from hermes.vsl.cua.edu (interface.vsl.cua.edu [136.242.188.2]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k33I1XEC007894 6, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 13:01:34 -0500 6, 22 -- X-Filtered-With-Copfilter: Version 0.82 (ProxSMTP 1.3.91) 6, 22 -- X-Copfilter-Virus-Scanned: ClamAV 0.88/1370 - Mon Apr 3 12:31:59 2006 6, 22 -- X-Copfilter: Client is part of our network, skipped SpamAssassin 6, 22 -- Received: from andrewb_409 [136.242.189.249] by hermes.vsl.cua.edu with ESMTP 6, 22 -- (SMTPD-8.21) id A2FC0350; Mon, 03 Apr 2006 14:01:32 -0400 6, 22 -- From: "Andrew Buechele" {andrewb-at-vsl.cua.edu} 6, 22 -- To: Microscopy-at-microscopy.com 6, 22 -- Date: Mon, 03 Apr 2006 14:01:32 -0400 6, 22 -- MIME-Version: 1.0 6, 22 -- Subject: Re: [Microscopy] Re: Injector Blade? 6, 22 -- Reply-to: andrewb-at-vsl.cua.edu 6, 22 -- Message-ID: {44312ABC.32465.75EBF0-at-localhost} 6, 22 -- Priority: normal 6, 22 -- In-reply-to: {200604031512.k33FCsmB008434-at-ns.microscopy.com} 6, 22 -- X-mailer: Pegasus Mail for Windows (v4.02a) 6, 22 -- Content-type: text/plain; charset=US-ASCII 6, 22 -- Content-transfer-encoding: 7BIT 6, 22 -- Content-description: Mail message body ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both chao.wang-at-materials.ox.ac.uk as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: chao.wang-at-materials.ox.ac.uk Name: Chao Wang
Organization: Oxford Materials
Title-Subject: [Filtered] help for MgO prepration and Fe oxidation
Question: Dear All
I have three problems, could you tell me some suggestions:
1.single crystale MgO preparation
I need very good quality cross section TEM sample. I now manully polish it to 70 micron and dimple to abount 20 micron and then ion milling (PIPS). But it's not so good. The edge looks sharp and MgO cracks a lot when I ground lower then 60 micron on diamond papers. Do you have better ideas to get very thin specimen?
2. Fe oxidiation Another problem is Fe oxidation (on top of MgO substrate), it seems after some time, the sample oxidized a lot, even I got very thin specimen, it looks like amorphous (it should be crystalline). How to keep it? (I use plasma cleaning, not working very well)
3. Beam sensitive to MgO sample
Sometime the beam in TEM is sensitive to MgO sample, how to get rid of this?
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rra-at-stowers-institute.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
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
I too have experienced similar problems in preparing thin specimens from single crystal MgO. I have had some success with plan view specimens ie ion milling away the substrate to perforate through into a thin oxide film grown on it (See J. Cryst. Growth 285 (2005) p208-214). Here samples were ground to about 80µms thick, then dimpled. I never get too adventurous with dimpling - aiming for a thickness of 30µms. Any thinner and the MgO has a strong inclination to fracture. All grinding and dimpling is done very gently to avoid cracking the substrate. I then ion mill in a PIPs at 5keV, 6 deg, until the sample is near perforation, then reduce the angle to 4 deg and the voltage to 3keV and then mill to electron transparency. To liberate the specimen from the mounting post I soak it in acetone till it drops off rather than heating to soften the crystal bond and slide it off - the latter is guaranteed to break thin regions off. Tripod Polishing was of no use at all, aside from the coarse grinding step to get down to about 80µms.
Success was very hit and miss. I suspect my MgO crystals had a high degrees of residual stress. Cross sectioning is even harder and requires a lot of patience to get any kind of result. Specimens just disintegrate in the PIPS without any mechanical handling. Focused ion beam milling may help if you have access to one.
With regard to oxidation of Fe. My experience is with electropolished foils rather than thin films, but there are some similarities. Storage in solvents like methanol is definitely not recommended - such polar solvents are highly corrosive to iron. Storage in a non-polar solvent like a hydrocarbon may stop oxidation, but cleaning it up for TEM examination could be challenge without a plasma cleaner. Storage in vacuum is surprisingly bad for electropolished foils and they oxidise much more severely than storage in a normal lab desiccator. I suspect the protective hydrated film formed from electropolishing dries out in vacuum and cracks allowing oxidation to proceed. Of course your films don't have such a layer, so vacuum storage may be no worse or better than a good lab desiccator. Probably the key factor governing oxidation is the level of water vapour your specimen gets exposed to.
With respect to beam sensitivity - I presume you mean charging rather than beam damage? At 200keV I did not experience significant damage in the MgO substrate, but electron beam charging was a major issue. I therefore always evaporate a very thin layer of carbon (20Å) onto MgO specimens prior to TEM examination. If the film grown on the MgO is not very conductive, then I coat both sides. In your case you have a conductive layer on one side (Fe), so you may get away with coating just the MgO side.
I hope this helps.
Regards
Dave Mitchell -- Dr David Mitchell ANSTO Materials PMB 1 Menai NSW 2234 Australia
tel 61 2 9717 3456 fax 61 2 9543 7179
==============================Original Headers============================== 11, 25 -- From drm-at-ansto.gov.au Mon Apr 3 23:52:41 2006 11, 25 -- Received: from tachyon.gw.ansto.gov.au (anstogw.gw.ansto.gov.au [137.157.8.253]) 11, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k344qejx015539 11, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 23:52:41 -0500 11, 25 -- Received: (from uucp-at-localhost) 11, 25 -- by tachyon.gw.ansto.gov.au (8.11.7p1+Sun/8.11.7) id k344qcC15261 11, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 14:52:38 +1000 (EST) 11, 25 -- Received: from jenner.ansto.gov.au(137.157.59.25) by tachyon.gw.ansto.gov.au via csmap (V6.0) 11, 25 -- id srcAAARlaGZD; Tue, 4 Apr 06 14:52:38 +1000 11, 25 -- Received: from hadron.ansto.gov.au (hadron.ansto.gov.au [137.157.13.219]) 11, 25 -- by jenner.ansto.gov.au (8.12.10/8.12.10) with ESMTP id k344qXk2006413 11, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 14:52:33 +1000 11, 25 -- Received: from [137.157.94.26] (m0374m.ansto.gov.au [137.157.94.26]) 11, 25 -- by hadron.ansto.gov.au (8.9.3/8.9.3) with ESMTP id OAA06227 11, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 14:52:31 +1000 (EST) 11, 25 -- Mime-Version: 1.0 11, 25 -- Message-Id: {a06230914c057a30b5d12-at-[137.157.94.26]} 11, 25 -- Date: Tue, 4 Apr 2006 14:52:37 +1000 11, 25 -- To: Microscopy-at-microscopy.com 11, 25 -- From: David Mitchell {drm-at-ansto.gov.au} 11, 25 -- Subject: help for MgO prepration and Fe oxidati 11, 25 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 11, 25 -- X-ANSTO-MailScanner: Found to be clean 11, 25 -- Content-Transfer-Encoding: 8bit 11, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k344qejx015539 ==============================End of - Headers==============================
I have a Philips 410 TEM which has decided not to cooperate! I am not overly familiar with this machine so I apologise if I have missed something obvious.
When you turn on the system and begin the pumpdown sequence the rotary pump starts and after around 30 seconds (the vacuum gauge drops to around 10 on the scale) the rotary pump shuts off and that is it.
The vacuum system indicators (valve status) on the side panel all remain off. I have checked all the fuses and they are OK. There is plenty of cooling water (going in and coming out). The diffusion pump heater seems to be OK (it is not open circuit, nor ohms resistance). The pneumatic gas pressure is also OK.
Any help as to a possible cause/cure would be appreciated.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
==============================Original Headers============================== 17, 30 -- From Colin.Veitch-at-csiro.au Tue Apr 4 00:20:22 2006 17, 30 -- Received: from vic-MTAout2.csiro.au (vic-MTAout2.csiro.au [150.229.64.38]) 17, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k345KLVZ025429 17, 30 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 00:20:21 -0500 17, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=dBF7dfKXSlt81lkUABrPqUbsCIwjoURW1HJ9hzb0jZaxVI1s/fKCKoicAEX1Lot9kXcu1OaMU4mhpGKGJD9rj0Yt5OO80G/XIVKO1w6MjYmSjaEgTZ03epxNTfKPHGmp; 17, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 17, 30 -- by vic-MTAout2.csiro.au with ESMTP; 04 Apr 2006 15:20:20 +1000 17, 30 -- X-IronPort-AV: i="4.03,160,1141563600"; 17, 30 -- d="scan'208"; a="73879638:sNHT25511800" 17, 30 -- Received: from exvicn1-mel.nexus.csiro.au ([138.194.3.60]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 17, 30 -- Tue, 4 Apr 2006 15:20:19 +1000 17, 30 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exvicn1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 17, 30 -- Tue, 4 Apr 2006 15:20:19 +1000 17, 30 -- X-MIMEOLE: Produced By Microsoft Exchange V6.0.6603.0 17, 30 -- Content-Class: urn:content-classes:message 17, 30 -- MIME-Version: 1.0 17, 30 -- Content-Type: text/plain; 17, 30 -- charset="us-ascii" 17, 30 -- Subject: Problem with a Philips EM410 TEM 17, 30 -- Date: Tue, 4 Apr 2006 15:20:18 +1000 17, 30 -- Message-ID: {32CDDDAA7161394599F0025494915749024813-at-exvic5-gex.nexus.csiro.au} 17, 30 -- X-MS-Has-Attach: 17, 30 -- X-MS-TNEF-Correlator: 17, 30 -- Thread-Topic: Problem with a Philips EM410 TEM 17, 30 -- Thread-Index: AcZXp3YO7P7oXMhtRkmVma7B0e0d9A== 17, 30 -- From: {Colin.Veitch-at-csiro.au} 17, 30 -- To: {microscopy-at-microscopy.com} 17, 30 -- X-OriginalArrivalTime: 04 Apr 2006 05:20:19.0708 (UTC) FILETIME=[772AB3C0:01C657A7] 17, 30 -- Content-Transfer-Encoding: 8bit 17, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k345KLVZ025429 ==============================End of - Headers==============================
please contact me offline. I can send you information on how you can combine a stack of stereomicroscope images and get a result image with "infinite" focus.
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
We have a Leica EM-Stain. We purchased it about a year ago in a quest to make our staining absolutely consistent, and in regard to that, it works wonderfully. Given all of the options for wetting times, stain times, stain temperatures, etc, it's pretty versatile and dead-on accurate, not to mention it frees up a lot of a technician's time! However, we're had nothing but problems with the unit. We've gone though 3 pumps already, numerous service calls, etc. In Leica's defense, they may have had a bad batch of stain (high amounts of precipitate), which is clogging up the pump and the filters. Speaking of the stain, you're "required" to use their pre-made stains, else you void the warranty on the unit. There's a bit of upkeep with the stainer as well, in that you must perform a cleaning cycle at least once a week (which only takes a few minutes to get running but someone still has to remember to do it), and it uses 3% Nitric acid for the cleaning cycle, which possibly adds another waste stream for your lab. All in all, it would be a great unit if there weren't so many problems with it. We may just have ended up with a lemon, though. Hard to say.
-Chris
rra-at-stowers-institute.org 04/03/2006 10:20 PM Please respond to rra
To: Christopher Hayden/PH/Novartis-at-PH cc: Subject: [Microscopy] viaWWW: Leica EM-Stain
---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rra-at-stowers-institute.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
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
I don't know the Philips 410, so this may be useless, but I did have a similar problem with a JEOL SEM, and the cure I think may be generic. The issue is that if a given vacuum, as registered on the vacuum control board as a given current (e.g. {50mA) or voltage (e.g.??), is not shown, the system doesn't switch to the diff pump, but shuts down. The cure was to go to manual pumping and allow the 'scope to pump with the rotary pump for longer than the electronics would allow (usually 2 to 5 minutes). Then manually go to high vacuum and let run for a day or two. It should then cycle OK. This was after the 'scope had been sitting with a static vacuum for a few weeks. There should be a test point on the vacuum board where the required current (voltage) can be read, and this should be in the manual somewhere. Servicing the vacuum system or the like, maybe troubleshooting. The other thought is, it's after April Fool's day, do you have any pranksters in the department? In a previous incarnation, I had a service engineer for a now-defunct EM company try to sell me a new diff pump (for $12,000) by placing a nickel between the heater and the pump bottom. The pump didn't heat as well, obviously, so the vacuum was degraded, and there was a cheery red glow from the bottom of the pump. (No, they didn't sell a pump.)
Phil Now I know Oz is different -- you give your mobs numbers, and we give our gangs names.
} Hi, } } I have a Philips 410 TEM which has decided not to cooperate! I am not } overly familiar with this machine so I apologise if I have missed } something obvious. } } When you turn on the system and begin the pumpdown sequence the rotary } pump starts and after around 30 seconds (the vacuum gauge drops to } around 10 on the scale) the rotary pump shuts off and that is it. } } The vacuum system indicators (valve status) on the side panel all remain } off. I have checked all the fuses and they are OK. There is plenty of } cooling water (going in and coming out). The diffusion pump heater } seems to be OK (it is not open circuit, nor ohms resistance). The } pneumatic gas pressure is also OK. } } Any help as to a possible cause/cure would be appreciated. } } Cheers. } } Colin Veitch } Electron Microscopist } CSIRO Textile and Fibre Technology } PO Box 21, BELMONT, Vic. 3216. Australia. } E-mail: colin.veitch-at-csiro.au } Web: http://www.tft.csiro.au } Tel: +61 (0) 3 5246 4000 } Mob: 0438 538 475 } Fax: +61 (0) 3 5246 4811 } } The information contained in this e-mail message may be privileged or } confidential information. If you are not an intended recipient, you may } not copy, distribute or take any action in reliance on it. If you have } received this message in error, please telephone CSIRO Textile and Fibre } Technology on +61 3 5246 4000. -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 23 -- From oshel1pe-at-cmich.edu Tue Apr 4 07:14:43 2006 4, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k34CEho9027735 4, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 07:14:43 -0500 4, 23 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k34Cpx4p006142; 4, 23 -- Tue, 4 Apr 2006 08:52:08 -0400 4, 23 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 23 -- Tue, 4 Apr 2006 08:14:07 -0400 4, 23 -- Mime-Version: 1.0 4, 23 -- Message-Id: {f06230905c058111a17a7-at-[141.209.160.132]} 4, 23 -- In-Reply-To: {200604040525.k345PRid000860-at-ns.microscopy.com} 4, 23 -- References: {200604040525.k345PRid000860-at-ns.microscopy.com} 4, 23 -- Date: Tue, 4 Apr 2006 08:14:28 -0400 4, 23 -- To: Colin.Veitch-at-csiro.au 4, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 23 -- Subject: Re: [Microscopy] Problem with a Philips EM410 TEM 4, 23 -- Cc: Microscopy-at-microscopy.com 4, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 23 -- X-OriginalArrivalTime: 04 Apr 2006 12:14:07.0069 (UTC) FILETIME=[456D58D0:01C657E1] 4, 23 -- X-CanItPRO-Stream: default 4, 23 -- X-Spam-Score: -4 () L_EXCH_MF 4, 23 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
I don't quite understand the question you are asking about specimen preparation - you say "cross-section", but cross section of what? - so I won't address that part of your post.
I also don't understand where the Fe comes from. However, I do know that over a period of (many) months, MgO samples transform into what appears to be MgCO3. For a period of years, I had a student working on polycystalline MgO. He made samples by ion milling. He did not have to take special precautions with his samples if he examined them days or weeks after they were made, but if we wanted to look at them again a year or two later we had to "tickle" them with the ion mill to clean them up. STEM analysis showed that the carbonate layer had formed. We did not try to prevent this happening as it was not a major problem.
MgO is known to be beam sensitive. A significant part of the damage depends on the current density, so working in a FEG-STEM you can be much worse-off than imaging or using a less intense electron probe. But you can't eliminate the problem. Going to lower bean voltages can (counter-intuitively) make the problem worse, as the cross-section for various electron-sample interactions can actually increase at lower voltages - though the total current in the electron probe will also decrease, which may partially or totally compensate.
Tony Garratt-Reed
} At 10:24 PM 4/3/2006, you wrote:
Email: chao.wang-at-materials.ox.ac.uk Name: Chao Wang
Organization: Oxford Materials
Title-Subject: [Filtered] help for MgO prepration and Fe oxidation
Question: Dear All
I have three problems, could you tell me some suggestions:
1.single crystale MgO preparation
I need very good quality cross section TEM sample. I now manully polish it to 70 micron and dimple to abount 20 micron and then ion milling (PIPS). But it's not so good. The edge looks sharp and MgO cracks a lot when I ground lower then 60 micron on diamond papers. Do you have better ideas to get very thin specimen?
2. Fe oxidiation Another problem is Fe oxidation (on top of MgO substrate), it seems after some time, the sample oxidized a lot, even I got very thin specimen, it looks like amorphous (it should be crystalline). How to keep it? (I use plasma cleaning, not working very well)
3. Beam sensitive to MgO sample
Sometime the beam in TEM is sensitive to MgO sample, how to get rid of this?
Your help is very appreicated!
best wishes
Chao
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both andromeda_tm-at-libero.it as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Cutting angle with a flat razor blade
Question: Hi all,
I wonder if someone could give me some advice about the best cutting angle for specimens (mouse tissues) embedded into wax (Paraplast - melting point 56ƒC) at room temperature (about 18ƒC) with a microtome flat razor blade. Thank you in advance. Best Regards,
Good morning, everybody I am looking for advice on defining resolution and minimal feature size through information theory. In 1992, there was a paper by D. Van Dyck and A.F. de Jong (Ultramicroscopy 47, 266 (1992)), which addressed in some detail general principles on information theory as applied to image formation mechanism in electron microscopy. I am wondering what the current status of this field is. Thank you in advance Sergei
-- Sergei V. Kalinin, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Bldg. 3025, MS6030, Oak Ridge, TN 37831 Phone: (865) 241-0236 FAX: (865) 574-4143 URL: sergei2.kalininweb.com New: http://nanotransport.ornl.gov
==============================Original Headers============================== 2, 30 -- From sergei2-at-ornl.gov Tue Apr 4 08:48:53 2006 2, 30 -- Received: from emroute2.ornl.gov (emroute2.ornl.gov [160.91.86.17]) 2, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k34Dmrqd023746 2, 30 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 08:48:53 -0500 2, 30 -- Received: from emroute2.ornl.gov (localhost [127.0.0.1]) 2, 30 -- by emroute2.ornl.gov (PMDF V6.2-1x9 #31038) 2, 30 -- with ESMTP id {0IX7000HIADGZ1-at-emroute2.ornl.gov} for 2, 30 -- microscopy-at-microscopy.com; Tue, 04 Apr 2006 09:48:53 -0400 (EDT) 2, 30 -- Received: from ORNLEXCHANGE.ornl.gov (ornlexchange2.ornl.gov [160.91.1.22]) 2, 30 -- by emroute2.ornl.gov (PMDF V6.2-1x9 #31038) 2, 30 -- with ESMTP id {0IX700620ADG6P-at-emroute2.ornl.gov} for 2, 30 -- microscopy-at-microscopy.com; Tue, 04 Apr 2006 09:48:52 -0400 (EDT) 2, 30 -- Date: Tue, 04 Apr 2006 09:48:50 -0400 2, 30 -- From: "Kalinin, Sergei V." {sergei2-at-ornl.gov} 2, 30 -- Subject: FW: Resolution and information theory 2, 30 -- To: microscopy-at-microscopy.com 2, 30 -- Message-id: {3E4D6911DB2CA94DA31AC5E1CB522DBB42AEAF-at-ORNLEXCHANGE.ornl.gov} 2, 30 -- MIME-version: 1.0 2, 30 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 2, 30 -- Content-type: text/plain; charset=iso-8859-1 2, 30 -- Importance: high 2, 30 -- Priority: Urgent 2, 30 -- X-Priority: 1 2, 30 -- Thread-Topic: Resolution and information theory 2, 30 -- Thread-Index: AcZX7Nd9csa8AnjnTyq53Z4QkequswAAZdTA 2, 30 -- Content-class: urn:content-classes:message 2, 30 -- X-MS-Has-Attach: 2, 30 -- X-MS-TNEF-Correlator: 2, 30 -- Content-Transfer-Encoding: 8bit 2, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k34Dmrqd023746 ==============================End of - Headers==============================
Based on my experience with a Philips EM 420, I have found the instrument to be very sensitive to having proper water and pneumatic pressure.
I would first check the pneumatics to ensure that it is giving proper pressure. Ours needs about 80 psi, or we get exactly the problem that you describe. Unfortunately, we have that problem a lot because we rely on 'house' air and have no control over the source.
If that doesn't solve the problem, thoroughly examine the water circuit. It may look like "plenty" of water, but it may not be enough. First check and clean the water filter near where the water line enters the back of the scope. It never ceases to surprise me how big a difference that makes!
If that doesn't work, there may be water 'loops' inside that do not have sufficient pressure. The only way to check is to take apart the water lines one at a time and measure the flow of each. (I did that by timing the flow into an open container. The manual will list the specs.) I have found that the isolation values clog over time and need to be replaced. Or do what I did and simply by-pass the faulty valves. If you do that, you will have to manually stop the flow in the correct lines when you do a bake-out.
Good luck!
-- Roger A. Ristau, PhD TEM Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269 vox: 860-486-5379 fax: 860-486-4745
} Hi, } } I have a Philips 410 TEM which has decided not to cooperate! I am not } overly familiar with this machine so I apologise if I have missed } something obvious. } } When you turn on the system and begin the pumpdown sequence the rotary } pump starts and after around 30 seconds (the vacuum gauge drops to } around 10 on the scale) the rotary pump shuts off and that is it. } } The vacuum system indicators (valve status) on the side panel all remain } off. I have checked all the fuses and they are OK. There is plenty of } cooling water (going in and coming out). The diffusion pump heater } seems to be OK (it is not open circuit, nor ohms resistance). The } pneumatic gas pressure is also OK. } } Any help as to a possible cause/cure would be appreciated. } } Cheers. } } Colin Veitch } } Electron Microscopist } } CSIRO Textile and Fibre Technology } } PO Box 21, BELMONT, Vic. 3216. Australia. } } E-mail: colin.veitch-at-csiro.au } } Web: http://www.tft.csiro.au } } Tel: +61 (0) 3 5246 4000 } Mob: 0438 538 475 } Fax: +61 (0) 3 5246 4811 } } } } The information contained in this e-mail message may be privileged or } confidential information. If you are not an intended recipient, you may } not copy, distribute or take any action in reliance on it. If you have } received this message in error, please telephone CSIRO Textile and Fibre } Technology on +61 3 5246 4000. } } } } ==============================Original Headers============================== } 17, 30 -- From Colin.Veitch-at-csiro.au Tue Apr 4 00:20:22 2006 } 17, 30 -- Received: from vic-MTAout2.csiro.au (vic-MTAout2.csiro.au } [150.229.64.38]) } 17, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k345KLVZ025429 } 17, 30 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 00:20:21 -0500 } 17, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; } b=dBF7dfKXSlt81lkUABrPqUbsCIwjoURW1HJ9hzb0jZaxVI1s/fKCKoicAEX1Lot9kXcu1OaMU4mh } pGKGJD9rj0Yt5OO80G/XIVKO1w6MjYmSjaEgTZ03epxNTfKPHGmp; } 17, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) } 17, 30 -- by vic-MTAout2.csiro.au with ESMTP; 04 Apr 2006 15:20:20 +1000 } 17, 30 -- X-IronPort-AV: i="4.03,160,1141563600"; } 17, 30 -- d="scan'208"; a="73879638:sNHT25511800" } 17, 30 -- Received: from exvicn1-mel.nexus.csiro.au ([138.194.3.60]) by } exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); } 17, 30 -- Tue, 4 Apr 2006 15:20:19 +1000 } 17, 30 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by } exvicn1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); } 17, 30 -- Tue, 4 Apr 2006 15:20:19 +1000 } 17, 30 -- X-MIMEOLE: Produced By Microsoft Exchange V6.0.6603.0 } 17, 30 -- Content-Class: urn:content-classes:message } 17, 30 -- MIME-Version: 1.0 } 17, 30 -- Content-Type: text/plain; } 17, 30 -- charset="us-ascii" } 17, 30 -- Subject: Problem with a Philips EM410 TEM } 17, 30 -- Date: Tue, 4 Apr 2006 15:20:18 +1000 } 17, 30 -- Message-ID: } {32CDDDAA7161394599F0025494915749024813-at-exvic5-gex.nexus.csiro.au} } 17, 30 -- X-MS-Has-Attach: } 17, 30 -- X-MS-TNEF-Correlator: } 17, 30 -- Thread-Topic: Problem with a Philips EM410 TEM } 17, 30 -- Thread-Index: AcZXp3YO7P7oXMhtRkmVma7B0e0d9A== } 17, 30 -- From: {Colin.Veitch-at-csiro.au} } 17, 30 -- To: {microscopy-at-microscopy.com} } 17, 30 -- X-OriginalArrivalTime: 04 Apr 2006 05:20:19.0708 (UTC) } FILETIME=[772AB3C0:01C657A7] } 17, 30 -- Content-Transfer-Encoding: 8bit } 17, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k345KLVZ025429 } ==============================End of - Headers==============================
==============================Original Headers============================== 12, 20 -- From raristau-at-ims.uconn.edu Tue Apr 4 08:55:32 2006 12, 20 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k34DtWZC031023 12, 20 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 08:55:32 -0500 12, 20 -- Received: from [137.99.44.238] (d44h238.public.uconn.edu [137.99.44.238]) 12, 20 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k34DtHDJ031700 12, 20 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 09:55:18 -0400 12, 20 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 12, 20 -- Date: Tue, 04 Apr 2006 09:54:52 -0400 12, 20 -- Subject: Re: [Microscopy] Problem with a Philips EM410 TEM 12, 20 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 12, 20 -- To: {microscopy-at-microscopy.com} 12, 20 -- Message-ID: {C057F2EC.B25%raristau-at-ims.uconn.edu} 12, 20 -- In-Reply-To: {200604040525.k345PveJ002113-at-ns.microscopy.com} 12, 20 -- Mime-version: 1.0 12, 20 -- Content-type: text/plain; charset="US-ASCII" 12, 20 -- Content-transfer-encoding: 7bit 12, 20 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 12, 20 -- X-UConn-MailScanner: Found to be clean 12, 20 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
Caroline has usefully pointed out that 'miXscope' software is now available for the fun 'Digital Blue' QX5 children's microscope, which is an upgrade of the older 'Intel' QX3 plastic microscope. Both are used in many homes and schools. I suspect Caroline is an Apple user, as the highly regarded miXscope software allows you to use the QX-5 (and the QX-3 and the MiScope USB) with an Apple G3 to G5 computer - for $15 single, $20 schoolwide.
The QX-5 [and QX-3] is supplied with Windows only software and otherwise won't work at all with an Apple MAC. At present miXscope doesn't seem to support the new Intel version Apples though, and it requires Mac OS X 10.2.8 or later.
For those interested, I attach my 'Amazon.co.uk' review of the kid friendly QX-5 microscope, based on my experience with my kid's one. The QX-5 site is at http://www.playdigitalblue.com. Plus there's the superb micro.magnet.fsu.edu/optics/intelplay site for the QX-3 (they are promising to update it for the QX-5). There is also some QX-5 help forum at http://www.expansys.com.
Regards
Keith
My amazon.co.uk Review of the Digital Blue QX-5 'microscope'.
This is a rather fun toy microscope that has a built in CMOS detector so that images can only be viewed via a Windows PC. The all plastic construction (including lenses) limits the accuracy of focussing and the on-screen image resolution is adequate rather than good. This microscope was originally marketed by Intel and built by toy manufacturer Mattel as the QX-3. Now Digital Blue have taken it on after Intel discontinued production. The QX-5 is an upgrade having 640 x 480 pixel resolution rather than just 352 x 288 in the original QX-3. Have a look at micro.magnet.fsu.edu/optics/intelplay for very detailed scientific description of the original QX-3 and advice on what to use it for. Every school in the UK was given one of these in 2002.
I installed the QX-5 software under Windows XP Pro on a 1.2MHz Athlon PC and the software worked fine. The only downside is that the software changes the CRT screen refresh rate to 60Hz and doesn't switch it back to the flicker free 85Hz. So a trip to 'Start, Control Panel, Display, Settings, Advanced, Monitor' is required to set the graphics back to their correct setting (check these before you run the software). Otherwise the software and USB microscope run very well. It comes with a small prepared 'slide' (a cardboard and plastic array of things like insect parts) plus a reasonable archive of digital images which you can add to.
Once on the PC the 640x480 images can be manipulated and pasted etc, and it does time-lapse for things like crystal growth (but there's not much control of the time-lapse intervals). I have a QX-5 at home for the kids, but like most kids with microscopes they can get bored with it after running out of things to view - so web and book searches for ideas is useful.
The QX-5 has not got the resolution of even a standard 'school' compound microscope though, largely because you see it all 'enlarged' on a large computer screen, it uses plastic lenses and has a low resolution detector (but you can share the view with friends). So you may find the QX-5 a real disappointment if you expect too much of it in terms of image quality. However it is rather fun to use and has transmission + reflection white LED light sources built in to view specimens. The software is also very kid friendly and the increased resolution over the QX-3 is very welcome. So overall, recommended for pre-teen budding scientists.
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: schooley-at-mcn.org [mailto:schooley-at-mcn.org] Sent: 02 April 2006 19:04 To: keith.morris-at-ucl.ac.uk
Some of you listers enjoy using the inexpensive children's digital microscope, the QX3, as a home hobby item. It's gone thru several changes, but it's still available, with improved software. There's a new software package, available at http://www.edhsw.com/mixscope/index.html
I have no interest in the company, and no personal opinion on the product; I just want you to have fun... -- 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
I have a student who is looking at particles with a polystyrene core and a polyamine shell. He is interested in staining the particles so that he can distinguish the two layers. Presently the particles are on the order of 0.5 micron diameter but he is hoping to reduce this down to a few tens of nm.
It sounds like he has some references that used either OsO4 or RuO4 as stains.
I understand that these stains are commonly used in the biology world, I am a materials person and have no experience with these. I'm looking for a couple pieces of info:
1) Recommendations for preferentially staining either the core or the shell. (With the above stains or something else)
2) Cautions for using these stains, I understand they are quite toxic.
3) A reference which discusses all this would be great.
The student talks about looking at these in the SEM, but I think the TEM will be more effective, specially if he manages to get his particles as small as he wants. Is there a way to image the core vs the shell in the SEM? I have a FESEM so it might be possible to image the small particles he anticipates, but I don't think I can get an image of the core through a 0.125 micron shell. Thoughts?
Thanks for your help.
Tom
------------------------------------------------------------------------ --------------------------------------------- Thomas M Murray email: murraytm-at-u.washington.edu Electron Microscopy Center Manager Phone: (206)543-2836 Materials Science & Engineering Fax: (206)543-3100 Box 352120 302 Roberts Hall Cell: (425)345-0083 University of Washington Seattle, WA 98195
I think that I can address all of your questions to some extent:
1) For the preparation of MgO by ion milling, you might consider the sequence of sample preparation espoused by Arpa Barna and Technoorg-Linde where samples are ground very thin in special titanium grids made for cross section and using low angle milling. The complete process is given in The Handbook of Microscopy, Applications in Materials Science, Solid-State Physics, and Chemistry, edited by S. Amelinckx, D. van Dyck, J. van Landuyt, G.van Tendeloo, and published by VCH. Technoorg-Linde has a suite of instruments that are very useful for cutting and mounting the samples for this process. This technique will minimize surface thickness variations of different materials in a cross section due to the geometrical factors affecting ion milled samples.
I know that I have tried the small angle cleavage technique with MgO in the past, but I can't remember whether I had success with it or not. If anyone has an example of a TEM sample prepared by cleaving single crystal MgO, I would be particularly interested in see those results. If you are interested, we have the MicroCleave(TM) sample preparation kit available.
2) It is not surprising that a vacuum does not stop the oxidation process of Fe. Water on the surface is important in the oxidation of iron through the formation of hydroxyl species. Unless your vacuum is an ultra high vacuum system and has been baked, you will always have water absorbed on the surface. A desiccator that has only been rough pumped or (heaven forbid) pumped using a house vacuum has plenty of both oxygen and water available for oxidation. We have introduced a new product called the SampleSaver(TM) Storage Container that will help in the prevention of oxidation of samples. This unit uses an inert gas such as Ar, N2, or CO2 to purge the volume of the container and then to compress the inert gas slightly so to prevent diffusion into the container. I have used this successfully with XTEM samples that readily oxidized when made, but I have not tried it with Fe samples. We have a holder specially designed for TEM samples. Contact me offline to discuss your problem.
3) There are a few things that can help with beam sensitive samples. When I was with PPG, I had some problems with glass samples. First, what accelerating voltage are you using? Increasing the voltage will decrease the deposition of energy into the sample and will help stop beam damage. 200 keV should be considered the minimum that you should use. For the same reason, thinner samples will help. When I was forced to use a 120 keV machine for the glass samples, I had some success with putting a thin carbon coating on the two surfaces. I used both evaporation and ion sputtering to put the carbon down. It is important to put the right amount down, too much interferes too much with your imaging and too little doesn't do much to help. I modified the configuration of my ion mill for ion depositing the carbon by sputtering. Alternatively, you could use a commercially available ion sputter deposition system for high resolution SEM such as our IBS/e system for putting a controlled uniform layer of carbon on the sample.
Disclaimer: South Bay Technology (SBT), Inc. is the representative for Technoorg-Linde products in the United States. SBT manufactures and sells the MicroCleave(TM) kit, SampleSaver(TM) Storage Container, and the IBS/e ion beam sputter deposition and etching system.
-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
Sent: Monday, April 03, 2006 7:20 PM To: Walck-at-SouthBayTech.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both chao.wang-at-materials.ox.ac.uk as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: chao.wang-at-materials.ox.ac.uk Name: Chao Wang
Organization: Oxford Materials
Title-Subject: [Filtered] help for MgO prepration and Fe oxidation
Question: Dear All
I have three problems, could you tell me some suggestions:
1.single crystale MgO preparation
I need very good quality cross section TEM sample. I now manully polish it to 70 micron and dimple to abount 20 micron and then ion milling (PIPS). But it's not so good. The edge looks sharp and MgO cracks a lot when I ground lower then 60 micron on diamond papers. Do you have better ideas to get very thin specimen?
2. Fe oxidiation Another problem is Fe oxidation (on top of MgO substrate), it seems after some time, the sample oxidized a lot, even I got very thin specimen, it looks like amorphous (it should be crystalline). How to keep it? (I use plasma cleaning, not working very well)
3. Beam sensitive to MgO sample
Sometime the beam in TEM is sensitive to MgO sample, how to get rid of this?
I am looking for a small (lunch box size) vacuum container. This is for transporting SEM specimens under vacuum to avoid oxidation between sites.
I will be using a small diaphram or rotary pump with KF15 fitting. The unit does not have to hold initial vacuum for a long time. Plastic or metal is not a key factor but durability is in construction and long term use.
Any ideas?
Vendor input is welcome.
gary g.
==============================Original Headers============================== 7, 17 -- From gary-at-gaugler.com Tue Apr 4 18:17:49 2006 7, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k34NHlmV017348 7, 17 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 18:17:48 -0500 7, 17 -- Received: (qmail 8197 invoked from network); 4 Apr 2006 16:16:48 -0700 7, 17 -- Received: by simscan 1.1.0 ppid: 8194, pid: 8195, t: 0.1041s 7, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 7, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 7, 17 -- by qsmtp2 with SMTP; 4 Apr 2006 16:16:47 -0700 7, 17 -- Message-Id: {7.0.1.0.2.20060404161341.023138a0-at-gaugler.com} 7, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 7, 17 -- Date: Tue, 04 Apr 2006 16:17:43 -0700 7, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 7, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 7, 17 -- Subject: Small vacuum box 7, 17 -- Mime-Version: 1.0 7, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Gary, I have tested our new SampleSaver(TM) Storage Container under vacuum with a diaphragm pump and it holds a vacuum quite well for a number of days (length of test). However, I have not tested it whether it will protect samples from oxidation as well as purging the unit with N2 and I have also not checked the pressure change during that time. We have a number of inserts for the unit that will hold 1/8" pin SEM stubs and 3/8" SEM stubs as well as one that will hold three TEM grid boxes. There is a insert unit available that will hold a 1-1/4" metallurgical sized sample. The different SEM sample plates can be mixed and matched. We have a larger unit that was designed for holding FIB Lift-out samples for storage and transport that will also accept the 1/8" pins. The units are light and perfect for transporting samples from site to site.
The advantage that I found for purging is that there is not always a vacuum pump available at the other end of your trip when transporting samples. In fact, there isn't always a spare pure Ar or N2 gas line available either. That's why we also designed the Thing-A-Ma-Jug(TM) gas supply system that will purge the unit with the head space from liquid nitrogen which almost always available in the typical EM lab and is also a source of pure N2. The Thing-A-Ma-Jug(TM) is also light enough to be very portable. The SampleSaver(TM) is designed to pressurize the container when the purging is done and thus inhibit gas diffusion through the plastic container and O-rings.
If you have any questions on the system, please give me a call.
Disclaimer: SBT makes and sells the SampleSaver(TM) storage container system and Thing-A-Ma-Jug(TM) gas supply system.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com] Sent: Tuesday, April 04, 2006 4:25 PM To: Walck-at-SouthBayTech.com
Listers:
I am looking for a small (lunch box size) vacuum container. This is for transporting SEM specimens under vacuum to avoid oxidation between sites.
I will be using a small diaphram or rotary pump with KF15 fitting. The unit does not have to hold initial vacuum for a long time. Plastic or metal is not a key factor but durability is in construction and long term use.
Any ideas?
Vendor input is welcome.
gary g.
==============================Original Headers============================== 7, 17 -- From gary-at-gaugler.com Tue Apr 4 18:17:49 2006 7, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k34NHlmV017348 7, 17 -- for {microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 18:17:48 -0500 7, 17 -- Received: (qmail 8197 invoked from network); 4 Apr 2006 16:16:48 -0700 7, 17 -- Received: by simscan 1.1.0 ppid: 8194, pid: 8195, t: 0.1041s 7, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 7, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 7, 17 -- by qsmtp2 with SMTP; 4 Apr 2006 16:16:47 -0700 7, 17 -- Message-Id: {7.0.1.0.2.20060404161341.023138a0-at-gaugler.com} 7, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 7, 17 -- Date: Tue, 04 Apr 2006 16:17:43 -0700 7, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 7, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 7, 17 -- Subject: Small vacuum box 7, 17 -- Mime-Version: 1.0 7, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 20, 27 -- From walck-at-southbaytech.com Tue Apr 4 19:56:24 2006 20, 27 -- Received: from ylpvm01.prodigy.net (ylpvm01-ext.prodigy.net [207.115.57.32]) 20, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k350uOEU028572 20, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 19:56:24 -0500 20, 27 -- Received: from pimout6-ext.prodigy.net (pimout6-int.prodigy.net [207.115.4.22]) 20, 27 -- by ylpvm01.prodigy.net (8.12.10 outbound/8.12.10) with ESMTP id k350uLai022661 20, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Apr 2006 20:56:21 -0400 20, 27 -- X-ORBL: [64.169.193.90] 20, 27 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 20, 27 -- by pimout6-ext.prodigy.net (8.13.4 outbound domainkey aix/8.13.4) with ESMTP id k350uH66091756; 20, 27 -- Tue, 4 Apr 2006 20:56:22 -0400 20, 27 -- From: "Scott Walck" {walck-at-southbaytech.com} 20, 27 -- To: {gary-at-gaugler.com} 20, 27 -- Cc: {Microscopy-at-microscopy.com} 20, 27 -- Subject: RE: [Microscopy] Small vacuum box 20, 27 -- Date: Tue, 4 Apr 2006 17:56:23 -0700 20, 27 -- Message-ID: {006201c6584b$c5fe59d0$7801a8c0-at-dynamicbl8uno3} 20, 27 -- MIME-Version: 1.0 20, 27 -- Content-Type: text/plain; 20, 27 -- charset="us-ascii" 20, 27 -- Content-Transfer-Encoding: 7bit 20, 27 -- X-Priority: 3 (Normal) 20, 27 -- X-MSMail-Priority: Normal 20, 27 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 20, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 20, 27 -- In-Reply-To: {200604042324.k34NOWEc023825-at-ns.microscopy.com} 20, 27 -- Importance: Normal ==============================End of - Headers==============================
Have a look at a vacuum components catalog, like Lesker, MDC, EVAC or so. I have used KF40 or ISO-K pipes as container. You may find PVC, Al alloy, or SS fittings. Durability is garanteed ! Can be cleaned with solvants, baked for degasing, and some workshops can modify standard components for cheap (directe weld a vaccum valve, for exemple). I have a old zeolite foreline trap, which I will use next as container.
Hope it helps
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 9, 30 -- From jacques.faerber-at-ipcms.u-strasbg.fr Wed Apr 5 04:47:45 2006 9, 30 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.154]) 9, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k359liAU013151 9, 30 -- for {microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 04:47:45 -0500 9, 30 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 9, 30 -- by mailhost.u-strasbg.fr (8.13.4/jtpda-5.5pre1) with ESMTP id k359lfWJ025271 9, 30 -- ; Wed, 5 Apr 2006 11:47:41 +0200 (CEST) 9, 30 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr [130.79.54.3]) 9, 30 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id ED02F1000126; 9, 30 -- Wed, 5 Apr 2006 11:47:31 +0200 (CEST) 9, 30 -- Message-ID: {4433923F.3000501-at-ipcms.u-strasbg.fr} 9, 30 -- Date: Wed, 05 Apr 2006 11:47:43 +0200 9, 30 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 9, 30 -- User-Agent: Mozilla Thunderbird 1.0.7 (X11/20051011) 9, 30 -- X-Accept-Language: fr, en 9, 30 -- MIME-Version: 1.0 9, 30 -- To: gary-at-gaugler.com, Microscopy-at-microscopy.com 9, 30 -- Subject: Re: [Microscopy] Small vacuum box 9, 30 -- References: {200604042325.k34NPd5P025606-at-ns.microscopy.com} 9, 30 -- In-Reply-To: {200604042325.k34NPd5P025606-at-ns.microscopy.com} 9, 30 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 30 -- Content-Transfer-Encoding: 8bit 9, 30 -- X-IPCMS-MailScanner: Found to be clean 9, 30 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 9, 30 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.154]); Wed, 05 Apr 2006 11:47:41 +0200 (CEST) 9, 30 -- X-Virus-Scanned: ClamAV 0.88/1376/Wed Apr 5 07:51:25 2006 on mr4.u-strasbg.fr 9, 30 -- X-Virus-Status: Clean 9, 30 -- X-Spam-Status: No, score=-1.4 required=5.0 tests=ALL_TRUSTED,AWL 9, 30 -- autolearn=disabled version=3.1.0 9, 30 -- X-Spam-Checker-Version: SpamAssassin 3.1.0 (2005-09-13) on mr4.u-strasbg.fr ==============================End of - Headers==============================
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Good Morning, Rhonda: } } We have a Leica EM-Stain. We purchased it about a year ago in a } quest to make our staining absolutely consistent, and in regard to that, } it works wonderfully. Given all of the options for wetting times, stain } times, stain temperatures, etc, it's pretty versatile and dead-on } accurate, not to mention it frees up a lot of a technician's time! } However, we're had nothing but problems with the unit. We've gone } though 3 pumps already, numerous service calls, etc. In Leica's defense, } they may have had a bad batch of stain (high amounts of precipitate), } which is clogging up the pump and the filters. Speaking of the stain, } you're "required" to use their pre-made stains, else you void the warranty } on the unit. There's a bit of upkeep with the stainer as well, in that you } must perform a cleaning cycle at least once a week (which only takes a few } minutes to get running but someone still has to remember to do it), and it } uses 3% Nitric acid for the cleaning cycle, which possibly adds another } waste stream for your lab. } All in all, it would be a great unit if there weren't so many } problems with it. We may just have ended up with a lemon, though. Hard to } say. } } -Chris } } } } } } } rra-at-stowers-institute.org } 04/03/2006 10:20 PM } Please respond to rra } } } To: Christopher Hayden/PH/Novartis-at-PH } cc: } Subject: [Microscopy] viaWWW: Leica EM-Stain } } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both rra-at-stowers-institute.org as well as the MIcroscopy } Listserver } --------------------------------------------------------------------------- } } 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 } } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 7, 12 -- From zaluzec-at-microscopy.com Mon Apr 3 21:12:03 2006 } 7, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 7, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id k342C2tI026972 } 7, 12 -- for {microscopy-at-microscopy.com} ; Mon, 3 Apr 2006 } 21:12:02 -0500 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: (Unverified) } 7, 12 -- Message-Id: {p06110403c0578663217d-at-[206.69.208.22]} } 7, 12 -- Date: Mon, 3 Apr 2006 21:12:01 -0500 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: rra-at-stowers-institute.org (by way of MicroscopyListserver) } 7, 12 -- Subject: viaWWW: Leica EM-Stain } 7, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== } } } } } ==============================Original Headers============================== } 25, 30 -- From christopher.hayden-at-novartis.com Tue Apr 4 06:11:07 2006 } 25, 30 -- Received: from mail84.messagelabs.com (mail84.messagelabs.com [195.245.231.99]) } 25, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k34BB6Kf017329 } 25, 30 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 4 Apr 2006 06:11:06 -0500 } 25, 30 -- X-VirusChecked: Checked } 25, 30 -- X-Env-Sender: christopher.hayden-at-novartis.com } 25, 30 -- X-Msg-Ref: server-4.tower-84.messagelabs.com!1144149064!19910770!1 } 25, 30 -- X-StarScan-Version: 5.5.9.1; banners=-,-,- } 25, 30 -- X-Originating-IP: [160.62.1.174] } 25, 30 -- Received: (qmail 13192 invoked from network); 4 Apr 2006 11:11:05 -0000 } 25, 30 -- Received: from ch2ssaenov01.novartis.com (HELO ch2ssaenov01.novartis.com) (160.62.1.174) } 25, 30 -- by server-4.tower-84.messagelabs.com with AES256-SHA encrypted SMTP; 4 Apr 2006 11:11:05 -0000 } 25, 30 -- Received: from mtap2.is.chbs ([192.37.33.19]) } 25, 30 -- by ch2ssaenov01.novartis.com (8.13.6/8.13.4) with ESMTP id k34B8vkO027268; } 25, 30 -- Tue, 4 Apr 2006 13:08:57 +0200 } 25, 30 -- Received: from phchbs-s3025.EU.novartis.net (phchbs-s3025.eu.novartis.net [192.37.31.249]) } 25, 30 -- by mtap2.is.chbs (Switch-3.1.6/Switch-3.1.6) with ESMTP id k34BB3nH9486452; } 25, 30 -- Tue, 4 Apr 2006 13:11:04 +0200 } 25, 30 -- To: Microscopy-at-msa.microscopy.com } 25, 30 -- Cc: rra-at-stowers-institute.org } 25, 30 -- Subject: Re: [Microscopy] viaWWW: Leica EM-Stain } 25, 30 -- MIME-Version: 1.0 } 25, 30 -- X-Mailer: Lotus Notes Release 5.0.12 February 13, 2003 } 25, 30 -- Message-ID: {OF829E7CC2.942F7902-ON85257146.003C549D-85257146.003D705C-at-EU.novartis.net} } 25, 30 -- From: christopher.hayden-at-novartis.com } 25, 30 -- Date: Tue, 4 Apr 2006 07:11:20 -0400 } 25, 30 -- X-MIMETrack: Serialize by Router on CHBSSPH0/PH/Novartis(5012HF433 | October 14, 2003) at } 25, 30 -- 04.04.2006 13:11:04, } 25, 30 -- Serialize complete at 04.04.2006 13:11:04 } 25, 30 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
-- Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Zentrum für Molekulare Medizin Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both a.d.mckinnon-at-abdn.ac.uk as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both xiuhuixin-at-yahoo.com.cn as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: xiuhuixin-at-yahoo.com.cn Name: Huixin
Title-Subject: [Filtered] Help for FIB prepared specimens
Question: The first problem I met is that sometimes I lost my specimens. The machine I used is a single beam FIB and the material is GaN. After ex-situ lift-out, I put the specimens onto a copper mesh. I used membrane box to store the specimens. However, when I load the specimen into the TEM specimen holder, I cannot find the specimen in the microscope sometimes. Does anybody have a method to prevent specimens from being lost?
The second problem I met is that there is too much surface damage on my specimens. Does anybody have some tips to reduce the surface damage? Thanks all.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both marie.cheynet-at-ltpcm.inpg.fr as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: marie.cheynet-at-ltpcm.inpg.fr Name: Cheynet Marie
Organization: CNRS-France
Title-Subject: [Filtered] polycrystalline alumina cap
Question: Bonjour,
I am looking for a solvent to dissolve a 10 nm thick polycrystalline alumina cap deposited on a ultra-thin HfO2 dielectric layer. Thanks for your suggestions. marie
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both mmiralles-at-pi.ac.ae as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mmiralles-at-pi.ac.ae Name: myleen
Title-Subject: [Filtered] E-SEM Buy-off Checklist
Question: hi,
we will be acquiring an E-SEM very soon (due for delivery this May 2006). just wondering if anyone has an equipment buy-off checklist i could use as a pattern for checking the machine?
Please reply to the list as well -- I have the same instrument, and the same question.
Phil
} This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both a.d.mckinnon-at-abdn.ac.uk as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: a.d.mckinnon-at-abdn.ac.uk } Name: Alastair McKinnon } } Organization: University of Aberdeen } } Title-Subject: [Filtered] Histology & ElectronMicroscopy } } Question: Can anyone suggest an effective algicide that is safe to } use in a chiller circulator supplying water at 18-20'C for a Philips } CM10 TEM. -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 3, 22 -- From oshel1pe-at-cmich.edu Wed Apr 5 12:11:38 2006 3, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35HBcAD013817 3, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 12:11:38 -0500 3, 22 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 3, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k35Hn54l015106 3, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 13:49:05 -0400 3, 22 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 3, 22 -- Wed, 5 Apr 2006 13:11:36 -0400 3, 22 -- Mime-Version: 1.0 3, 22 -- Message-Id: {f0623090ac059aa9c0b5f-at-[141.209.160.132]} 3, 22 -- In-Reply-To: {200604051705.k35H5f8q006805-at-ns.microscopy.com} 3, 22 -- References: {200604051705.k35H5f8q006805-at-ns.microscopy.com} 3, 22 -- Date: Wed, 5 Apr 2006 13:11:33 -0400 3, 22 -- To: Microscopy-at-microscopy.com 3, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 22 -- Subject: Re: [Microscopy] viaWWW: Histology & ElectronMicroscopy 3, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 22 -- X-OriginalArrivalTime: 05 Apr 2006 17:11:36.0758 (UTC) FILETIME=[FF157D60:01C658D3] 3, 22 -- X-CanItPRO-Stream: default 3, 22 -- X-Spam-Score: -4 () L_EXCH_MF 3, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
On Apr 5, 2006, at 9:56 AM, a.d.mckinnon-at-abdn.ac.uk wrote:
} Question: Can anyone suggest an effective algicide that is safe to use } in a chiller circulator supplying water at 18-20'C for a Philips CM10 } TEM. } Dear Alastair, We sprinkle a small amount of 2,2'-Methylenebis(4-chlorophenol)--dichlorophene for short--onto the surface of the chiller water. It is not very soluble, so it floats on the surface in clumps. When we don't see any more clumps, we add a little more dichlorophene. I also used this compound in the chillers on the HVEM in Albany NY for more than 20 years, and there was no observable harm to either the chillers or the scope; I measured the water flow rates through the lenses annually, and there was essentially no change over the entire time. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Apr 5 13:09:36 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35I9a0Y024296 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 5 Apr 2006 13:09:36 -0500 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP 4, 22 -- id 7E40B3496E; Wed, 5 Apr 2006 11:09:35 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP 4, 22 -- id 2F7EF34A05; Wed, 5 Apr 2006 11:09:34 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200604051656.k35Gu7ik007931-at-ns.microscopy.com} 4, 22 -- References: {200604051656.k35Gu7ik007931-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {ea030c84e0b3f694978483884f24db9d-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: Histology & ElectronMicroscopy 4, 22 -- Date: Wed, 5 Apr 2006 11:19:02 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com, a.d.mckinnon-at-abdn.ac.uk 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
A colleage is looking for a red fluorescent dye to stain lignin (or plant cell wall) for viewing in a confocal microscope. Any suggestions would be appreciated.
Thank you.
JB -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 3, 16 -- From bozzola-at-siu.edu Wed Apr 5 13:10:17 2006 3, 16 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35IAHpR025376 3, 16 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 5 Apr 2006 13:10:17 -0500 3, 16 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 3, 16 -- by abbmta2.siu.edu (Switch-3.1.8/Switch-3.1.6) with ESMTP id k35IAHpT011183 3, 16 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 5 Apr 2006 13:10:17 -0500 (CDT) 3, 16 -- Mime-Version: 1.0 3, 16 -- X-Sender: bozzola-at-saluki-mail.siu.edu 3, 16 -- Message-Id: {p06110408c059b8174ef1-at-[131.230.177.142]} 3, 16 -- Date: Wed, 5 Apr 2006 13:10:15 -0500 3, 16 -- To: Microscopy-at-msa.microscopy.com 3, 16 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 3, 16 -- Subject: LM: flouresc cw stain 3, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 16 -- X-MASF: 0.00% ==============================End of - Headers==============================
On Apr 5, 2006, at 9:57 AM, marie.cheynet-at-ltpcm.inpg.fr wrote:
} I am looking for a solvent to dissolve a 10 nm thick polycrystalline } alumina cap deposited on a ultra-thin HfO2 dielectric layer. } Thanks for your suggestions. } Dear Marie, I do not know what the effect would be on the HfO2, but Al2O3 will dissolve in strong base. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Apr 5 13:12:00 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35IBxMb030897 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 5 Apr 2006 13:12:00 -0500 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP 4, 22 -- id A5C0B10ACB5; Wed, 5 Apr 2006 11:11:59 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP 4, 22 -- id B9D0810AC49; Wed, 5 Apr 2006 11:11:58 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200604051657.k35Gv9OF009943-at-ns.microscopy.com} 4, 22 -- References: {200604051657.k35Gv9OF009943-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {7e0081e0eed22f808166d15dd8fd0c14-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: polycrystalline alumina cap 4, 22 -- Date: Wed, 5 Apr 2006 11:21:25 -0700 4, 22 -- To: marie.cheynet-at-ltpcm.inpg.fr, microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Congo Red binds cellulose and would therefore be a good fluorescence stain for plant cell walls emitting in the red (emission~ 590 nm). As for a fluorescent lignin stain, keep in mind that lignin is autofluorescent (broadband emission ~470-520 nm). It could be stained with a fluorescent Schiff's base (e.g., Auramine O) as well.
Hope this helps.
Howard
On Apr 5, 2006, at 1:11 PM, bozzola-at-siu.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 } ---------------------------------------------------------------------- } ------ } } A colleage is looking for a red fluorescent dye to stain lignin (or } plant cell wall) for viewing in a confocal microscope. Any } suggestions would be appreciated. } } Thank you. } } JB } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/~image/ } ############################################################## } } ==============================Original } Headers============================== } 3, 16 -- From bozzola-at-siu.edu Wed Apr 5 13:10:17 2006 } 3, 16 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu } [131.230.254.206]) } 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k35IAHpR025376 } 3, 16 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 5 Apr 2006 } 13:10:17 -0500 } 3, 16 -- Received: from [131.230.177.142] } (ws177142.microscope.siu.edu [131.230.177.142]) } 3, 16 -- by abbmta2.siu.edu (Switch-3.1.8/Switch-3.1.6) with ESMTP } id k35IAHpT011183 } 3, 16 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 5 Apr 2006 } 13:10:17 -0500 (CDT) } 3, 16 -- Mime-Version: 1.0 } 3, 16 -- X-Sender: bozzola-at-saluki-mail.siu.edu } 3, 16 -- Message-Id: {p06110408c059b8174ef1-at-[131.230.177.142]} } 3, 16 -- Date: Wed, 5 Apr 2006 13:10:15 -0500 } 3, 16 -- To: Microscopy-at-msa.microscopy.com } 3, 16 -- From: "John J. Bozzola" {bozzola-at-siu.edu} } 3, 16 -- Subject: LM: flouresc cw stain } 3, 16 -- Content-Type: text/plain; charset="us-ascii" ; } format="flowed" } 3, 16 -- X-MASF: 0.00% } ==============================End of - } Headers==============================
R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility Danforth Plant Science Center 975 N. Warson Rd. St. Louis, MO 63132
ph 314-587-1261 fx 314-587-1361 cell 314-378-2409 rhberg-at-danforthcenter.org www.danforthcenter.org visit this educational resource: http://www.danforthcenter.org/Cells/
==============================Original Headers============================== 12, 23 -- From RHBerg-at-danforthcenter.org Wed Apr 5 14:14:55 2006 12, 23 -- Received: from spm1.ddpsc.org (spm1.danforthcenter.org [65.254.111.26]) 12, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35JEs5w021491 12, 23 -- for {microscopy-at-sparc5.microscopy.com} ; Wed, 5 Apr 2006 14:14:54 -0500 12, 23 -- Received: from spm1.ddpsc.org (127.0.0.1) by spm1.ddpsc.org (MlfMTA v3.1r24) id h6gbis0171sp for {microscopy-at-sparc5.microscopy.com} ; Wed, 5 Apr 2006 14:14:53 -0500 (envelope-from {RHBerg-at-danforthcenter.org} ) 12, 23 -- Received: from mail02.ddpsc.org ([10.101.0.23]) 12, 23 -- by spm1.ddpsc.org (MailFrontier 4.5.7.7473) 12, 23 -- with ESMTP; Wed, 05 Apr 2006 14:14:53 -0500 12, 23 -- Received: from [10.14.0.20] ([10.14.0.20] unverified) by mail02.ddpsc.org with Microsoft SMTPSVC(5.0.2195.6713); 12, 23 -- Wed, 5 Apr 2006 14:14:53 -0500 12, 23 -- Mime-Version: 1.0 (Apple Message framework v746.3) 12, 23 -- In-Reply-To: {200604051811.k35IBiYY030143-at-ns.microscopy.com} 12, 23 -- References: {200604051811.k35IBiYY030143-at-ns.microscopy.com} 12, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 23 -- Message-Id: {6DB37DB7-A5C8-4D9D-BED7-62E6916A3907-at-danforthcenter.org} 12, 23 -- Content-Transfer-Encoding: 7bit 12, 23 -- From: "R. Howard Berg" {rhberg-at-danforthcenter.org} 12, 23 -- Subject: Re: [Microscopy] LM: flouresc cw stain 12, 23 -- Date: Wed, 5 Apr 2006 14:14:50 -0500 12, 23 -- To: microscopy-at-ns.microscopy.com 12, 23 -- X-Mailer: Apple Mail (2.746.3) 12, 23 -- X-OriginalArrivalTime: 05 Apr 2006 19:14:53.0007 (UTC) FILETIME=[379785F0:01C658E5] 12, 23 -- X-Mlf-Version: 4.5.7.7473 ==============================End of - Headers==============================
Concerning Gary's inquiry about a small vacuum chamber for specimen transfer.
I have produced specimen rods for two models of TEMs that give protection of a specimen from the atmosphere for a brief period while being moved from a reaction chamber into the microscope specimen stage, and also a small glass reaction chamber for use with such holders - if such might be of interest in this type of application. The glass reactor is small enough to be used for the kind of operation mentioned. -- 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:734-662-5237 Address mail to: 1136 Mixtwood Rd. Ann Arbor, MI 48103-3035
==============================Original Headers============================== 2, 14 -- From bigelow-at-engin.umich.edu Wed Apr 5 14:55:39 2006 2, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 2, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35JtdTb031483 2, 14 -- for {microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 14:55:39 -0500 2, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 2, 14 -- by srvr22.engin.umich.edu (8.13.6/8.13.5) with ESMTP id k35JtcGY026563 2, 14 -- for {microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 15:55:38 -0400 (EDT) 2, 14 -- Mime-Version: 1.0 2, 14 -- Message-Id: {p06210202c059cf72df14-at-[141.212.131.221]} 2, 14 -- Date: Wed, 5 Apr 2006 15:55:37 -0400 2, 14 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 2, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 2, 14 -- Subject: [Microscopy] Specimen protection fro atmosphere 2, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Would anyone have a detailed protocol for preparing mouse sp*rm suspensions for TEM that they would be willing to share? I've been attempting to concentrate my sample by centrifugation, then resuspend in agar, but I'm not getting very good results.
Many thanks in advance for any suggestions, comments, or protocols.
(Note: I seem to be having some trouble posting due to my subject matter. Please forgive the work-around!)
Best regards,
Angela
-- Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West and 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-796-5977 Fax: 212-496-3480
==============================Original Headers============================== 8, 29 -- From avklaus-at-amnh.org Wed Apr 5 15:48:54 2006 8, 29 -- Received: from lepore.amnh.org (lepore.amnh.org [216.73.241.12]) 8, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35KmsTM009271 8, 29 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 15:48:54 -0500 8, 29 -- Received: from localhost (localhost [127.0.0.1]) 8, 29 -- by lepore.amnh.org (Postfix) with ESMTP id D237F5B7A1; 8, 29 -- Wed, 5 Apr 2006 16:48:53 -0400 (EDT) 8, 29 -- Received: from lepore.amnh.org ([127.0.0.1]) 8, 29 -- by localhost (lepore.amnh.org [127.0.0.1]) (amavisd-new, port 10024) 8, 29 -- with ESMTP id 01224-04; Wed, 5 Apr 2006 16:48:52 -0400 (EDT) 8, 29 -- Received: from webmail.amnh.org (cain.amnh.org [216.73.241.17]) 8, 29 -- by lepore.amnh.org (Postfix) with ESMTP id C760B5B795; 8, 29 -- Wed, 5 Apr 2006 16:48:52 -0400 (EDT) 8, 29 -- Received: from 216.73.250.195 8, 29 -- (SquirrelMail authenticated user avklaus) 8, 29 -- by webmail.amnh.org with HTTP; 8, 29 -- Wed, 5 Apr 2006 16:48:52 -0400 (EDT) 8, 29 -- Message-ID: {2020.216.73.250.195.1144270132.squirrel-at-webmail.amnh.org} 8, 29 -- Date: Wed, 5 Apr 2006 16:48:52 -0400 (EDT) 8, 29 -- Subject: TEM protocol: male gametes 8, 29 -- From: "Angela V. Klaus" {avklaus-at-amnh.org} 8, 29 -- To: Microscopy-at-microscopy.com 8, 29 -- User-Agent: SquirrelMail/1.4.4 8, 29 -- MIME-Version: 1.0 8, 29 -- Content-Type: text/plain;charset=iso-8859-1 8, 29 -- Content-Transfer-Encoding: 8bit 8, 29 -- X-Priority: 3 (Normal) 8, 29 -- Importance: Normal 8, 29 -- X-Virus-Scanned: amavisd-new at amnh.org ==============================End of - Headers==============================
Electron Microscopist / Lab Manager The Catholic University of America, Vitreous State Laboratory, Washington, D.C.
Appointment Date/Term: Immediate/permanent
Salary and Benefits: Negotiable, 2x matching 403b, (up to 5% of salary after 1 year,) 21 days paid vacation, 15 days paid holiday, sick leave, health and life insurance, flexible spending account for medical/dental reimbursement, tuition reimbursement, (up to 6 credits per semester,) and relocation assistance negotiable.
Essential Duties: Microstructural characterization of materials utilizing OLM, SEM, EDS, WDS, TEM, STEM, and XRD. Provide technical expertise to researchers on equipment operation and microscopy laboratory protocols. Conduct basic EM maintenance on related equipment.
Qualifications: Candidate must either have a two-year degree in electron microscopy or a BS in physics, chemistry, or materials science with 3-5 years of hands-on electron microscopy experience. Candidate must have detailed operational knowledge of scanning and transmission electron microscopes, as well as, energy-dispersive spectroscopy systems, fundamental understanding of image processing and analysis techniques, capacity to prepare SEM specimens utilizing standard metallographic techniques, ability to prepare TEM specimens via tripod polishing, jet polishing, dimpling/ion milling, and extraction replication, a generally good understanding of electron microscopy lab maintenance, and strong verbal and written communication skills. EM lab management experience a major plus.
Instrumentation: JEOL 5900LV SEM with Oxford INCA ENERGY 300/WAVE 700 EDS/WDS systems, JEOL 35c SEM with two JEOL FCS WD-spectrometers interfaced to Noran Vantage EDS system, JEOL 2000EX / FX TEM/STEM w/Oxford INCA-ISIS ENERGY 200 EDS, Olympus upright light microscope w/ brightfield transmitted and reflected/polarizing light, Leica inverted stage w/ brightfield, darkfield, polarizing, Nomarski DIC.
Cavin T. F. Mooers EM Facility Manager The Catholic University of America Vitreous State Laboratory Hannan Hall Rm 433 Washington, D.C. 20064 (202) 319-6237 (Office) (202) 319-5346 (Lab) (202) 319-4469 (Fax)
Send resume and cover letter with salary requirements via email reply.
______________ ______________ ______________ ______________ Sent via VSL Webmail
==============================Original Headers============================== 12, 19 -- From CavinM-at-vsl.cua.edu Wed Apr 5 16:30:08 2006 12, 19 -- Received: from hermes.vsl.cua.edu (interface.vsl.cua.edu [136.242.188.2]) 12, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k35LU4ZW019652 12, 19 -- for {microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 16:30:05 -0500 12, 19 -- X-Filtered-With-Copfilter: Version 0.82 (ProxSMTP 1.3.91) 12, 19 -- X-Copfilter-Virus-Scanned: ClamAV 0.88/1376 - Wed Apr 5 00:51:25 2006 12, 19 -- X-Copfilter: Client is part of our network, skipped SpamAssassin 12, 19 -- Received: from 136.242.189.141 with HTTP 12, 19 -- by webserver hermes.vsl.cua.edu (136.242.189.3) ; Wed, 5 Apr 2006 17:30:03 EDT 12, 19 -- Date: Wed, 5 Apr 2006 17:30:04 -0400 12, 19 -- Message-Id: {200604051730.AA171114822-at-hermes.vsl.cua.edu} 12, 19 -- Mime-Version: 1.0 12, 19 -- Content-Type: text/plain; charset=us-ascii 12, 19 -- From: "Cavin Mooers" {CavinM-at-vsl.cua.edu} 12, 19 -- Reply-To: {CavinM-at-vsl.cua.edu} 12, 19 -- X-Sender: {CavinM-at-vsl.cua.edu} 12, 19 -- To: {microscopy-at-microscopy.com} 12, 19 -- Subject: Position available: Electron Microscopist/Lab Manager 12, 19 -- X-Mailer: {IMail v8.21} ==============================End of - Headers==============================
Thanks to all for the suggestions. I suppose that I should have provided more info about what I seek.
I am looking for a small vacuum container that would hold specimens in storage containers like Pella 16708 which have plastic specimen locations like Pella 16166.
The plastic box is 3-11/16 x 2-11/16 x 1-1/2" and will not fit in a KF25 pipe. So I need some little dessicator unit that is easily transportable. The issue is getting the specimen out of RIE and/or FIB tool over to EBSD tool without growing too much oxide such that EBSD does not pattern.
gary g.
At 04:21 PM 4/4/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
FIB Lift-Out Protection: At the MRS meeting in San Francisco (April 17-21), we are introducing the large size SampleSaver(TM) Storage Container (model# SS-200) along with FIB specimen holders with CastleGuard(TM) protection. These holders are designed to hold a cut grid or an Omniprobe(R) grid in the upright position for the in-situ lift-out process in the FIB. After removal from the FIB, it can be stored in a numbered insert for the SS-200 and held in place with a set screw so that the SampleSaver(TM) unit can be transported. I have placed OmniProbe grids into these holders and dropped them repeatedly from a height of six feet without any of them falling out. The CastleGuard protection prevents damage to the sample even if dropped to the floor head-first, i.e. specimen side down. We are about to send some samples around the world to Hungary to see if they can survive FedEx international handling. (Anybody want to take bets?) If you are interested in this sample holder, I can send an image if you respond to me offline.
FIB Damage: To remove the surface damage from FIB samples, you need to ion mill at low energies at low angles. We sell the Gentle Mill which was specifically designed to remove the surface layer from FIB samples and conventional ion milling samples for high resolution applications. Our IV3/IV4 ion mill system can also be configured with a low energy gun. This gun, designed and patented by Arpa Barna and associates, can operate at an accelerating voltage from as low as 100 eV up to 2 kV. It uses an electron source to increase ionization of the gas and an Einzel lens to focus the ions at the sample to increase ion current density. At 2 kV, this gun can rival the thinning rates of other guns operating around 5 kV. If you visit our application notes section of our website, we have an application note entitled, "Applications of the GentleMill(TM) To FIB Prepared TEM Samples" that shows what it can do on an FIB prepared TEM sample. Barna and Pecz (A. Barna and B. Pecz, Ultramicroscopy 70, 1998) have shown that conventional ion milling with 3 keV Ar ions can give an amorphous layer of as much as 120 A on each ion milled surface, but with 250 eV the layer is only about 10 A. I can also send you the data for that paper if you contact me offline.
Disclaimer: We did not "Plant" these questions!! South Bay Technology, Inc. manufactures and sells the SampleSaver(TM) storage container and the FIB specimen holder with CastleGuard(TM) protection and sells the Technoorg-Linde Gentle Mill(TM) and IV3/IV4 ion mills.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: xiuhuixin-at-yahoo.com.cn [mailto:xiuhuixin-at-yahoo.com.cn] Sent: Wednesday, April 05, 2006 10:04 AM To: Walck-at-SouthBayTech.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both xiuhuixin-at-yahoo.com.cn as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: xiuhuixin-at-yahoo.com.cn Name: Huixin
Title-Subject: [Filtered] Help for FIB prepared specimens
Question: The first problem I met is that sometimes I lost my specimens. The machine I used is a single beam FIB and the material is GaN. After ex-situ lift-out, I put the specimens onto a copper mesh. I used membrane box to store the specimens. However, when I load the specimen into the TEM specimen holder, I cannot find the specimen in the microscope sometimes. Does anybody have a method to prevent specimens from being lost?
The second problem I met is that there is too much surface damage on my specimens. Does anybody have some tips to reduce the surface damage? Thanks all.
I do not think it hard to prepare the cross-sectional MgO samples. What you need is the patience. Gatan kits are very convenient!
1) Prepare the sandwitch, and glue the slabs face-to-face. The M-bond is good enough, but G-1 epoxy from Gatan is better for MgO samples.
2) You should polish the 1st side very well. I usually use the 5micron sand paper before I polish the sandwitch with 1 micron diamond paste. Just use the big polishing machine which can be found almost each metallurgy lab. Since MgO is very easy to be polished, you can directly go to 1 micron after the grinding with sand paper. Please check with light microscope at 200x, you should find no scratches.
3) After you turn to 2nd side, first grind the sandwitch to about 100-80 microns. 5-micron sand paper should be the used finally. Then you can mount the samples on dimpler, and dimpler the disk down to 35 microns (3-micron diamond paste). You may see some small scratches at the center due to the dimpling with metal wheels.
4) The you can polish the samples down to 15 microns thick on the dimpler machines. I usually use the large force and high speed. First try 5 or 3 microns diamond paste, and then 1 micron finally. It usually takes more than half hour to get the good samples. But it deserves!
5) when you glue the disk on the Cu-grid, you can either glue the grid with 1 side or 2 side. I prefer to the 1 side, as you do not need adjust the eucentric position much in TEM. But you really need skills and practice.
6) I use the Gatan Dual ot Fishion miller to ion mill the samples. Gatan pips works very faster. But use a little low voltage and current.
Hope these words help you.
Changhui
---- Original message ---- } Date: Mon, 3 Apr 2006 21:13:39 -0500 } From: chao.wang-at-materials.ox.ac.uk } Subject: [Microscopy] viaWWW: help for MgO prepration and Fe oxidation } To: clei-at-uiuc.edu } } } } } ------------------------------------------------------------ ---------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 12, 27 -- From clei-at-uiuc.edu Wed Apr 5 21:32:46 2006 12, 27 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu [128.174.5.96]) 12, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k362WkOr021892 12, 27 -- for {microscopy-at-microscopy.com} ; Wed, 5 Apr 2006 21:32:46 -0500 12, 27 -- Received: from expms6.cites.uiuc.edu (expms6.cites.uiuc.edu [128.174.5.43]) 12, 27 -- by expredir5.cites.uiuc.edu (8.13.6/8.13.6) with ESMTP id k362WdCe019013; 12, 27 -- Wed, 5 Apr 2006 21:32:44 -0500 (CDT) 12, 27 -- Received: from expms6.cites.uiuc.edu (localhost.cites.uiuc.edu [127.0.0.1]) 12, 27 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 12, 27 -- with ESMTP id BHB81957; 12, 27 -- Wed, 5 Apr 2006 21:32:33 -0500 (CDT) 12, 27 -- Received: from 128.174.5.212 12, 27 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 12, 27 -- with HTTP/1.1; 12, 27 -- Wed, 5 Apr 2006 20:32:33 -0600 12, 27 -- Date: Wed, 5 Apr 2006 20:32:33 -0600 12, 27 -- From: Changhui LEI {clei-at-uiuc.edu} 12, 27 -- Subject: Re: [Microscopy] viaWWW: help for MgO prepration 12, 27 -- and Fe oxidation 12, 27 -- To: chao.wang-at-materials.ox.ac.uk 12, 27 -- Cc: microscopy-at-microscopy.com 12, 27 -- Reply-To: clei-at-uiuc.edu 12, 27 -- X-Mailer: Webmail Mirapoint Direct 3.4.8-GR 12, 27 -- MIME-Version: 1.0 12, 27 -- Message-Id: {f27b5254.a4811668.8198300-at-expms6.cites.uiuc.edu} 12, 27 -- Content-Type: text/plain; charset=us-ascii 12, 27 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both gsosinsky-at-ucsd.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: gsosinsky-at-ucsd.edu Name: Gina Sosinsky
Organization: University of California, San Diego
Title-Subject: [Filtered] 4th International Congress on Electron Tomography
Question: *** On-line Registration and Abstract submission are now open for the 4th International Congress on Electron Tomography (4ICET) to be held Nov. 5-8, 2006 at Paradise Point Resort, San Diego, Ca. ***
Deadline for early registration September 1 Deadline for late registration October 15 Deadline for abstract submission June 15, 2006
Note: Rooms at Paradise Point are on a first come / first served basis.
Please forward this email to others.
=========== 4ICET
This congress brings together biologists, biophysicists, computer scientists, mathematicians, materials scientists and electron optical instrumentation specialists for an interdisciplinary exchange of ideas focusing on advancing methods of electron tomography (ET) in biology. ET has moved from a specialized experimental technique practiced by a few laboratories to one delivering critical information to a broad community of cell biologists, structural biologists, and neuroscientists. For students, this conference presents a unique opportunity to learn about cutting-edge advances in ET applications and methodologies.
Sessions will include:
ï Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ï Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Leiden University Medical Center, Netherlands)
ï 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); Michael Rademacher (Univ. of Vermont)
ï Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ï Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ï Emerging technologies for the multiscale: cell to tissue and molecule to cell Co-chairs: D. Hanein (Burnham Institute); C. Larabell (UCSF)
Sessions will include both invited speakers and talks selected from submitted abstracts.
For further information please check out our web site: http://www.4icet.org or contact Mark H. Ellisman, Lead Congress Organizer (mark-at-ncmir.ucsd.edu) or Grace Osborne, Congress Coordinator (gosborne-at-ncmir,ucsd.edu)
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both temanalysis-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] TEM/FIB: FEI 200 single beam FIB question
Question: Hi: We are thinking of working a group that wants to add FIB capabilities to their lab to prepare semiconductor samples. They are looking into purchasing a pre-owned FEI 200 (single beam) with a Magnum column.
I have talked to a few of my contacts and they believe this instrument would be best suited to prepare TEM samples of larger technologies at best. They believe something like a pre-owned dual beam FEI 820 or similar might be a better choice for general TEM sample preparation. I don't really know the answer, but wanted to get actual users opninions.
Ideally, I would like to receive feedback of actual users of the FEI 200 with the Magnum column and see if they can easily prepare semiconductor TEM samples with smaller technologies and: 1) Approximately how long it takes to make a sample 2) What size geometries they can easily and routinely section.
If you do not use this particular instrument, but are a frequent FIB user and have a general opinion, I would like to hear from you also.
Thanks for being a great information resource, Sandra Keller
I am currently using our Zeiss axiovert 200M to observe cell comparments (mitochondria, golgi, endosomes,...) by fluorescence. For this purpose, the main "usable" objectives available are: - 40x/0,50 LD A-Plan - 100x/1,3 Oil EC Plan Neofluar
The 40x LD is useful for our life cell imaging experiments, so I don't think we should change it. But I wonder if the 100x is really the best solution. I must say that I am a little bit disappointed by the quality of my images (I also try to improve the preparation of the samples!). When i look to the "images samples" on the CD which was included with the axiovision soft, I notice that most of the pictures of cells are taken with a 63x apochromat. I searched the site of Zeiss and found an interesting 63x apochromat with a NA of 1.4 and corrected for coverglass thickness (0,17). What is your experience with the different Zeiss objectives?
My second question concerns the Apotome feature of Axiovision. It seems to do the same job as 3D deconvolution, but how does it work? Why choose one or the other?
Thank you all in advance.
Stephane-without-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Concerning the objectives, you might want to consider buying an EC Plan-NeoFluar 40x 1,3 oil. A great objective for fluorescence (offers a clear & intense image) that I even very often prefer instead of our 100x oil.
The ApoTome is based on the grid projection theory (more details can be found on the Zeiss webpage). The difference with Deconvolution software is that this device (hardware) shows you immediately, while acquiring the image, the result. The deconvolution software will only show you the result after acquisition and running the program. If than your image does not turn out nice, you'll have to restart from the acquisition on, whereas with the ApoTome, you can immediately see the result and restart if necessary.
Hope it helps a bit!
Best,
Sven Terclavers
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: vrijdag 7 april 2006 10:58 To: sven.terclavers-at-med.kuleuven.be
Dear listers,
I am currently using our Zeiss axiovert 200M to observe cell comparments (mitochondria, golgi, endosomes,...) by fluorescence. For this purpose, the main "usable" objectives available are: - 40x/0,50 LD A-Plan - 100x/1,3 Oil EC Plan Neofluar
The 40x LD is useful for our life cell imaging experiments, so I don't think we should change it. But I wonder if the 100x is really the best solution. I must say that I am a little bit disappointed by the quality of my images (I also try to improve the preparation of the samples!). When i look to the "images samples" on the CD which was included with the axiovision soft, I notice that most of the pictures of cells are taken with a 63x apochromat. I searched the site of Zeiss and found an interesting 63x apochromat with a NA of 1.4 and corrected for coverglass thickness (0,17). What is your experience with the different Zeiss objectives?
My second question concerns the Apotome feature of Axiovision. It seems to do the same job as 3D deconvolution, but how does it work? Why choose one or the other?
Thank you all in advance.
Stephane-without-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Hello, Interested if anyone knows of a substitute for poly-L-lysine in TEM. I am currently using it to coat carbon coated formvar coated Ni grids and examine plasma membranes with immunolabelling. Fingers crossed. Martyn.
Martyn Quinn Henry Wellcome Laboratory for Cell Biology Division of Biochemistry & Molecular Biology Davidson Building Faculty of Biomedical and Life Sciences University of Glasgow G12 8QQ
==============================Original Headers============================== 1, 21 -- From mq7x-at-udcf.gla.ac.uk Fri Apr 7 05:26:35 2006 1, 21 -- Received: from hillhead.cent.gla.ac.uk (hillhead.cent.gla.ac.uk [130.209.16.101]) 1, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k37AQZLs032136 1, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 05:26:35 -0500 1, 21 -- Received: from lenzie.cent.gla.ac.uk ([130.209.16.18]) 1, 21 -- by hillhead.cent.gla.ac.uk with esmtp (Exim 4.10) 1, 21 -- id 1FRoAk-0000tA-00 1, 21 -- for microscopy-at-microscopy.com; Fri, 07 Apr 2006 11:26:34 +0100 1, 21 -- Received: from salt.ibls.gla.ac.uk ([130.209.54.27] helo=gould1) 1, 21 -- by lenzie.cent.gla.ac.uk with smtp (Exim 4.50) 1, 21 -- id 1FRoAk-00020g-BR 1, 21 -- for microscopy-at-microscopy.com; Fri, 07 Apr 2006 11:26:34 +0100 1, 21 -- Message-Id: {3.0.1.32.20060407112633.0089de10-at-udcf.gla.ac.uk} 1, 21 -- X-Sender: mq7x-at-udcf.gla.ac.uk 1, 21 -- X-Mailer: Windows Eudora Light Version 3.0.1 (32) 1, 21 -- Date: Fri, 07 Apr 2006 11:26:33 +0100 1, 21 -- To: microscopy-at-microscopy.com 1, 21 -- From: Martyn Quinn {mq7x-at-udcf.gla.ac.uk} 1, 21 -- Subject: Substitute for poly-L-lysine in TEM? 1, 21 -- Mime-Version: 1.0 1, 21 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both twigg-at-estd.nrl.navy.mil as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: twigg-at-estd.nrl.navy.mil Name: Mark Twigg
Organization: Naval Research Laboratory
Title-Subject: [Filtered] Ion Milling of SiC TEM samples
Question: I have a perennial problem in ion milling 4H-SiC TEM samples: the thin edge of the electron-transparent region often suffers from crater-like variations in thickness. These divots seem to serve as the focus for bend contours and sample bending that make diffraction contrast analysis and HRTEM imaging difficult at best. Does anybody have any ion milling prescriptions to eliminate this problem?
HI Stephane, We have a Zeiss Axiovert 100 and and Axiovert 200M in our Facility. The 100 is our confocal instrument. We have the 63x/1.4 NA oil/DIC lens on both microscopes. It performs beautifully. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 24 -- From lcgould-at-med.cornell.edu Fri Apr 7 07:46:13 2006 1, 24 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k37CkDTs020777 1, 24 -- for {microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 07:46:13 -0500 1, 24 -- Received: from mpx2.med.cornell.edu (pc113142-10.med.cornell.edu [140.251.11.119]) 1, 24 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k37CkCjv012304 1, 24 -- for {microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 08:46:12 -0400 (EDT) 1, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 24 -- by mpx2.med.cornell.edu 1, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 24 -- with ESMTPA id {0IXC007B7RGZ6Q20-at-mpx2.med.cornell.edu} for 1, 24 -- microscopy-at-microscopy.com; Fri, 07 Apr 2006 08:46:12 -0400 (EDT) 1, 24 -- Date: Fri, 07 Apr 2006 08:41:08 -0400 1, 24 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 24 -- Subject: Re: [Microscopy] Zeiss Axiovert 200M: objectives and axiovision 1, 24 -- questions 1, 24 -- In-reply-to: {200604070855.k378tLoB012849-at-ns.microscopy.com} 1, 24 -- Sender: lcgould-at-med.cornell.edu 1, 24 -- To: nizets2-at-yahoo.com, Microscopy Listserver {microscopy-at-microscopy.com} 1, 24 -- Message-id: {p06230901c05c0daee468-at-[140.251.48.23]} 1, 24 -- MIME-version: 1.0 1, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 24 -- References: {200604070855.k378tLoB012849-at-ns.microscopy.com} 1, 24 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.4.7.53106 ==============================End of - Headers==============================
If what you want is to get rid of hydrophobicity, then Alcian Blue works, float the grid for a couple of minutes on a 0.1% aqueous solution, rinse briefly and dry. Other surfactant style wetting agents are Bacitracin and BSA. Make your solution to 25 microgram/ml with Bacitracin and mount your grids. Sorry, I don't remember the concentration of BSA, but it would be about the same.
Alternatively, hydrophobicity 'fades', and if you wait several weeks the grids are essentially the same as routing formvar.
If the issue is getting the material onto the grids you can try several methods. First would be direct centrifugation to the grid. Use a Beckman Airfuge with the EM 90 Rotor. Yeah, like no one saw that reference coming, right. No, I have no commercial interest in Beckman, but was involved in some of the work demonstrating utility of the rotor. If you don't have an Airfuge available, try agar diffusion. Both methods will concentrate the membranes down onto the grid, but given the nature of the samples you are using, I suspect Agar diffusion would work better, especially coupled with Bacitracin. References are Hammond et al, 1981, J. Clin. Micro., 14:210-221, for the Airfuge, and Anderson and Doane, 1972, Appl. Microbiol., 24:495-496.
If you want, I do have the concentration protocols in electronic file form and can share them with you. Sorry, the others are not. Feel free to call if you want to discuss the procedures.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Phone: 204-789-3313 Pager: 204-931-954 Home Phone: 204-489-6924 Cell: 204-781-1502 Fax: 204-789-3926/204-489-6924
==============================Original Headers============================== 13, 20 -- From paul_hazelton-at-umanitoba.ca Fri Apr 7 09:05:48 2006 13, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k37E5lvX008389 13, 20 -- for {microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 09:05:48 -0500 13, 20 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 13, 20 -- (authenticated bits=0) 13, 20 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k37E5kgk012615; 13, 20 -- Fri, 7 Apr 2006 09:05:46 -0500 (CDT) 13, 20 -- Message-ID: {443671B6.5050109-at-umanitoba.ca} 13, 20 -- Date: Fri, 07 Apr 2006 09:05:42 -0500 13, 20 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 13, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 13, 20 -- X-Accept-Language: en-us, en 13, 20 -- MIME-Version: 1.0 13, 20 -- To: mq7x-at-udcf.gla.ac.uk 13, 20 -- Subject: Re: [Microscopy] Substitute for poly-L-lysine in TEM? 13, 20 -- References: {200604071028.k37ASWrL003471-at-ns.microscopy.com} 13, 20 -- In-Reply-To: {200604071028.k37ASWrL003471-at-ns.microscopy.com} 13, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I utilise the Alpha Plan-Fluar x100 1.45 NA objective lens on my Axiovert 200M for live cell imaging of microtubual dynamics in S.pombe, nuclear pore assembly and mitochondrial dynamics in single mammalian cells, over time periods of an hour to three days. I must admit to worrying about maintenance of the lens-oil-coverslip interface over such a time frame but the oil becoming too runny or drying up over this time frame does not seem to be a problem. Originally all of our work was carried out using the x63 apochromat but the x100 1.45 NA lens is much more light efficient.
I agree with Sven, the EC Plan-NeoFluar 40x 1.3 oil is a great lens, we utilise this for cell invasion assays and can image samples for three to five days. As well as heating the sample (Bioptechs), CO2/Air and environmental chamber (Solent), we also heat the objective lens with the Bioptechs objective heater.
We were so impressed with the alpha-plan lens we purchased another for the Olympus based Deltavision system,
All the best
Steve
Steve Bagley Associate Scientist Advanced Imaging Facility Paterson Institute for Cancer Research Cancer Research UK University of Manchester Wilmslow Road Manchester M20 4BX, UK
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: 07 April 2006 09:59 To: Steve Bagley
Dear listers,
I am currently using our Zeiss axiovert 200M to observe cell comparments (mitochondria, golgi, endosomes,...) by fluorescence. For this purpose, the main "usable" objectives available are: - 40x/0,50 LD A-Plan - 100x/1,3 Oil EC Plan Neofluar
The 40x LD is useful for our life cell imaging experiments, so I don't think we should change it. But I wonder if the 100x is really the best solution. I must say that I am a little bit disappointed by the quality of my images (I also try to improve the preparation of the samples!). When i look to the "images samples" on the CD which was included with the axiovision soft, I notice that most of the pictures of cells are taken with a 63x apochromat. I searched the site of Zeiss and found an interesting 63x apochromat with a NA of 1.4 and corrected for coverglass thickness (0,17). What is your experience with the different Zeiss objectives?
My second question concerns the Apotome feature of Axiovision. It seems to do the same job as 3D deconvolution, but how does it work? Why choose one or the other?
Thank you all in advance.
Stephane-without-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the University of Manchester. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
==============================Original Headers============================== 20, 28 -- From SBagley-at-PICR.man.ac.uk Fri Apr 7 09:55:44 2006 20, 28 -- Received: from probity.mcc.ac.uk (probity.mcc.ac.uk [130.88.200.94]) 20, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k37Eth0j018425 20, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 09:55:43 -0500 20, 28 -- Received: from jill.picr.man.ac.uk ([130.88.233.248] helo=sanmail.picr.man.ac.uk) 20, 28 -- by probity.mcc.ac.uk with esmtp (Exim 4.60 (FreeBSD)) 20, 28 -- (envelope-from {SBagley-at-PICR.man.ac.uk} ) 20, 28 -- id 1FRsND-0003su-3L 20, 28 -- for Microscopy-at-microscopy.com; Fri, 07 Apr 2006 15:55:43 +0100 20, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.3790.1830 20, 28 -- Content-class: urn:content-classes:message 20, 28 -- MIME-Version: 1.0 20, 28 -- Content-Type: text/plain; 20, 28 -- charset="us-ascii" 20, 28 -- Importance: normal 20, 28 -- Priority: normal 20, 28 -- Subject: RE: [Microscopy] Zeiss Axiovert 200M: objectives and axiovision questions 20, 28 -- Date: Fri, 7 Apr 2006 15:55:42 +0100 20, 28 -- Message-ID: {BAA35444B19AD940997ED02A6996AAE002A7BE78-at-sanmail.picr.man.ac.uk} 20, 28 -- X-MS-Has-Attach: 20, 28 -- X-MS-TNEF-Correlator: 20, 28 -- Thread-Topic: [Microscopy] Zeiss Axiovert 200M: objectives and axiovision questions 20, 28 -- Thread-Index: AcZaIaYcLhOYRwlgSCGxIooq6mB5IwAChkrA 20, 28 -- From: "Steve Bagley" {SBagley-at-PICR.man.ac.uk} 20, 28 -- To: {Microscopy-at-microscopy.com} 20, 28 -- X-UoM: Scanned by the University Mail System. See http://www.itservices.manchester.ac.uk/email/filtering/information/ for details. 20, 28 -- Content-Transfer-Encoding: 8bit 20, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k37Eth0j018425 ==============================End of - Headers==============================
Dear all, I need to demonstrate bacteria on the antibacterial cellulose substrate; the antibacterial agent is assumingly kills bacteria on the contact, and my goal is to demonstrate collapsed membrane. I have problem with the E.coli and S.aureus attachment. Any suggestions on the improving the adhesion would be highly appreciated. Thank you, Albina PS thank you all for the helpful advice on vapor fixation!
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 4, 21 -- From amich-at-ufl.edu Fri Apr 7 12:38:17 2006 4, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k37HcHcV031129 4, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 12:38:17 -0500 4, 21 -- Received: from osgjas02.cns.ufl.edu (osgjas02.cns.ufl.edu [128.227.74.132]) 4, 21 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k37HcD4i3170342 4, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 13:38:13 -0400 4, 21 -- Message-ID: {73562524.302501144431493039.JavaMail.osg-at-osgjas02.cns.ufl.edu} 4, 21 -- Date: Fri, 7 Apr 2006 13:38:13 -0400 (EDT) 4, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 4, 21 -- To: Microscopy-at-microscopy.com 4, 21 -- Subject: bacterial adhesion to the substrate 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 4, 21 -- Content-Transfer-Encoding: 7bit 4, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 4, 21 -- X-Originating-IP: 72.155.88.211 [72.155.88.211] 4, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both bliss5-at-llnl.gov as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: bliss5-at-llnl.gov Name: R. Ann Bliss
Organization: Lawrence Livermore National Lab
Title-Subject: [Filtered] Lacy grids
Question: I was about to give a colleague ordering information for lacey grids when another mentioned finding silicone contamination on these grids in the past. Has anyone found this to be true? Is there one brand better/worse than the others?
You could reply off-list if this is not an appropriate question for publication.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both jerzy.gazda-at-ceriumlabs.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: jerzy.gazda-at-ceriumlabs.com Name: Jerzy Gazda
Organization: CeriumLabs
Title-Subject: [Filtered] CCD cameras for TEM
Question: Hi, I am looking for suggestions (including from vendors - off list) for a digital camera for TEM imaging on JEM2010 used primary for metrology (2kx2K maximum, imaging speed is of essence). The camera needs to be installed below viewing chamber on the instrument and come with modern acquizition software. I am aware of three major competitors in that market, but might there be others?
R. Ann Bliss asked the following: ================================================================== Question: I was about to give a colleague ordering information for lacey grids when another mentioned finding silicone contamination on these grids in the past. Has anyone found this to be true? Is there one brand better/worse than the others?
You could reply off-list if this is not an appropriate question for publication. ================================================================== In my opinion, it is always an "appropriate" question for discussion if a quality issue could have an impact, especially if undesirable, on the quality of one's results.
It has been our experience that there are two main sources for potential Si contamination on either carbon or holey carbon or lacey carbon coated grids:
a) the vacuum evaporator itself, if it has been used for the evaporation of SiOx. It is extremely difficult to remove the last vestiges of the evaporated SiOx from previous evaporations. Another, but not so common source is the case where someone thinks they should be using silicone fluid in a DP of a vacuum evaporator with the obvious potential for contamination. It has not been uncommon for me to find that silicone grease instead of a pure hydrocarbon grease (e.g. Apiezon M or L or Santovac 5GB high vacuum grease) is sometimes being used on the bell jar o ring.
b) TEM grid storage boxes which apparently in some instances have been molded by molders who, in an effort to speed up the molding, and reduce costs, use a silicone release agent.
Another source for Si can be from the carbon rods if one is not using the very highest spectrographic purity (which do of course cost more than rods with lesser purity). As a further comment, most laboratories with vacuum evaporators have only one, and with the passing of time, it is hard to know exactly what has and has not been evaporated in them over the years. If the system is turbo pumped, there would not be any chance of silicone fluid contamination but when it comes to the carbon rods, we know that not all laboratories are using rods of the highest purity for their evaporations. I don't want to sound like I am promoting the use of turbo pumped systems since we are hard pressed to show that there is any real difference in the quality of the carbon films produced, turbo vs. DP systems.
SPI Supplies is a major manufacturer of custom coated grids for EM. All carbon evaporations are done in high vacuum systems that have never been used for SiOx evaporation and have never been used with silicone fluids or greases. And grid storage boxes sold by SPI Supplies or BEEM, to our knowledge, have never been molded with the aid of any silicone release agents. The carbon source is carbon rods of spectrographic purity. We have had, from time to time, and I mean perhaps once a year, a customer tell us that they are detecting Si on their coated grids. But we have never been able to reproduce, in our own TEM/EDS instrumentation, evidence for that contamination ourselves using retained samples from the same batch. There have been times when we have duplicated the observation of Si on coated grids that were returned to us, but not from the retained samples, causing us to conclude that the contamination occurred after the grids left SPI Supplies.
We are as baffled by this as anyone else and any additional clues as to where such Si contamination comes from would be helpful. So far as we know, Si contamination has not been a problem associated with the coated grids from SPI Supplies.
Disclaimer: SPI Supplies is a long time manufacturer of custom coated grids and further information can be found on URL http://www.2spi.com/catalog/grids/cusctgrd.html
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 15, 27 -- From cgarber-at-2spi.com Sat Apr 8 15:08:03 2006 15, 27 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 15, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k38K83kE023640 15, 27 -- for {microscopy-at-msa.microscopy.com} ; Sat, 8 Apr 2006 15:08:03 -0500 15, 27 -- Received: from yourb27fb1c401 ([71.225.86.11]) 15, 27 -- (authenticated bits=0) 15, 27 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id k38K7vWp024262; 15, 27 -- Sat, 8 Apr 2006 16:08:01 -0400 15, 27 -- X-IDV-FirstRcvd: [71.225.86.11] 15, 27 -- X-IDV-HELO: yourb27fb1c401 15, 27 -- X-IDV-Authenticated-User: cgarber 15, 27 -- Message-ID: {01d201c65b48$21a4f6f0$6501a8c0-at-yourb27fb1c401} 15, 27 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 15, 27 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 15, 27 -- Cc: {bliss5-at-llnl.gov} 15, 27 -- Subject: Silicone contamination on lacey carbon filmed grids 15, 27 -- Date: Sat, 8 Apr 2006 16:07:57 -0400 15, 27 -- MIME-Version: 1.0 15, 27 -- Content-Type: text/plain; 15, 27 -- format=flowed; 15, 27 -- charset="iso-8859-1"; 15, 27 -- reply-type=original 15, 27 -- Content-Transfer-Encoding: 7bit 15, 27 -- X-Priority: 3 15, 27 -- X-MSMail-Priority: Normal 15, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 15, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both w_d_howell-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: w_d_howell-at-yahoo.com Name: Bill Howell
Organization: Stanford
Title-Subject: [Filtered] Comparison of Zeiss objectives
Question: Hello all,
Is there any practical advantage to the Zeiss Plan-Apochromat 5X/0.16 M27 (420630-9900-000) relative to the Zeiss EC Plan-Neofluar 5X/0.15 M27 (420330-9900-000)? My intended application is general histology/pathology examining primarily H&E and immunohistochemistry slides, but also taking some low magnification digital photos. M27 refers to the screw thread on the objective.
Also, what are the relative merits of the Zeiss Plan-Apochromat 40X/0.95 Corr (dry) M27 (420660-9970-000) compared to Zeiss C-Plan-Apochromat 40X/1.20 Corr UV-VIS-IR (water) M27 (421767-9970-000)? The intended applications will be the same as above, but will definitely be used for digital photos and DIC. There is some possiblity that the objectives may be used for confocal or Apotome (structured illumination) work at some point in the future.
How was this Si contamination detected and/or characterized?
How were the grids stored. It is important to distinguish possible manufacturing issues from inappropriate handling.
Nestor Your Friendly Neighborhood SysOp
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 13 -- From zaluzec-at-microscopy.com Sun Apr 9 09:38:10 2006 7, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 7, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k39EcAaJ010045 7, 13 -- for {microscopy-at-microscopy.com} ; Sun, 9 Apr 2006 09:38:10 -0500 7, 13 -- Mime-Version: 1.0 7, 13 -- Message-Id: {p06110417c05ecb795b4e-at-[206.69.208.22]} 7, 13 -- In-Reply-To: {200604081554.k38Fs3Mn001579-at-ns.microscopy.com} 7, 13 -- References: {200604081554.k38Fs3Mn001579-at-ns.microscopy.com} 7, 13 -- Date: Sun, 9 Apr 2006 09:38:09 -0500 7, 13 -- To: microscopy-at-microscopy.com 7, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 7, 13 -- Subject: Si Contamination on Lacy grids 7, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
New York Microscopical Society 30 North Mountain Avenue Montclair, NJ 07042
Bernard Friedman Memorial Workshop
Polarized Light Microscopy April 29, May 6, 13 & 20, 2006
An advanced course on polarized light microscopy which will cover the following topics: The nature of polarized light The origin and interpretation of interference colors Birefringence and crystal orientation, The Indicatrix Compensation and variable compensators Interference figures and their interpretation
The workshop will consist of four Consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy. The course instructors include Jan Hinsch of Leica, Inc., Mary McCann of McCann Imaging, John Reffner of SensIR and N.Y.M.S. Instructor Don O'Leary.
WHEN: April 2, May 6, 13 & 20, 2006 from 10 A.M. to 4 P.M.
COST: $395 for N.Y.M.S. members, $425 for non-members (includes membership). Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants. Return form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201)797-8849 e-mail dkoleary-at-verizon.net
PLEASE POST -------------------------------------------------------------------------- Registration Form Polarized Light Microscopy
Hello, Many thanks for the suggestions and to those asking sensibly 'why?'. To clarify a bit: I am glow discharging the grids before applying the poly-L. The poly-L is being used as an adhesive. Unfortunately I may be getting some form of artefact as a result of an interaction between the negative stain and the poly-L (?). I was hoping to be able to utilize a different 'adhesive' and pinpoint the problem. Many thanks, Martyn. Martyn Quinn Henry Wellcome Laboratory for Cell Biology Division of Biochemistry & Molecular Biology Davidson Building Faculty of Biomedical and Life Sciences University of Glasgow G12 8QQ
==============================Original Headers============================== 1, 21 -- From mq7x-at-udcf.gla.ac.uk Mon Apr 10 05:42:16 2006 1, 21 -- Received: from hillend.cent.gla.ac.uk (hillend.cent.gla.ac.uk [130.209.16.102]) 1, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3AAgGJT011349 1, 21 -- for {microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 05:42:16 -0500 1, 21 -- Received: from lenzie.cent.gla.ac.uk ([130.209.16.18]) 1, 21 -- by hillend.cent.gla.ac.uk with esmtp (Exim 4.10) 1, 21 -- id 1FStqZ-0000eR-00 1, 21 -- for microscopy-at-microscopy.com; Mon, 10 Apr 2006 11:42:15 +0100 1, 21 -- Received: from db241-26.ibls.gla.ac.uk ([130.209.54.26] helo=lab204) 1, 21 -- by lenzie.cent.gla.ac.uk with smtp (Exim 4.50) 1, 21 -- id 1FStqX-0003R7-Hk 1, 21 -- for microscopy-at-microscopy.com; Mon, 10 Apr 2006 11:42:15 +0100 1, 21 -- Message-Id: {3.0.5.32.20060410114213.007f7330-at-udcf.gla.ac.uk} 1, 21 -- X-Sender: mq7x-at-udcf.gla.ac.uk 1, 21 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.5 (32) 1, 21 -- Date: Mon, 10 Apr 2006 11:42:13 +0100 1, 21 -- To: microscopy-at-microscopy.com 1, 21 -- From: Martyn Quinn {mq7x-at-udcf.gla.ac.uk} 1, 21 -- Subject: Substitute for poly-L in TEM - more info. 1, 21 -- Mime-Version: 1.0 1, 21 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Ah, what kind of staining artefact are you getting then? And what staining method do you use?
I have not yet experienced staining artefacts related to the poly-L- lysine coating in plasma membrane sheet preparation, but maybe the problem lies somewhere else in the method?
Regards,
Cornelia Muncke
EM-Unit Physiological Laboratory University of Liverpool Crown Street Liverpool L69 3BX
} } Hello, } Many thanks for the suggestions and to those asking sensibly } 'why?'. } To clarify a bit: } I am glow discharging the grids before applying the poly-L. } The poly-L is being used as an adhesive. } Unfortunately I may be getting some form of artefact as a } result of } an interaction between the negative stain and the poly-L (?). } I was hoping to be able to utilize a different 'adhesive' and } pinpoint the problem. } Many thanks, } Martyn. } Martyn Quinn } Henry Wellcome Laboratory for Cell Biology } Division of Biochemistry & Molecular Biology } Davidson Building } Faculty of Biomedical and Life Sciences } University of Glasgow } G12 8QQ }
==============================Original Headers============================== 9, 28 -- From c.muncke-at-liverpool.ac.uk Mon Apr 10 07:01:38 2006 9, 28 -- Received: from mx2.liv.ac.uk (mx2.liv.ac.uk [138.253.100.180]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3AC1cNM021883 9, 28 -- for {Microscopy-at-Microscopy.Com} ; Mon, 10 Apr 2006 07:01:38 -0500 9, 28 -- Received: from mailhuba.liv.ac.uk ([138.253.100.36]) 9, 28 -- by mx2.liv.ac.uk with esmtp (Exim 4.54) 9, 28 -- id 1FSv5J-0000ts-BO 9, 28 -- for Microscopy-at-Microscopy.Com; Mon, 10 Apr 2006 13:01:33 +0100 9, 28 -- Received: from localhost ([127.0.0.1] helo=mailhuba.liv.ac.uk) 9, 28 -- by mailhuba.liv.ac.uk with esmtp (Exim 4.54) 9, 28 -- id 1FSv5J-0000B5-AV 9, 28 -- for Microscopy-at-Microscopy.Com; Mon, 10 Apr 2006 13:01:33 +0100 9, 28 -- Received: from pc028200.med.liv.ac.uk ([138.253.28.200]) 9, 28 -- by mailhuba.liv.ac.uk with esmtpsa (TLSv1:RC4-SHA:128) 9, 28 -- (Exim 4.54) 9, 28 -- id 1FSv5J-0000Av-9t 9, 28 -- for Microscopy-at-Microscopy.Com; Mon, 10 Apr 2006 13:01:33 +0100 9, 28 -- Mime-Version: 1.0 (Apple Message framework v749.3) 9, 28 -- In-Reply-To: {200604101049.k3AAnZSG020912-at-ns.microscopy.com} 9, 28 -- References: {200604101049.k3AAnZSG020912-at-ns.microscopy.com} 9, 28 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 28 -- Message-Id: {FD2EFD1B-D948-46A2-B0E1-9FFA084BDBB5-at-liverpool.ac.uk} 9, 28 -- Content-Transfer-Encoding: 7bit 9, 28 -- From: Cornelia Muncke {c.muncke-at-liverpool.ac.uk} 9, 28 -- Subject: Re: [Microscopy] Substitute for poly-L in TEM - more info. 9, 28 -- Date: Mon, 10 Apr 2006 12:58:28 +0100 9, 28 -- To: Microscopy-at-Microscopy.Com 9, 28 -- X-Mailer: Apple Mail (2.749.3) ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both anatolebeams-at-abdm.co.uk as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: We have a Pixera PVC100C camera that we haven't used for a while. Now that we come to need it the PhotoShop plug-in has been lost in an upgrade on the Mac it was originally installed on. Pixera won't supply one as they no longer support the camera. Would anyone, by any chance, be able to supply us with just the PhotoShop plug-in, or know where we might find one. I imagine it is more-or-less the same for all their cameras.
I contribute this note only to broaden the discussion a little.
I confess to have accused my grid supplier, some years ago, of substituting SiOx coated grids for the C-coated grids I ordered. Further investigation of our lab (not UConn) uncovered a TEM user who routinely stuck her grids to the milling post with vacuum grease. This Si-based grease was then transferred to the TEM sample holder.
A thorough cleaning of sample holder and change in sample prep protocol cleared up this problem.
Cheers
-- Roger A. Ristau, PhD TEM Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269 vox: 860-486-5379 fax: 860-486-4745
==============================Original Headers============================== 7, 19 -- From raristau-at-ims.uconn.edu Mon Apr 10 09:23:36 2006 7, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3AENY9g010310 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 09:23:35 -0500 7, 19 -- Received: from [137.99.20.202] (d20h202.public.uconn.edu [137.99.20.202]) 7, 19 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k3AEN2J2026507 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 10:23:02 -0400 7, 19 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 7, 19 -- Date: Mon, 10 Apr 2006 10:22:30 -0400 7, 19 -- Subject: re: Silicone contamination on lacey carbon filmed grids 7, 19 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 7, 19 -- To: {microscopy-at-microscopy.com} 7, 19 -- Message-ID: {C05FE266.B79%raristau-at-ims.uconn.edu} 7, 19 -- Mime-version: 1.0 7, 19 -- Content-type: text/plain; charset="US-ASCII" 7, 19 -- Content-transfer-encoding: 7bit 7, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 7, 19 -- X-UConn-MailScanner: Found to be clean 7, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
The New England Society for Microscopy is pleased to announce its annual spring meeting, incorporating a workshop and the Spring Symposium, to be held at the Marine Biological Laboratory at Woods Hole, MA, May 4-6 2006.
The workshop, held on Thursday 4th. May will, this year, be on the topic of Confocal Microscopy. The symposium, held on Friday and Saturday, May 5-6, will feature, amongst other things, a panel and open discussion, moderated by John Mackenzie, on the topic of "The ethics of digital imaging and storage". The usual exhibit by the Society's corporate members will take place on Saturday morning. There will be a poster session, for which submissions are solicited.
Full details of the meeting are available on the Society's web pages, at http://nesm.cims.harvard.edu by following the link to "upcoming events".
Please note that advance registration for both events is essential. The registration deadline for the workshop is Friday April 21st. Registration for the symposium dinner must be received by Thursday April 20th. Room reservations at MBL are available on a first-come basis.
Tony Garratt-Reed - for NESM
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
The other day I mistakenly destroyed relavent information regarding a SEM digital image. I instantly realized what I had done (... what a blunder!), and I should have known better. The mistake reminded me I had intended to read the article entitled "Ethics and Digital Imaging" (Microscopy Today, V14,N1, Jan06).
The article's recommendations surprised me a bit relative to what I had considered a severe blunder. That is, I had not altered the image in any way, other than to make minor adjustments to brightness, contrast and gamma (i.e., adjustments the article would lead us to believe are relatively innocent, and do not significantly alter the image's statement of evidence). I actually agree with the committee's recommendations; so, what did I do that was so wrong? I SAVED the file with the SAME filename! In doing so, I had completely wiped pertinent information in the file format that the SEM had written to the TIFF format. Such information would have been important for duplicating the image, and under what circumstances the image was acquired (e.g., keV, mag, tilt, detector, chamber pressure, etc ... in fact, a veritable wealth of information).
Like many of you, I was aware of the information, and should have known better. It is now my practice for the SEM to write to a protected directory. Relative to user retrieval, is now a read only directory. This subject I expect the MSA sub-committee will address in a future report.
Regarding the reproducibility of image presentations, I also believe it's worth mentioning several other issues that should have been addressed:
The first issue is that softwares, like Photoshop, are capable of writing the history of modifications to the file (as well to a separate text file). Actually, I am not aware of any other software with this capability, but the possibility is an open standard for the TIFF file format. It is there for any other software to implement. It may be that the MSA sub-committee didn't want to suggest that we use a specific software, but I do believe they should have mentioned the possibility, and that it would have been that more "ethical" to use software that enabled the capability. Not to suggest this method is perfect, but it is much easier than making the same entries in your lab notes.
My last issue is a long and more complex rant, but it is connected to what makes documenting the file's history possible. It is also about the microscope specific information I mistakenly (however "innocently") obliterated. That is, I don't know of a single SEM manufacturer who takes advantage of the TIFF file format such that these accidents do not happen. The problem (IMHO) is that SEM manufacturers take advantage of the flexibility of the TIF format, but in a way that only they know how to read. Some SEMs will simply append the data onto the end of the file, and others will embed it in the header (examples included below). There is at least one 3rd party software that knows how to read at least one SEM TIFF format, but given the popularity, unique versatility, and availability of Photoshop (and other softwares), this data should be standardized and written to the TIFF such that it will remain safely. A case in point is the metadata, made popular by today's digital cameras, that most (if not all) TIFF softwares respect and will re-write to the TIFF file, including JPEGs. Furthermore, Photoshop does not need to know it is there. Photoshop will simply recognize the beginning and end of the XMP data, and re-write it when the file is saved If XMP fields are created and standardized, it will make it easy for microscopy softwares (and Photoshop compatible plug-ins) to find and use.
Lastly ... I have no connection with Adobe relative to recommending their products. I am sure I'm not alone in recognizing Adobe's contributions to digital photography, supported by a huge user-base and many professionals. Adobe also comes up with good ideas and provides a open forum and a means for creating standards when no one else will. For concluding my rant in a small way, can I suggest we expand this MSA subcommittee's mandate to look into, ask this microscope community for suggestions for the types of data, and follow the likes of NASA who created standards for embedding GPS information in their image files.
Inco Innovation Centre c/o Memorial University 230 Elizabeth Avenue P.O. Box 4200 St. John's, NL A1C 5S7
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ FEI data (appended to end of file) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ [User] User=XTLIB Usertext=Quanta W UsertextUnicode=5100750061006E007400610020005700 Date=03/31/2004 Time=01:29PM
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Zeiss data (embedded in header) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 1 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 2 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 3 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 36 AP_APERTURESIZE Aperture Size = 100.0 µm AP_BEAM_CURRENT Beam Current = 100.0 µA DP_RUNUPSTATE Beam State = Beam On AP_BEAM_TIME Beam Time = 106.72 Hours AP_BRIGHTNESS Brightness = 48.3 % AP_CHAMBER_PRESSURE Chamber = 1.30e-003 Pa AP_COLLECTOR_BIAS Collector Bias = 250 V AP_CONTRAST Contrast = 26.6 % AP_FRAME_TIME Cycle Time = 40.3 Secs AP_DATE Date :11 Mar 2004 AP_ACTUALKV EHT = 20.00 kV DP_FIXED_APERTURE2 EP Aperture = None DP_EP_MODE EP Mode = Dry AP_ACTUALCURRENT Fil I = 2.660 A AP_FILAMENT_AGE Filament Age = 18.42 Hours DP_FILAMENT_TYPE Filament Type = W (Agar A054) DP_FIXED_APERTURE Fixed Aperture (VP) = Yes AP_FRAME_AVERAGE_COUNT Frames to average = 1 AP_FRAME_INT_COUNT Frames to Int. = 0 AP_IPROBE I Probe = 208 pA AP_LINE_AVERAGE_COUNT Line Avg.Count = 4 AP_LINE_INT_COUNT Line int. count = 0 AP_MAG Mag = 84 X DP_OUT_DEV Output dev = 19/21 inch display DP_OUT_TYPE Output To = Display/File AP_HCSTAGE_TEMP Peltier Temp = 20.0 °C DP_PIXEL_SIZE Pix Size state = calibrated AP_PIXEL_SIZE Pixel Size = 4.157 µm DP_DETECTOR_CHANNEL Signal A = SE1 DP_IMPLIED_DETECTOR Signal B = SE1 AP_STAGE_AT_X Stage at X = 54.327 mm AP_STAGE_AT_Y Stage at Y = 40.286 mm AP_STAGE_AT_Z Stage at Z = 15.481 mm DP_USER User = Busy SV_USER_NAME User Name = CAMKR SV_USER_TEXT User Text = text
==============================Original Headers============================== 24, 20 -- From Michael-at-Shaffer.net Mon Apr 10 11:22:12 2006 24, 20 -- Received: from ws6-3.us4.outblaze.com (ws6-3.us4.outblaze.com [205.158.62.199]) 24, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3AGM9d5031317 24, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 11:22:10 -0500 24, 20 -- Received: (qmail 20070 invoked from network); 10 Apr 2006 16:22:08 -0000 24, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 24, 20 -- by ws6-3.us4.outblaze.com with SMTP; 10 Apr 2006 16:22:06 -0000 24, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 24, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 24, 20 -- Subject: Ethics & Digital Imaging (long) 24, 20 -- Date: Mon, 10 Apr 2006 13:52:04 -0230 24, 20 -- Message-ID: {002001c65cba$e8d76320$b995fea9-at-roamingwolf} 24, 20 -- MIME-Version: 1.0 24, 20 -- Content-Type: text/plain; 24, 20 -- charset="iso-8859-1" 24, 20 -- X-Mailer: Microsoft Office Outlook 11 24, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, 20 -- Thread-Index: AcZcuuc5vNdzeoaRTj+XFLjeiFV89g== 24, 20 -- Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3AGM9d5031317 ==============================End of - Headers==============================
Your points are very well taken. It turns out that ImageJ has a "macro recorder" that will track operations that are carried out on images and, just like Photoshop, allow you to convert that into a programmable routine. It is also possible to save this record as a text file, which is, however, independent of the original image file. This, of course, applies specifically to the post-processing component of image generation.
} } } } ---------------------------------------------------------------------- } ------ 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 other day I mistakenly destroyed relavent information regarding a } SEM digital image. I instantly realized what I had done (... what a } blunder!), and I should have known better. The mistake reminded me I } had intended to read the article entitled "Ethics and Digital Imaging" } (Microscopy Today, V14,N1, Jan06). } } The article's recommendations surprised me a bit relative to what I } had considered a severe blunder. That is, I had not altered the } image in any way, other than to make minor adjustments to brightness, } contrast and gamma (i.e., adjustments the article would lead us to } believe are relatively innocent, and do not significantly alter the } image's statement of evidence). I actually agree with the committee's } recommendations; so, what did I do that was so wrong? I SAVED the } file with the SAME filename! In doing so, I had completely wiped } pertinent information in the file format that the SEM had written to } the TIFF format. Such information would have been important for } duplicating the image, and under what circumstances the image was } acquired (e.g., keV, mag, tilt, detector, chamber pressure, etc ... in } fact, a veritable wealth of information). } } Like many of you, I was aware of the information, and should have } known better. It is now my practice for the SEM to write to a } protected directory. Relative to user retrieval, is now a read only } directory. This subject I expect the MSA sub-committee will address } in a future report. } } Regarding the reproducibility of image presentations, I also believe } it's worth mentioning several other issues that should have been } addressed: } } The first issue is that softwares, like Photoshop, are capable of } writing the history of modifications to the file (as well to a } separate text file). Actually, I am not aware of any other software } with this capability, but the possibility is an open standard for the } TIFF file format. It is there for any other software to implement. } It may be that the MSA sub-committee didn't want to suggest that we } use a specific software, but I do believe they should have mentioned } the possibility, and that it would have been that more "ethical" to } use software that enabled the capability. Not to suggest this method } is perfect, but it is much easier than making the same entries in your } lab notes. } } My last issue is a long and more complex rant, but it is connected to } what makes documenting the file's history possible. It is also about } the microscope specific information I mistakenly (however } "innocently") obliterated. That is, I don't know of a single SEM } manufacturer who takes advantage of the TIFF file format such that } these accidents do not happen. The problem (IMHO) is that SEM } manufacturers take advantage of the flexibility of the TIF format, but } in a way that only they know how to read. Some SEMs will simply append } the data onto the end of the file, and others will embed it in the } header (examples included below). There is at least one 3rd party } software that knows how to read at least one SEM TIFF format, but } given the popularity, unique versatility, and availability of } Photoshop (and other softwares), this data should be standardized and } written to the TIFF such that it will remain safely. A case in point } is the metadata, made popular by today's digital cameras, that most } (if not all) TIFF softwares respect and will re-write to the TIFF } file, including JPEGs. Furthermore, Photoshop does not need to know } it is there. Photoshop will simply recognize the beginning and end of } the XMP data, and re-write it when the file is saved If XMP fields } are created and standardized, it will make it easy for microscopy } softwares (and Photoshop compatible plug-ins) to find and use. } } Lastly ... I have no connection with Adobe relative to recommending } their products. I am sure I'm not alone in recognizing Adobe's } contributions to digital photography, supported by a huge user-base } and many professionals. Adobe also comes up with good ideas and } provides a open forum and a means for creating standards when no one } else will. For concluding my rant in a small way, can I suggest we } expand this MSA subcommittee's mandate to look into, ask this } microscope community for suggestions for the types of data, and follow } the likes of NASA who created standards for embedding GPS information } in their image files. } } genuinely :o) } michael shaffer } } SEM/MLA Research Coordinator } (709) 737-6799 (ofc) } (709) 737-6790 (lab) } (709) 737-6193 (FAX) } {http://www.mun.ca/creait/maf/} } {http://www.esd.mun.ca/epma/} } } Inco Innovation Centre } c/o Memorial University } 230 Elizabeth Avenue } P.O. Box 4200 } St. John's, NL A1C 5S7 } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } FEI data (appended to end of file) } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } [User] } User=XTLIB } Usertext=Quanta W } UsertextUnicode=5100750061006E007400610020005700 } Date=03/31/2004 } Time=01:29PM } } [SYSTEM] } DNumber=D7500 } Software=2.2 } Source=W-Tetrode } Column=W-ESEM } FinalLens=W-ESEM } Chamber= } Stage= } Pump=TMP } ESEM= } Aperture=Fixed } Scan=dispb1.0 } Acq=ViperQuad1.0 } EucWD=0.01 } } [Beam] } HV=20000 } Spot=6 } StigmatorX=0.39061 } StigmatorY=0.446543 } BeamShiftX=0 } BeamShiftY=0 } ScanRotation=0 } ImageMode=HR } } [Scan] } InternalScan=true } Dwelltime=0.0001 } FrameTime=94.251 } PixelHeight=2.14827e-006 } PixelWidth=2.14827e-006 } Horfieldsize=0.00219983 } Verfieldsize=0.00189907 } Average=0 } Integrate=0 } } [Stage] } StageX=0.00296175 } StageY=0.00145804 } StageZ=0.0100002 } StageR=3.14107 } StageT=-0.000855211 } Spectilt= } WorkingDistance=0.00999534 } } [Vacuum] } UserMode=Highvacuum } CHPressure=90 } Gas= } } [Specimen] } Temperature= } } [Detectors] } Number=1 } Name=SSD } } [SSD] } Contrast=72.9 } Brightness=39 } Signal=BSE } Mix=100 } State=true } Active=true } Mode=0 } ContrastDB=72.9 } BrightnessDB=39 } SegmentMode=0 } Setting=A+B } ContrastSpotsizeAlignment=1 } MinimumDetectorDwellTime=1e-006 } [PrivateFei] } BitShift=0 } Databarheight=0 } DataBarSelected=HV Spot WD Sig Mag HFW MicronBar Label } TimeOfCreation=31.03.2004 13:41:17 } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Zeiss data (embedded in header) } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 4.157207e-006 } 8.409719e+001 } 6 } 2.000000e+004 } 2.660000e+000 } 2.080923e-010 } 8.303549e-003 } 1 } 4.157207e-006 } 8.409719e+001 } 6 } 2.000000e+004 } 2.660000e+000 } 2.080923e-010 } 8.303549e-003 } 2 } 4.157207e-006 } 8.409719e+001 } 6 } 2.000000e+004 } 2.660000e+000 } 2.080923e-010 } 8.303549e-003 } 3 } 4.157207e-006 } 8.409719e+001 } 6 } 2.000000e+004 } 2.660000e+000 } 2.080923e-010 } 8.303549e-003 } 36 } AP_APERTURESIZE } Aperture Size = 100.0 µm } AP_BEAM_CURRENT } Beam Current = 100.0 µA } DP_RUNUPSTATE } Beam State = Beam On } AP_BEAM_TIME } Beam Time = 106.72 Hours } AP_BRIGHTNESS } Brightness = 48.3 % } AP_CHAMBER_PRESSURE } Chamber = 1.30e-003 Pa } AP_COLLECTOR_BIAS } Collector Bias = 250 V } AP_CONTRAST } Contrast = 26.6 % } AP_FRAME_TIME } Cycle Time = 40.3 Secs } AP_DATE } Date :11 Mar 2004 } AP_ACTUALKV } EHT = 20.00 kV } DP_FIXED_APERTURE2 } EP Aperture = None } DP_EP_MODE } EP Mode = Dry } AP_ACTUALCURRENT } Fil I = 2.660 A } AP_FILAMENT_AGE } Filament Age = 18.42 Hours } DP_FILAMENT_TYPE } Filament Type = W (Agar A054) } DP_FIXED_APERTURE } Fixed Aperture (VP) = Yes } AP_FRAME_AVERAGE_COUNT } Frames to average = 1 } AP_FRAME_INT_COUNT } Frames to Int. = 0 } AP_IPROBE } I Probe = 208 pA } AP_LINE_AVERAGE_COUNT } Line Avg.Count = 4 } AP_LINE_INT_COUNT } Line int. count = 0 } AP_MAG } Mag = 84 X } DP_OUT_DEV } Output dev = 19/21 inch display } DP_OUT_TYPE } Output To = Display/File } AP_HCSTAGE_TEMP } Peltier Temp = 20.0 °C } DP_PIXEL_SIZE } Pix Size state = calibrated } AP_PIXEL_SIZE } Pixel Size = 4.157 µm } DP_DETECTOR_CHANNEL } Signal A = SE1 } DP_IMPLIED_DETECTOR } Signal B = SE1 } AP_STAGE_AT_X } Stage at X = 54.327 mm } AP_STAGE_AT_Y } Stage at Y = 40.286 mm } AP_STAGE_AT_Z } Stage at Z = 15.481 mm } DP_USER } User = Busy } SV_USER_NAME } User Name = CAMKR } SV_USER_TEXT } User Text = text } } } } ==============================Original } Headers============================== 24, 20 -- From } Michael-at-Shaffer.net Mon Apr 10 11:22:12 2006 24, 20 -- Received: from } ws6-3.us4.outblaze.com (ws6-3.us4.outblaze.com [205.158.62.199]) 24, } 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k3AGM9d5031317 24, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr } 2006 11:22:10 -0500 24, 20 -- Received: (qmail 20070 invoked from } network); 10 Apr 2006 16:22:08 -0000 24, 20 -- Received: from unknown } (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 24, 20 -- } by ws6-3.us4.outblaze.com with SMTP; 10 Apr 2006 16:22:06 -0000 24, 20 } -- From: "michael shaffer" {michael-at-Shaffer.net} 24, 20 -- To: "MSA } Microscopy list" {Microscopy-at-microscopy.com} 24, 20 -- Subject: Ethics } & Digital Imaging (long) 24, 20 -- Date: Mon, 10 Apr 2006 13:52:04 } -0230 24, 20 -- Message-ID: } {002001c65cba$e8d76320$b995fea9-at-roamingwolf} 24, 20 -- MIME-Version: } 1.0 24, 20 -- Content-Type: text/plain; 24, 20 -- } charset="iso-8859-1" 24, 20 -- X-Mailer: Microsoft Office Outlook 11 } 24, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, } 20 -- Thread-Index: AcZcuuc5vNdzeoaRTj+XFLjeiFV89g== 24, 20 -- } Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id k3AGM9d5031317 } ==============================End of - } Headers==============================
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
==============================Original Headers============================== 8, 28 -- From jbs-at-temple.edu Mon Apr 10 13:20:13 2006 8, 28 -- Received: from po-smtp4.temple.edu (po-smtp4.temple.edu [155.247.166.232]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3AIKAUi010611 8, 28 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 13:20:12 -0500 8, 28 -- Received: from [155.247.98.40] (jbs.bio.temple.edu [155.247.98.40]) 8, 28 -- by po-smtp4.temple.edu (MOS 3.7.1-GA) 8, 28 -- with ESMTP id CPX28192 (AUTH jbs); 8, 28 -- Mon, 10 Apr 2006 14:19:58 -0400 (EDT) 8, 28 -- From: "Joel Sheffield" {jbs-at-temple.edu} 8, 28 -- To: michael-at-Shaffer.net, Microscopy-at-microscopy.com 8, 28 -- Date: Mon, 10 Apr 2006 14:22:17 -0400 8, 28 -- MIME-Version: 1.0 8, 28 -- Subject: Re: [Microscopy] Ethics & Digital Imaging (long) 8, 28 -- Reply-to: jbs-at-temple.edu 8, 28 -- Message-ID: {443A6A19.30006.6C074275-at-jbs.temple.edu} 8, 28 -- Priority: normal 8, 28 -- In-reply-to: {200604101622.k3AGMMHs031528-at-ns.microscopy.com} 8, 28 -- X-mailer: Pegasus Mail for Windows (4.30 public beta 1) 8, 28 -- Content-type: text/plain; charset=ISO-8859-1 8, 28 -- Content-description: Mail message body 8, 28 -- X-Junkmail-Status: score=10/50, host=po-smtp4.temple.edu 8, 28 -- X-Junkmail-SD-Raw: score=unknown, 8, 28 -- refid=str=0001.0A090207.443AA0AF.006F,ss=1,fgs=0, 8, 28 -- ip=155.247.98.40, 8, 28 -- so=2005-09-30 22:39:37, 8, 28 -- dmn=5.1.5/2006-02-08 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k3AIKAUi010611 ==============================End of - Headers==============================
} Michael, } } Your points are very well taken. It turns out that ImageJ } has a "macro recorder" that will track operations that are } carried out on images and, just like Photoshop, allow you to } convert that into a programmable routine. It is also } possible to save this record as a text file, which is, } however, independent of the original image file.
Yes ... There many possibilities, and I use ImageJ as well. With respect to a macro however, what if you made a call to an automatic binary thresholding routine? I dare say it would not save the threshold value that it had automatically determined. Photoshop document "history" however will. But, like I said, this implimentation isn't perfect either because it won't report the history IF the operations were done via an PS action (...ARGH!... And I had such high hopes!). It will however report the name of the action used.
michael shaffer :o)
} ---------------------------------------------------------------------- } } ------ The Microscopy ListServer -- CoSponsor: The } Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver On-Line Help
==============================Original Headers============================== 7, 21 -- From Michael-at-Shaffer.net Mon Apr 10 13:50:19 2006 7, 21 -- Received: from ws6-3.us4.outblaze.com (ws6-3.us4.outblaze.com [205.158.62.199]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3AIoIrM021257 7, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 13:50:19 -0500 7, 21 -- Received: (qmail 30715 invoked from network); 10 Apr 2006 18:50:18 -0000 7, 21 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 7, 21 -- by ws6-3.us4.outblaze.com with SMTP; 10 Apr 2006 18:50:18 -0000 7, 21 -- From: "michael shaffer" {michael-at-Shaffer.net} 7, 21 -- To: {jbs-at-temple.edu} , {Microscopy-at-microscopy.com} 7, 21 -- Subject: RE: [Microscopy] Ethics & Digital Imaging (long) 7, 21 -- Date: Mon, 10 Apr 2006 16:20:15 -0230 7, 21 -- Message-ID: {002f01c65ccf$9c811a10$b995fea9-at-roamingwolf} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="iso-8859-1" 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 7, 21 -- In-Reply-To: {443A6A19.30006.6C074275-at-jbs.temple.edu} 7, 21 -- Thread-Index: AcZcy2RyuKJ3mMPNSIaeE1mjdAbW2AAA0a1g 7, 21 -- Content-Transfer-Encoding: 8bit 7, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3AIoIrM021257 ==============================End of - Headers==============================
My big problem/peeve, with TIFF files and microscope formats are not limited to SEM issues.
I don't buy the proprietary sales pitch anymore. Fixing the code to embed the data in the header is trivial and could be done by a competent programmer in an afternoon. Do we as a collective group have enough sway with the industry to get them to agree and write the files with collection information included? One problem for the Hitachi 2700 we've got would be it is a passive scan system and only records what ever data was on the screen. A simple prompt much like AMT's form field might go along way to getting the data into the header file that might other wise be lost/forgotten. A counter point would certainly be that Hitachi, JEOL, FEI for example use different terminology for the same parameters. Which would become standard, if we went to a standard (i.e.: probe current, beam current, spot size).
Like I said initially, the format is minor compared to the variety of bit depth issues. Sure you can use Metamorph, or ImagePro, or ImageJ to look at a 12 bit image, but if you take the same file to Photoshop it's a hit or miss if the way the Bit Depth is recorded is compatible. What I mean is that some like Olympus write the 12 bit images along the 16 bit scale and the software knows it is an Olympus file and displays it properly, but photoshop *can* open it, and once rescaled in 16 bit mode it can be viewed easily. Leica (and Zeiss I believe) chose to create a '12-bit' TIFF file that is actually a 16 bit file. Photoshop freaks out when it tries to open it. And by freaks out I mean it displays this: "Could not compete your request because the TIFF file uses an unsupported bit depth." If you delve into the header in Metamorph (which has the programming to make sense of the crazy file formats) it finds the "Bits per Sample = 16" and the following line defining "Used Bits per Sample = 12." Yes there are reasons and some suggest that these are the 'best' ways around dealing with a system running with 12 bit hardware and the constraints of a 16 bit system. Maybe I'm too much of a perfectionist to take that as the answer. But the subject cuts to the core of Ethics and Digital Imaging (IMHO - or IMnsHO).
With that said, I am on a personal crusade against 8 bit, in an age where memory and drive space is at an all time low cost and transfer rates and processing speeds are at an incredible high (go try and copy a 1 meg file off a floppy drive), why are files still being saved at a sub-optimal bit depth? If the system collects data at a 12 or 16 bit depth, why cut the arms and legs off the image and truncate it to 8? The quip of "it is easier to use the 8 bit data" is most irksome. Users with that retort frustrate me, because the systems we have in the facility, for the most part, have software that the users can load in limited capacity on their own systems and review and edit their images. But framed in light of the previous paragraph, there is a bit of a balancing act in the middle favoring 8-bit that supports the *user's quips.*
The header/system info is a minor issues compared to bit-depth and image information. At least with film the choice of levels was acceptable even at the lowest/cheapest level.
One the positive side, I have noticed an increasing move towards more consistent TIFF file formats. It isn't great but it is better than 'back in the day' when Photoshop 4.0 was hot off the floppy disks.
(thank you for the opportunity to vent on a current frustration) Oh and I'll go ahead and plug my NESM colleague's symposium at Woods Hole on "The Ethics of Digital Imaging and Storage." I for one am eager to hear what the panelists will be saying!
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: michael-at-Shaffer.net [mailto:michael-at-Shaffer.net] Sent: Monday, April 10, 2006 12:26 PM To: Williams, Geoffrey
The other day I mistakenly destroyed relavent information regarding a SEM digital image. I instantly realized what I had done (... what a blunder!), and I should have known better. The mistake reminded me I had intended to read the article entitled "Ethics and Digital Imaging" (Microscopy Today, V14,N1, Jan06).
The article's recommendations surprised me a bit relative to what I had considered a severe blunder. That is, I had not altered the image in any way, other than to make minor adjustments to brightness, contrast and gamma (i.e., adjustments the article would lead us to believe are relatively innocent, and do not significantly alter the image's statement of evidence). I actually agree with the committee's recommendations; so, what did I do that was so wrong? I SAVED the file with the SAME filename! In doing so, I had completely wiped pertinent information in the file format that the SEM had written to the TIFF format. Such information would have been important for duplicating the image, and under what circumstances the image was acquired (e.g., keV, mag, tilt, detector, chamber pressure, etc ... in fact, a veritable wealth of information).
Like many of you, I was aware of the information, and should have known better. It is now my practice for the SEM to write to a protected directory. Relative to user retrieval, is now a read only directory. This subject I expect the MSA sub-committee will address in a future report.
Regarding the reproducibility of image presentations, I also believe it's worth mentioning several other issues that should have been addressed:
The first issue is that softwares, like Photoshop, are capable of writing the history of modifications to the file (as well to a separate text file). Actually, I am not aware of any other software with this capability, but the possibility is an open standard for the TIFF file format. It is there for any other software to implement. It may be that the MSA sub-committee didn't want to suggest that we use a specific software, but I do believe they should have mentioned the possibility, and that it would have been that more "ethical" to use software that enabled the capability. Not to suggest this method is perfect, but it is much easier than making the same entries in your lab notes.
My last issue is a long and more complex rant, but it is connected to what makes documenting the file's history possible. It is also about the microscope specific information I mistakenly (however "innocently") obliterated. That is, I don't know of a single SEM manufacturer who takes advantage of the TIFF file format such that these accidents do not happen. The problem (IMHO) is that SEM manufacturers take advantage of the flexibility of the TIF format, but in a way that only they know how to read. Some SEMs will simply append the data onto the end of the file, and others will embed it in the header (examples included below). There is at least one 3rd party software that knows how to read at least one SEM TIFF format, but given the popularity, unique versatility, and availability of Photoshop (and other softwares), this data should be standardized and written to the TIFF such that it will remain safely. A case in point is the metadata, made popular by today's digital cameras, that most (if not all) TIFF softwares respect and will re-write to the TIFF file, including JPEGs. Furthermore, Photoshop does not need to know it is there. Photoshop will simply recognize the beginning and end of the XMP data, and re-write it when the file is saved If XMP fields are created and standardized, it will make it easy for microscopy softwares (and Photoshop compatible plug-ins) to find and use.
Lastly ... I have no connection with Adobe relative to recommending their products. I am sure I'm not alone in recognizing Adobe's contributions to digital photography, supported by a huge user-base and many professionals. Adobe also comes up with good ideas and provides a open forum and a means for creating standards when no one else will. For concluding my rant in a small way, can I suggest we expand this MSA subcommittee's mandate to look into, ask this microscope community for suggestions for the types of data, and follow the likes of NASA who created standards for embedding GPS information in their image files.
Inco Innovation Centre c/o Memorial University 230 Elizabeth Avenue P.O. Box 4200 St. John's, NL A1C 5S7
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ FEI data (appended to end of file) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ [User] User=XTLIB Usertext=Quanta W UsertextUnicode=5100750061006E007400610020005700 Date=03/31/2004 Time=01:29PM
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Zeiss data (embedded in header) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 1 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 2 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 3 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 36 AP_APERTURESIZE Aperture Size = 100.0 µm AP_BEAM_CURRENT Beam Current = 100.0 µA DP_RUNUPSTATE Beam State = Beam On AP_BEAM_TIME Beam Time = 106.72 Hours AP_BRIGHTNESS Brightness = 48.3 % AP_CHAMBER_PRESSURE Chamber = 1.30e-003 Pa AP_COLLECTOR_BIAS Collector Bias = 250 V AP_CONTRAST Contrast = 26.6 % AP_FRAME_TIME Cycle Time = 40.3 Secs AP_DATE Date :11 Mar 2004 AP_ACTUALKV EHT = 20.00 kV DP_FIXED_APERTURE2 EP Aperture = None DP_EP_MODE EP Mode = Dry AP_ACTUALCURRENT Fil I = 2.660 A AP_FILAMENT_AGE Filament Age = 18.42 Hours DP_FILAMENT_TYPE Filament Type = W (Agar A054) DP_FIXED_APERTURE Fixed Aperture (VP) = Yes AP_FRAME_AVERAGE_COUNT Frames to average = 1 AP_FRAME_INT_COUNT Frames to Int. = 0 AP_IPROBE I Probe = 208 pA AP_LINE_AVERAGE_COUNT Line Avg.Count = 4 AP_LINE_INT_COUNT Line int. count = 0 AP_MAG Mag = 84 X DP_OUT_DEV Output dev = 19/21 inch display DP_OUT_TYPE Output To = Display/File AP_HCSTAGE_TEMP Peltier Temp = 20.0 °C DP_PIXEL_SIZE Pix Size state = calibrated AP_PIXEL_SIZE Pixel Size = 4.157 µm DP_DETECTOR_CHANNEL Signal A = SE1 DP_IMPLIED_DETECTOR Signal B = SE1 AP_STAGE_AT_X Stage at X = 54.327 mm AP_STAGE_AT_Y Stage at Y = 40.286 mm AP_STAGE_AT_Z Stage at Z = 15.481 mm DP_USER User = Busy SV_USER_NAME User Name = CAMKR SV_USER_TEXT User Text = text
==============================Original Headers============================== 24, 20 -- From Michael-at-Shaffer.net Mon Apr 10 11:22:12 2006 24, 20 -- Received: from ws6-3.us4.outblaze.com (ws6-3.us4.outblaze.com [205.158.62.199]) 24, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3AGM9d5031317 24, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 11:22:10 -0500 24, 20 -- Received: (qmail 20070 invoked from network); 10 Apr 2006 16:22:08 -0000 24, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 24, 20 -- by ws6-3.us4.outblaze.com with SMTP; 10 Apr 2006 16:22:06 -0000 24, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 24, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 24, 20 -- Subject: Ethics & Digital Imaging (long) 24, 20 -- Date: Mon, 10 Apr 2006 13:52:04 -0230 24, 20 -- Message-ID: {002001c65cba$e8d76320$b995fea9-at-roamingwolf} 24, 20 -- MIME-Version: 1.0 24, 20 -- Content-Type: text/plain; 24, 20 -- charset="iso-8859-1" 24, 20 -- X-Mailer: Microsoft Office Outlook 11 24, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, 20 -- Thread-Index: AcZcuuc5vNdzeoaRTj+XFLjeiFV89g== 24, 20 -- Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3AGM9d5031317 ==============================End of - Headers==============================
==============================Original Headers============================== 38, 29 -- From Geoffrey_Williams-at-brown.edu Mon Apr 10 15:03:25 2006 38, 29 -- Received: from pyxis.services.brown.edu (pyxis.services.brown.edu [128.148.106.154]) 38, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3AK3Ppd000699 38, 29 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 15:03:25 -0500 38, 29 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 38, 29 -- by pyxis.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k3AK3LoB023648 38, 29 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 16:03:21 -0400 (EDT) 38, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.31]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 38, 29 -- Mon, 10 Apr 2006 16:03:21 -0400 38, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 38, 29 -- Content-class: urn:content-classes:message 38, 29 -- MIME-Version: 1.0 38, 29 -- Content-Type: text/plain; 38, 29 -- charset="iso-8859-1" 38, 29 -- Subject: RE: [Microscopy] Ethics & Digital Imaging (long) (mine reply is a bit long too) 38, 29 -- Date: Mon, 10 Apr 2006 16:03:20 -0400 38, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F0433378E-at-MAIL1.AD.Brown.Edu} 38, 29 -- X-MS-Has-Attach: 38, 29 -- X-MS-TNEF-Correlator: 38, 29 -- Thread-Topic: [Microscopy] Ethics & Digital Imaging (long) (mine reply is a bit long too) 38, 29 -- Thread-Index: AcZcu4NHTPkBWxjxSlipd8DyO4umMgAGN24w 38, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 38, 29 -- To: {Microscopy-at-microscopy.com} 38, 29 -- X-OriginalArrivalTime: 10 Apr 2006 20:03:21.0678 (UTC) FILETIME=[D15C3EE0:01C65CD9] 38, 29 -- X-Brown-Proofpoint: Not Infected 38, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06040600 definitions=3.0.0-06041008 38, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06040600 definitions=3.0.0-06041008 38, 29 -- Content-Transfer-Encoding: 8bit 38, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3AK3Ppd000699 ==============================End of - Headers==============================
Does anyone have a favorite method by which the same cell in a culture could be imaged by fluorescence microscopy and SEM? I'm scanning the catalogs and databases for etched reference cover slips, etc., but maybe someone has "the magic wand"?
Grateful as usual for any help!
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 9, 23 -- From TindallR-at-missouri.edu Mon Apr 10 15:31:51 2006 9, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3AKVoVX011025 9, 23 -- for {microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 15:31:51 -0500 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Mon, 10 Apr 2006 15:31:49 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="us-ascii" 9, 23 -- Subject: Fluorescence/SEM correlative microscopy 9, 23 -- Date: Mon, 10 Apr 2006 15:31:48 -0500 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849ECE-at-UM-EMAIL09.um.umsystem.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Fluorescence/SEM correlative microscopy 9, 23 -- thread-index: AcZc3cqML7w+B/piQpSnZR9KLyIx9w== 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 10 Apr 2006 20:31:49.0238 (UTC) FILETIME=[CB253960:01C65CDD] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3AKVoVX011025 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both pedromfjcosta-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pedromfjcosta-at-gmail.com Name: Pedro MFJ Costa
Question: We have several tripod polishers in your lab and, unfortunately, we find that with time the precision micrometers tend to get locked up. In fact, despite trying to avoid excessive water infiltrating the micrometers they still become rusty.
I was wondering if anyone had a way to prevent this from happening or possibly to unlock them without incurring into further damage.
San Pablo - CEU University in collaboration with other five universities (Málaga, Politécnica de Madrid, País Vasco, Rey Juan Carlos, and Castilla La Mancha), nine companies, CSIC and IEEE organizes a summerschool on "Advanced Data Analysis and Modeling" in Madrid between June 26th and July 27th. The full summerschool is 120 hours long and is divided into 10 courses. Attendees may register in each course independently. The deadline for registration is June 1st. For more information, please, visit http://biocomp.cnb.csic.es/~coss/Docencia/ADAM/ADAM.htm
Best regards, Carlos Oscar
*List of courses and brief description* (full description at http://biocomp.cnb.csic.es/~coss/Docencia/ADAM/ADAM.htm)
Course 1. STATISTICAL INFERENCE (June 26th - June 29th) Introduction, Some basic statistical tests, Simple linear regression. Practical sessions: SPSS Course 2. MULTIVARIATE DATA ANALYSIS (June 26th - June 29th) Introduction, Data examination, Factor analysis, MANOVA, Multidimensional scaling, Structural equation modeling. Practical sessions: SPSS Course 3. BAYESIAN NETWORKS (July 3rd - July 6th) Basics, Inference in Bayesian networks, Learning Bayesian networks from data. Practical sessions: Hugin, Elvira, Weka, LibB Course 4. NEURAL NETWORKS (July 3rd - July 6th) Introduction, Perceptron networks, The Hebb rule, Multivariate optimization, Rule of Widrow-Hoff, Backpropagation. Practical sessions: MATLAB Course 5. ASSOCIATION RULES (July 10th - July 13th) Introduction, Rule discovering, Knowledge discoverage in biological data, Applications. Practical sessions: Bioinformatics tools Course 6. EXPERT SYSTEMS (July 10th - July 13th) Introduction, Expert system programming, Hybrid systems. Practical sessions: CLIPS and JESS Course 7. HIDDEN MARKOV MODELS (July 17th - July 20th) Introduction, Discrete HMM, Basic algorithms, Semicontinuous HMMs, Continuous HMMs, Clustering, Generalized HMMs. Practical sessions: HTK Course 8. TIME SERIES ANALYSIS (July 17th - July 20th) Introduction. Probability models, Regression and Fourier analysis, Forecasting and Data mining. Practical sessions: MATLAB, R. Course 9. DATA MINING (July 24th - July 27th) Introduction, Exploring data, Classification, Cluster analysis, Survival analysis, Anomaly detection. Practical sessions: R, WEKA Course 10. PATTERN RECOGNITION (July 24th - July 27th) Introduction, Performance of supervised classification, Preprocessing, k-nearest neighbor, classification trees, logistic regression, rule induction, combining classifiers, unsupervised classification. Practical sessions: WEKA
-- ----------------------------------------------------------- Carlos Óscar Sánchez Sorzano coss.eps-at-ceu.es Escuela Politécnica Superior Tel:+34 91 372 4034 Univ. San Pablo - CEU Fax:+34 91 372 4049 Campus Urb. Montepríncipe s/n 28668 Boadilla del Monte - Madrid http://www.uspceu.com Spain -----------------------------------------------------------
______________________________________________ LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y móviles desde 1 céntimo por minuto. http://es.voice.yahoo.com
==============================Original Headers============================== 9, 17 -- From coss.eps-at-ceu.es Tue Apr 11 02:19:58 2006 9, 17 -- Received: from smtp106.plus.mail.mud.yahoo.com (smtp106.plus.mail.mud.yahoo.com [68.142.206.239]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3B7Jwq5007236 9, 17 -- for {Microscopy-at-Microscopy.Com} ; Tue, 11 Apr 2006 02:19:58 -0500 9, 17 -- Received: (qmail 21169 invoked from network); 11 Apr 2006 07:19:57 -0000 9, 17 -- Received: from unknown (HELO ?127.0.0.1?) (cos?sorzano-at-62.14.112.193 with plain) 9, 17 -- by smtp106.plus.mail.mud.yahoo.com with SMTP; 11 Apr 2006 07:19:57 -0000 9, 17 -- Message-ID: {443B595C.7070503-at-ceu.es} 9, 17 -- Date: Tue, 11 Apr 2006 09:23:08 +0200 9, 17 -- From: =?ISO-8859-1?Q?Carlos_Oscar_S=E1nchez_Sorzano?= {coss.eps-at-ceu.es} 9, 17 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 9, 17 -- X-Accept-Language: en-us, en 9, 17 -- MIME-Version: 1.0 9, 17 -- To: Microscopy-at-Microscopy.Com 9, 17 -- Subject: Summerschool on Advanced data analysis and modelling 9, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 17 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
World famous for throwing in a grenade (just ask my friends) it never ceases to fascinate me of how popular the processing of SEM images seems to be. Go to a conference and it is only the person demonstrating Photoshop who manages to fill the lecture theatre!
Have we forgotten how to use the SEM, use the features correctly and optimise the photographic/imaging contrast and brightness? In the days when some of my clients produced 15,000 Polaroid pictures per year 90% of then looked pretty good as they came off the machine, it seems that today 50% have to be "improved" by artificial means?
Are we substituting talented computer operators for talented SEM operators, it does not seem that we are scientists in microscopy any more?
Think on :)
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 7, 29 -- From protrain-at-emcourses.com Tue Apr 11 05:30:00 2006 7, 29 -- Received: from smtp01.x-mailer.co.uk (smtp01.x-mailer.co.uk [195.13.65.37]) 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BAU0VE019274 7, 29 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 05:30:00 -0500 7, 29 -- Received: from [62.69.65.169] (helo=smtp-f.mail.legend.co.uk) 7, 29 -- by smtp01.x-mailer.co.uk with smtp (Exim 4.60) 7, 29 -- (envelope-from {protrain-at-emcourses.com} ) 7, 29 -- id 1FTG8E-0006LI-VV 7, 29 -- for microscopy-at-microscopy.com; Tue, 11 Apr 2006 11:29:59 +0100 7, 29 -- Received: (qmail 12648 invoked from network); 11 Apr 2006 10:29:48 -0000 7, 29 -- Received: from unknown (HELO advent) (212.248.148.141) 7, 29 -- by 0 with SMTP; 11 Apr 2006 10:29:48 -0000 7, 29 -- Message-ID: {00a801c65d53$50db83f0$3493f8d4-at-advent} 7, 29 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 7, 29 -- From: "Steve Chapman" {protrain-at-emcourses.com} 7, 29 -- To: "American Soc" {microscopy-at-microscopy.com} 7, 29 -- Subject: Digital Imaging in the SEM 7, 29 -- Date: Tue, 11 Apr 2006 10:42:37 +0100 7, 29 -- Organization: Protrain 7, 29 -- MIME-Version: 1.0 7, 29 -- Content-Type: text/plain; 7, 29 -- format=flowed; 7, 29 -- charset="iso-8859-1"; 7, 29 -- reply-type=original 7, 29 -- Content-Transfer-Encoding: 7bit 7, 29 -- X-Priority: 3 7, 29 -- X-MSMail-Priority: Normal 7, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2527 7, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2527 ==============================End of - Headers==============================
"Have we forgotten how to use the SEM, use the features correctly and optimize the photographic/imaging contrast and brightness? In the days when some of my clients produced 15,000 Polaroid pictures per year 90% of then looked pretty good as they came off the machine, it seems that today 50% have to be "improved" by artificial means?
Are we substituting talented computer operators for talented SEM operators,
it does not seem that we are scientists in microscopy any more?"
Let me help you pull the pin........
When I studied photomicrography my instructor, John Delly, use to tell me that a photomicrograph needs to be more than correctly exposed and illuminated. More than simply demonstrating the feature or reason for the exposure, each photograph it needs to be composed. It needs to be artistic so the viewer wants to view the image. He admitted that not every photograph will be a little artistic masterpiece, but an effort needs to be made in every exposure. I never felt there should be a difference in this approach with either light or electron images.
So the question is, if I compose, correctly expose, balance contrast and illumination levels, do I Photoshop just to Photoshop? My personal answer is that (assume correct documentation) if Photoshop, color washes, false color, unsharpmask makes a feature which already exist easier to see or understand, why not? My "client" need to understand what I see and perceive. Did we not use different developers to produce different prints, paste arrows on photos, hand tint, dot map with colored filters to enhance the information and its understanding?
Few of us have the ability to ring slides, make our own polarizing filters or weld filaments to posts or mix refractive index liquids, yet these were skills once needed. We have moved on, so it will be with imaging.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
protrain-at-emcourse s.com To: frank.karl-at-degussa.com cc: 04/11/2006 06:32 Subject: [Microscopy] Digital Imaging in the SEM AM Please respond to protrain
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hi All
World famous for throwing in a grenade (just ask my friends) it never ceases to fascinate me of how popular the processing of SEM images seems to be. Go to a conference and it is only the person demonstrating Photoshop who manages to fill the lecture theatre!
Think on :)
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 7, 29 -- From protrain-at-emcourses.com Tue Apr 11 05:30:00 2006 7, 29 -- Received: from smtp01.x-mailer.co.uk (smtp01.x-mailer.co.uk [195.13.65.37]) 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BAU0VE019274 7, 29 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 05:30:00 -0500 7, 29 -- Received: from [62.69.65.169] (helo=smtp-f.mail.legend.co.uk) 7, 29 -- by smtp01.x-mailer.co.uk with smtp (Exim 4.60) 7, 29 -- (envelope-from {protrain-at-emcourses.com} ) 7, 29 -- id 1FTG8E-0006LI-VV 7, 29 -- for microscopy-at-microscopy.com; Tue, 11 Apr 2006 11:29:59 +0100 7, 29 -- Received: (qmail 12648 invoked from network); 11 Apr 2006 10:29:48 -0000 7, 29 -- Received: from unknown (HELO advent) (212.248.148.141) 7, 29 -- by 0 with SMTP; 11 Apr 2006 10:29:48 -0000 7, 29 -- Message-ID: {00a801c65d53$50db83f0$3493f8d4-at-advent} 7, 29 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 7, 29 -- From: "Steve Chapman" {protrain-at-emcourses.com} 7, 29 -- To: "American Soc" {microscopy-at-microscopy.com} 7, 29 -- Subject: Digital Imaging in the SEM 7, 29 -- Date: Tue, 11 Apr 2006 10:42:37 +0100 7, 29 -- Organization: Protrain 7, 29 -- MIME-Version: 1.0 7, 29 -- Content-Type: text/plain; 7, 29 -- format=flowed; 7, 29 -- charset="iso-8859-1"; 7, 29 -- reply-type=original 7, 29 -- Content-Transfer-Encoding: 7bit 7, 29 -- X-Priority: 3 7, 29 -- X-MSMail-Priority: Normal 7, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2527 7, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2527 ==============================End of - Headers==============================
==============================Original Headers============================== 34, 18 -- From frank.karl-at-degussa.com Tue Apr 11 07:09:45 2006 34, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 34, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BC9hww030494 34, 18 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 07:09:44 -0500 34, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 34, 18 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k3BC9agD001475; 34, 18 -- Tue, 11 Apr 2006 14:09:37 +0200 34, 18 -- In-Reply-To: {200604111032.k3BAWZIW021656-at-ns.microscopy.com} 34, 18 -- Subject: Re: [Microscopy] Digital Imaging in the SEM 34, 18 -- To: protrain-at-emcourses.com, microscopy-at-msa.microscopy.com 34, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 34, 18 -- Message-ID: {OF5D29A84A.80CBEE9B-ON8525714D.0040ADD7-8525714D.0042C5AE-at-degussa.com} 34, 18 -- From: frank.karl-at-degussa.com 34, 18 -- Date: Tue, 11 Apr 2006 08:09:19 -0400 34, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 34, 18 -- 04/11/2006 07:09:38 AM 34, 18 -- MIME-Version: 1.0 34, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
In agreement with what Frank says, a photoshop guru will tell you, 'Photoshop can make a good image better, but it can't make a bad image good.' I find this to be true and I think we still need to instruct in the art of exposure and composition. Ultimately, using Photoshop to adjust levels is the same as using filters and dodging and burning in the old days (5 years ago). What I don't understand is where all the free time that was once spent in the darkroom has gone...
John.
frank.karl-at-degussa.com wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
--
********************************* C. John Runions, Ph.D. School of Biological and Molecular Sciences Oxford Brookes University Oxford, UK OX3 0BP
As someone who has spent a substantial portion of my life since age 12 in photographic darkrooms, as well as studying and admiring the works and methods of the "masters" of photography, it has been my perception that few prints make it out of the darkroom without being manipulated. This applies, in my experience, to artistic as well as technical images. There were a few master photographers who practiced "straight" photography, whatever that means (there are NO unmanipulated images), but wasn't it Ansel himself who compared a negative to a musical score to be interpreted by the conductor-printer? At least I think it was old Ansel. The Westons dodged and burned and manipulated freely, and it certainly wasn't for lack of techical expertise in operating their cameras and processing their films.
In my new, little has changed except the tools. We now use pixels and electrons, where we once danced around the darkroom doing elaborate jigs to cut a little light here and burn a highlight there, drawing complex diagrams of how many seconds this area gets, what contrast filter (gasp!!!! contrast filters!!!) that area gets, etc., etc., etc. In the process, we went through uncounted sheets of expensive, silver-laden paper and flushed untold gallons of nasty chemicals down the drain.
I miss all that sometimes, because there was something really peaceful and fulfilling about producing a fine silver print, especially with the soothing tones of Metallica playing in the background. But, I feel I have better control now, with less waste and time, with often superior results. If I ever get back into the darkroom, it will be as a hobby.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 9, 23 -- From TindallR-at-missouri.edu Tue Apr 11 08:57:12 2006 9, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BDvB7P019836 9, 23 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 08:57:12 -0500 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Tue, 11 Apr 2006 08:57:10 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="us-ascii" 9, 23 -- Subject: Digital Imaging in the SEM 9, 23 -- Date: Tue, 11 Apr 2006 08:57:09 -0500 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849ED4-at-UM-EMAIL09.um.umsystem.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Digital Imaging in the SEM 9, 23 -- thread-index: AcZdb9NJLUmpp7FfTHuo9NxbGK2YVQ== 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 11 Apr 2006 13:57:10.0871 (UTC) FILETIME=[D426E670:01C65D6F] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3BDvB7P019836 ==============================End of - Headers==============================
I morphed the subject to encompass all PMT collection systems, as that is really where my issue with 8 bit comes from. And it is not so much a problem with 8 bit.
That I think should be clarified.
It isn't so much a problem with the number of levels, but where the levels fall on the histogram and what the users 'attempt' to do with the levels once they collect the image.
Steve's grenade is a much appreciated catalyst. It isn't just SEM images that are the subject of over processing with Photoshop, Confocal images fit the same problem. And the systems compound the issue by pseudo coloring a black and white signal. At least in the SEM we can more intuitively relate to a black and white image (but why people would ever want to false color them for scientific purposes beyond combining various backscatter signals with a secondary signal is beyond me). Confocal users take the same PMT collection system, often with significantly fewer 'true' levels than 8-bit. So why the problem with 8 bit? Its much more challenging to get good distriminatory signal from a poor 8-bit image than a 12 or 16-bit image. 12 and 16 are exceedingly excessive bit-depths for nearly all output devices, especially for powerpoint display, so why use them?
Because students, researchers, and users don't have time, patience or the resources to learn how to do as John Delly instructed Frank to collect micrographs.
It is NOT challenging to open the awareness of the image. And it is the fundamental focus of my instruction on the SEM once we get past how to turn the beam on and align the system and focus. It absolutely is the Art in Science. And that is where 8-bit digital (not your ADC in the SEM) fails the general user population.
The faster way to correct the problem of poor images, and I don't need to tell this list that the good images are under-represented in the literature now, is increase digital bit depth storage and handling of all but the final version of the image. Coupled with a user who does not under- or over-saturate any part of the image, that will go a very long way to starting the microscopy and scientific community on the road back to aesthetically acceptable images.
The best impression "phrase" I use is to tell the people I am training, that their images reflect directly on them, their PI, and the university. If you strive to create beautiful images that have support their research the response from seminar attendees or poster session visitors will be much more positive than with a poor image. High quality, aesthetically pleasing, images will advance your career faster than poor images.
I feel we may move on (as Karl concludes) but, as long as the human eye is used for communication the demand for quality images will remain. It is only 'our' collective failing of our 'clients' that helps to accelerate the challenges in imaging.
The critical element of the discussion should not be that the darkroom was just as manipulative, because it wasn't. Darkrooms have one significant critical difference. You still (Polaroid excepted and that isn't darkroom) have an original negative to compare the information presented in the print. Also Randy's inclusion of music to the analogy begs to bring in MP3 sampling. At what sampling interval can you hear the digitization? That is a personal matter. Some people just want a bazillion songs, some want absolute clarity on the songs they have, then the speaker question little tiny ones or great big ones. All these variables make it easier a bit for many people to think in terms of photographic quality and what not.
(did I just detonate the grenade or am I just writing on borrowed time?)
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: Geoffrey_Williams-at-brown.edu [mailto:Geoffrey_Williams-at-brown.edu] Sent: Tuesday, April 11, 2006 10:21 AM To: Tindall, Randy D.
I morphed the subject to encompass all PMT collection systems, as that is really where my issue with 8 bit comes from. And it is not so much a problem with 8 bit.
That I think should be clarified.
It isn't so much a problem with the number of levels, but where the levels fall on the histogram and what the users 'attempt' to do with the levels once they collect the image.
Steve's grenade is a much appreciated catalyst. It isn't just SEM images that are the subject of over processing with Photoshop, Confocal images fit the same problem. And the systems compound the issue by pseudo coloring a black and white signal. At least in the SEM we can more intuitively relate to a black and white image (but why people would ever want to false color them for scientific purposes beyond combining various backscatter signals with a secondary signal is beyond me). Confocal users take the same PMT collection system, often with significantly fewer 'true' levels than 8-bit. So why the problem with 8 bit? Its much more challenging to get good distriminatory signal from a poor 8-bit image than a 12 or 16-bit image. 12 and 16 are exceedingly excessive bit-depths for nearly all output devices, especially for powerpoint display, so why use them?
Because students, researchers, and users don't have time, patience or the resources to learn how to do as John Delly instructed Frank to collect micrographs.
It is NOT challenging to open the awareness of the image. And it is the fundamental focus of my instruction on the SEM once we get past how to turn the beam on and align the system and focus. It absolutely is the Art in Science. And that is where 8-bit digital (not your ADC in the SEM) fails the general user population.
The faster way to correct the problem of poor images, and I don't need to tell this list that the good images are under-represented in the literature now, is increase digital bit depth storage and handling of all but the final version of the image. Coupled with a user who does not under- or over-saturate any part of the image, that will go a very long way to starting the microscopy and scientific community on the road back to aesthetically acceptable images.
The best impression "phrase" I use is to tell the people I am training, that their images reflect directly on them, their PI, and the university. If you strive to create beautiful images that have support their research the response from seminar attendees or poster session visitors will be much more positive than with a poor image. High quality, aesthetically pleasing, images will advance your career faster than poor images.
I feel we may move on (as Karl concludes) but, as long as the human eye is used for communication the demand for quality images will remain. It is only 'our' collective failing of our 'clients' that helps to accelerate the challenges in imaging.
The critical element of the discussion should not be that the darkroom was just as manipulative, because it wasn't. Darkrooms have one significant critical difference. You still (Polaroid excepted and that isn't darkroom) have an original negative to compare the information presented in the print. Also Randy's inclusion of music to the analogy begs to bring in MP3 sampling. At what sampling interval can you hear the digitization? That is a personal matter. Some people just want a bazillion songs, some want absolute clarity on the songs they have, then the speaker question little tiny ones or great big ones. All these variables make it easier a bit for many people to think in terms of photographic quality and what not.
(did I just detonate the grenade or am I just writing on borrowed time?)
Geoff Williams Leduc Bioimaging Facility Manager Brown University
Surely the simple act of selecting a contrast grade of printing paper is "manipulating" the image????
Tony
At 10:02 AM 4/11/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
There are other organizations that deal with this issue. Most of them (at least as far as I know) come to the same conclusions: The original image must be stored somewhere and must not be altered. What an original image is, however, is open to discussion. For example, a camera on a microscope may do some sharpening inside the camera. Sometimes that is not known to the users. On an SEM, you have options to influence the signal before it is shown on the screen or written to Hard disk (mix SE and BSE, for example). The consensus seems to be, that you need to store the image in that form that you first acquired and that you selected to make any sort of analysis of. In the case of an SEM this should be the image as you see it on the screen, for a camera likewise. Any processing that you do afterwards needs to be reported, but the exact details depend on who you are doing this for. I believe the forensics community has very strict guidelines regarding gamma and/or brightness, other communities may not.
How you actually achieve the above goal is probably not something a committee can make any suggestions about. Whether you store it in a read-only directory, make safety backups, or write it to CD-R directly depends on the technology you use, and any committee should not define which technology you use.
The same probably applies to making suggestions about writing the processing into a file or to the TIF header. I agree that it would be useful to have a standard way to do this, but there is no common language that can be used, nor are there even common names for the processes. One person calls a shading correction what another person calls unsharp masking. Some people use Basic as a language, others use C or Java. By defining a preference of one over the other, you would probably lose some of the capabilities that are available in different implementations. I don't think that that can be the goal of a committee. Incidentally, all our software products DO have the capability to record a history of all processing done to the images. You can even re-run the processing or apply it to another image. This is NOT a functionality that is limited to Photoshop. In addition, our software has many, many "import filters" for different SEMs, and we can probably read most of the formats out there and interpret the data correctly. If you put the images into a database that comes with the software, you can setup fields for the different pieces of information and get some safety that way. But rest assured, there is always a way someone can destroy data.
The TIF standard is very flexible, which may be the source of some of the problems, and can be confusing. For example, there is a public TIFF tag for "Resolution". However, if you use that and then try to print the image in, say, Microsoft Word, it will be interpreted literally. I.e., Word will try to print the image at the resolution that is stored there. In other words, you will see a tiny dot on the paper. That means, that applications have to find another way of storing that information, and for that they have to use "private tags" which can only be interpreted if the coding is known. I think, that a standardization of more TIF tags might help, but this then begs the next question: Image files get bigger and bigger (for example, we now acquire whole slide images, which can be GB in size). Those images cannot be dealt with effectively through TIF and other formats are needed. If a committee starts codifying the information and technology, it might stop or slow down progress in the field. Any committee has to be very careful with that. This also applies to selecting certain technologies. I have no doubt that Photoshop is a powerful product, but can it, for example, deal with GB datasets in an efficient way? Can it deal with 3D, 4D, 6D datasets (X,Y, X, time, spectral information, Fluorescence, etc.)? If not, then selecting this technology over other, perhaps not as widely accepted ones, would be a mistake.
The only way to deal with this is to allow people and companies to develop technology and let the users and markets decide what happens. This happened with TIF also. Before TIF was widely accepted, there were many different image formats, many of them proprietary. TIF had much to offer, and most companies moved to using TIF as a standard. If someone comes up with a brilliant new format that is better and more secure, I am sure that users will put pressure on companies to use that format, and the companies will eventually have to comply.
Sorry for rambling...
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: michael-at-Shaffer.net [mailto:michael-at-Shaffer.net] Sent: Monday, April 10, 2006 10:25 AM To: Mike Bode
The other day I mistakenly destroyed relavent information regarding a SEM digital image. I instantly realized what I had done (... what a blunder!), and I should have known better. The mistake reminded me I had intended to read the article entitled "Ethics and Digital Imaging" (Microscopy Today, V14,N1, Jan06).
The article's recommendations surprised me a bit relative to what I had considered a severe blunder. That is, I had not altered the image in any way, other than to make minor adjustments to brightness, contrast and gamma (i.e., adjustments the article would lead us to believe are relatively innocent, and do not significantly alter the image's statement of evidence). I actually agree with the committee's recommendations; so, what did I do that was so wrong? I SAVED the file with the SAME filename! In doing so, I had completely wiped pertinent information in the file format that the SEM had written to the TIFF format. Such information would have been important for duplicating the image, and under what circumstances the image was acquired (e.g., keV, mag, tilt, detector, chamber pressure, etc ... in fact, a veritable wealth of information).
Like many of you, I was aware of the information, and should have known better. It is now my practice for the SEM to write to a protected directory. Relative to user retrieval, is now a read only directory. This subject I expect the MSA sub-committee will address in a future report.
Regarding the reproducibility of image presentations, I also believe it's worth mentioning several other issues that should have been addressed:
The first issue is that softwares, like Photoshop, are capable of writing the history of modifications to the file (as well to a separate text file). Actually, I am not aware of any other software with this capability, but the possibility is an open standard for the TIFF file format. It is there for any other software to implement. It may be that the MSA sub-committee didn't want to suggest that we use a specific software, but I do believe they should have mentioned the possibility, and that it would have been that more "ethical" to use software that enabled the capability. Not to suggest this method is perfect, but it is much easier than making the same entries in your lab notes.
My last issue is a long and more complex rant, but it is connected to what makes documenting the file's history possible. It is also about the microscope specific information I mistakenly (however "innocently") obliterated. That is, I don't know of a single SEM manufacturer who takes advantage of the TIFF file format such that these accidents do not happen. The problem (IMHO) is that SEM manufacturers take advantage of the flexibility of the TIF format, but in a way that only they know how to read. Some SEMs will simply append the data onto the end of the file, and others will embed it in the header (examples included below). There is at least one 3rd party software that knows how to read at least one SEM TIFF format, but given the popularity, unique versatility, and availability of Photoshop (and other softwares), this data should be standardized and written to the TIFF such that it will remain safely. A case in point is the metadata, made popular by today's digital cameras, that most (if not all) TIFF softwares respect and will re-write to the TIFF file, including JPEGs. Furthermore, Photoshop does not need to know it is there. Photoshop will simply recognize the beginning and end of the XMP data, and re-write it when the file is saved If XMP fields are created and standardized, it will make it easy for microscopy softwares (and Photoshop compatible plug-ins) to find and use.
Lastly ... I have no connection with Adobe relative to recommending their products. I am sure I'm not alone in recognizing Adobe's contributions to digital photography, supported by a huge user-base and many professionals. Adobe also comes up with good ideas and provides a open forum and a means for creating standards when no one else will. For concluding my rant in a small way, can I suggest we expand this MSA subcommittee's mandate to look into, ask this microscope community for suggestions for the types of data, and follow the likes of NASA who created standards for embedding GPS information in their image files.
Inco Innovation Centre c/o Memorial University 230 Elizabeth Avenue P.O. Box 4200 St. John's, NL A1C 5S7
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ FEI data (appended to end of file) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ [User] User=XTLIB Usertext=Quanta W UsertextUnicode=5100750061006E007400610020005700 Date=03/31/2004 Time=01:29PM
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Zeiss data (embedded in header) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 1 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 2 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 3 4.157207e-006 8.409719e+001 6 2.000000e+004 2.660000e+000 2.080923e-010 8.303549e-003 36 AP_APERTURESIZE Aperture Size = 100.0 µm AP_BEAM_CURRENT Beam Current = 100.0 µA DP_RUNUPSTATE Beam State = Beam On AP_BEAM_TIME Beam Time = 106.72 Hours AP_BRIGHTNESS Brightness = 48.3 % AP_CHAMBER_PRESSURE Chamber = 1.30e-003 Pa AP_COLLECTOR_BIAS Collector Bias = 250 V AP_CONTRAST Contrast = 26.6 % AP_FRAME_TIME Cycle Time = 40.3 Secs AP_DATE Date :11 Mar 2004 AP_ACTUALKV EHT = 20.00 kV DP_FIXED_APERTURE2 EP Aperture = None DP_EP_MODE EP Mode = Dry AP_ACTUALCURRENT Fil I = 2.660 A AP_FILAMENT_AGE Filament Age = 18.42 Hours DP_FILAMENT_TYPE Filament Type = W (Agar A054) DP_FIXED_APERTURE Fixed Aperture (VP) = Yes AP_FRAME_AVERAGE_COUNT Frames to average = 1 AP_FRAME_INT_COUNT Frames to Int. = 0 AP_IPROBE I Probe = 208 pA AP_LINE_AVERAGE_COUNT Line Avg.Count = 4 AP_LINE_INT_COUNT Line int. count = 0 AP_MAG Mag = 84 X DP_OUT_DEV Output dev = 19/21 inch display DP_OUT_TYPE Output To = Display/File AP_HCSTAGE_TEMP Peltier Temp = 20.0 °C DP_PIXEL_SIZE Pix Size state = calibrated AP_PIXEL_SIZE Pixel Size = 4.157 µm DP_DETECTOR_CHANNEL Signal A = SE1 DP_IMPLIED_DETECTOR Signal B = SE1 AP_STAGE_AT_X Stage at X = 54.327 mm AP_STAGE_AT_Y Stage at Y = 40.286 mm AP_STAGE_AT_Z Stage at Z = 15.481 mm DP_USER User = Busy SV_USER_NAME User Name = CAMKR SV_USER_TEXT User Text = text
==============================Original Headers============================== 24, 20 -- From Michael-at-Shaffer.net Mon Apr 10 11:22:12 2006 24, 20 -- Received: from ws6-3.us4.outblaze.com (ws6-3.us4.outblaze.com [205.158.62.199]) 24, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3AGM9d5031317 24, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 11:22:10 -0500 24, 20 -- Received: (qmail 20070 invoked from network); 10 Apr 2006 16:22:08 -0000 24, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 24, 20 -- by ws6-3.us4.outblaze.com with SMTP; 10 Apr 2006 16:22:06 -0000 24, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 24, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 24, 20 -- Subject: Ethics & Digital Imaging (long) 24, 20 -- Date: Mon, 10 Apr 2006 13:52:04 -0230 24, 20 -- Message-ID: {002001c65cba$e8d76320$b995fea9-at-roamingwolf} 24, 20 -- MIME-Version: 1.0 24, 20 -- Content-Type: text/plain; 24, 20 -- charset="iso-8859-1" 24, 20 -- X-Mailer: Microsoft Office Outlook 11 24, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, 20 -- Thread-Index: AcZcuuc5vNdzeoaRTj+XFLjeiFV89g== 24, 20 -- Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3AGM9d5031317 ==============================End of - Headers==============================
==============================Original Headers============================== 41, 23 -- From Mike.Bode-at-olympus-sis.com Tue Apr 11 11:09:43 2006 41, 23 -- Received: from mail.soft-imaging.de (mail.soft-imaging.de [62.180.61.131]) 41, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BG9gD6027584 41, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 11:09:43 -0500 41, 23 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 41, 23 -- by mail.soft-imaging.de (8.11.6/8.11.6/SuSE Linux 0.5) with ESMTP id k3BG9g423642 41, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 18:09:42 +0200 41, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 41, 23 -- Content-class: urn:content-classes:message 41, 23 -- MIME-Version: 1.0 41, 23 -- Content-Type: text/plain; 41, 23 -- charset="iso-8859-1" 41, 23 -- Subject: Re: [Microscopy] Ethics & Digital Imaging (long) 41, 23 -- Date: Tue, 11 Apr 2006 18:06:00 +0200 41, 23 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC94174CC8-at-ms-s-gws.soft-imaging.net} 41, 23 -- X-MS-Has-Attach: 41, 23 -- X-MS-TNEF-Correlator: 41, 23 -- Thread-Topic: Re: [Microscopy] Ethics & Digital Imaging (long) 41, 23 -- Thread-Index: AcZcu11KZEmu0TY5QBm2H0F4QYS2agAA3NuwAAHWLjAALsZUoA== 41, 23 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 41, 23 -- To: {Microscopy-at-microscopy.com} 41, 23 -- Content-Transfer-Encoding: 8bit 41, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3BG9gD6027584 ==============================End of - Headers==============================
In my case, I get to blame it on the lousy monitor that came with my SEM. It is the least expensive LCD that HP makes, and its gamma ranges from 2.0 to 2.5 depending on the angle of view ... and the dark shades are worse straight-on!!! ... In any case ... A few tonal tweaks with Photoshop make the images much better, and PS also interfaces much better with printers than otherwise.
Inco Innovation Centre c/o Memorial University 230 Elizabeth Avenue P.O. Box 4200 St. John's, NL A1C 5S7
} -----Original Message----- } From: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com] } Sent: April 11, 2006 8:01 AM } To: michael-at-shaffer.net } Subject: [Microscopy] Digital Imaging in the SEM } } } } } -------------------------------------------------------------- } -------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } -------------- } } Hi All } } World famous for throwing in a grenade (just ask my friends) } it never ceases to fascinate me of how popular the processing } of SEM images seems to be. Go to a conference and it is only } the person demonstrating Photoshop who manages to fill the } lecture theatre! } } Have we forgotten how to use the SEM, use the features } correctly and optimise the photographic/imaging contrast and } brightness? In the days when some of my clients produced } 15,000 Polaroid pictures per year 90% of then looked pretty } good as they came off the machine, it seems that today 50% } have to be "improved" by artificial means? } } Are we substituting talented computer operators for talented } SEM operators, it does not seem that we are scientists in } microscopy any more? } } Think on :) } } Steve Chapman } Senior Consultant Protrain } For electron microscopy consultancy and training world wide } Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 } 606967 Web www.emcourses.com } } } ==============================Original } Headers============================== } 7, 29 -- From protrain-at-emcourses.com Tue Apr 11 05:30:00 2006 } 7, 29 -- Received: from smtp01.x-mailer.co.uk } (smtp01.x-mailer.co.uk [195.13.65.37]) } 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id k3BAU0VE019274 } 7, 29 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr } 2006 05:30:00 -0500 } 7, 29 -- Received: from [62.69.65.169] (helo=smtp-f.mail.legend.co.uk) } 7, 29 -- by smtp01.x-mailer.co.uk with smtp (Exim 4.60) } 7, 29 -- (envelope-from {protrain-at-emcourses.com} ) } 7, 29 -- id 1FTG8E-0006LI-VV } 7, 29 -- for microscopy-at-microscopy.com; Tue, 11 Apr 2006 } 11:29:59 +0100 } 7, 29 -- Received: (qmail 12648 invoked from network); 11 Apr } 2006 10:29:48 -0000 7, 29 -- Received: from unknown (HELO } advent) (212.248.148.141) } 7, 29 -- by 0 with SMTP; 11 Apr 2006 10:29:48 -0000 } 7, 29 -- Message-ID: {00a801c65d53$50db83f0$3493f8d4-at-advent} } 7, 29 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} } 7, 29 -- From: "Steve Chapman" {protrain-at-emcourses.com} 7, 29 } -- To: "American Soc" {microscopy-at-microscopy.com} 7, 29 -- } Subject: Digital Imaging in the SEM 7, 29 -- Date: Tue, 11 } Apr 2006 10:42:37 +0100 7, 29 -- Organization: Protrain 7, 29 } -- MIME-Version: 1.0 7, 29 -- Content-Type: text/plain; } 7, 29 -- format=flowed; } 7, 29 -- charset="iso-8859-1"; } 7, 29 -- reply-type=original } 7, 29 -- Content-Transfer-Encoding: 7bit 7, 29 -- X-Priority: } 3 7, 29 -- X-MSMail-Priority: Normal 7, 29 -- X-Mailer: } Microsoft Outlook Express 6.00.2900.2527 7, 29 -- X-MimeOLE: } Produced By Microsoft MimeOLE V6.00.2900.2527 } ==============================End of - } Headers============================== }
==============================Original Headers============================== 10, 20 -- From Michael-at-Shaffer.net Tue Apr 11 11:13:36 2006 10, 20 -- Received: from ws6-1.us4.outblaze.com (ws6-1.us4.outblaze.com [205.158.62.196]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3BGDZBq000655 10, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 11:13:35 -0500 10, 20 -- Received: (qmail 25930 invoked from network); 11 Apr 2006 16:13:35 -0000 10, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 10, 20 -- by ws6-1.us4.outblaze.com with SMTP; 11 Apr 2006 16:13:34 -0000 10, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 10, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 10, 20 -- Subject: RE: [Microscopy] Digital Imaging in the SEM 10, 20 -- Date: Tue, 11 Apr 2006 13:43:33 -0230 10, 20 -- Message-ID: {002901c65d82$e224ec00$37079986-at-roamingwolf} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="US-ASCII" 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- X-Mailer: Microsoft Office Outlook 11 10, 20 -- In-Reply-To: {200604111030.k3BAUs1p020086-at-ns.microscopy.com} 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 10, 20 -- Thread-Index: AcZdUwOOYC6gjj9qTYCyCv805nEsBgACIEEg ==============================End of - Headers==============================
Not FL + SEM, but FL + AFM, simultaneously. What do you need to image?
If FL+SEM, I think that there are finder stages that can be used for both. Contact Bill Miller at microbill-at-mohawk.net. He is likely to have the answer.
Good hunting.
B At 03:34 PM 4/10/2006, TindallR-at-missouri.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
... Just a curious note on this subject: Ansel Adams used a microwave for final processing of many of his images and developed quite a protocol for setting and timing... Does that constitute "manipulation"?
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
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 09:00 AM 4/11/2006, TindallR-at-missouri.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 9, 17 -- From bfoster-at-mme1.com Tue Apr 11 12:03:33 2006 9, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BH3XeF018497 9, 17 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 12:03:33 -0500 9, 17 -- Received: (qmail 9631 invoked by uid 2020); 11 Apr 2006 13:04:57 -0400 9, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 9, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 11 Apr 2006 13:04:57 -0400 9, 17 -- Message-Id: {7.0.1.0.0.20060411120143.020cd2b0-at-mme1.com} 9, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 17 -- Date: Tue, 11 Apr 2006 12:03:40 -0500 9, 17 -- To: TindallR-at-missouri.edu, Microscopy Listserver {microscopy-at-microscopy.com} 9, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 9, 17 -- Subject: Re: [Microscopy] Digital Imaging in the SEM 9, 17 -- In-Reply-To: {200604111400.k3BE0FKN024040-at-ns.microscopy.com} 9, 17 -- References: {200604111400.k3BE0FKN024040-at-ns.microscopy.com} 9, 17 -- Mime-Version: 1.0 9, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
This is not only a common problem with micrometers on our Tripod(TM) Polisher and competitor units alike, but also with some of the hand grinding and lapping units that are on the market. As you mention, the most important thing is to keep water from getting into the parts that are susceptible to rusting. Don't turn them upside down when wet and dry them after you are finished. I have owned and used Tripod(TM) polishers and both the Gatan and the Fischione hand grinding units. Because these would see use everyday, I took them apart and used a Lithium grease on the threads. If I remember correctly, the Lithium grease that I used was for outboard motors and is water repellent. (You can get this type of grease almost anywhere.) By being careful and periodically lubricating the threads and moving parts, I never had a unit fail on me from corrosion. In fact, I never had a unit fail at all. If you get a tube of this grease, with the amount that you will use, it should last about a thousand years. In other words, just use it very sparingly. If during a session, you think that you may have gotten water into the sliding portion of the micrometers or onto the threads, wait until after you are done with your session and when you are cleaning up, disassemble the units, dry them, and then reapply the thin layer of grease.
For completeness, on the South Bay Technology hand lapping fixtures, the parts that could be exposed to water are all stainless steel and the sliding surfaces have a solid film lubricant on them. With these fixtures, they can get "sticky" if water is allowed to get into the sliding surfaces. When this occurs, simply take them apart and wipe the sliding surfaces with a dry cloth or paper towel and put them together again. This will remove an oxide that forms on the solid lubricant due to the water exposure and expose good solid lubricant again. The piston should slide easily again.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: pedromfjcosta-at-gmail.com [mailto:pedromfjcosta-at-gmail.com] Sent: Monday, April 10, 2006 9:48 PM To: Walck-at-SouthBayTech.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both pedromfjcosta-at-gmail.com as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: pedromfjcosta-at-gmail.com Name: Pedro MFJ Costa
Question: We have several tripod polishers in your lab and, unfortunately, we find that with time the precision micrometers tend to get locked up. In fact, despite trying to avoid excessive water infiltrating the micrometers they still become rusty.
I was wondering if anyone had a way to prevent this from happening or possibly to unlock them without incurring into further damage.
At 12:06 PM 4/11/2006 -0500, Barbara Foster wrote: } ... Just a curious note on this subject: Ansel Adams used a microwave for } final processing of many of his images and developed quite a protocol for } setting and timing... Does that constitute "manipulation"?
I heard Ansel Adams talk about this at a conference in Asilomar years ago, but he only talked about used the microwave to quickly dry test strips prints in order to see what subtile highlight changes would be found between the wet print and the dry one.
He didn't actually manipulate with the microwave ; {) _____________________________________ Tom Gore, Advanced Imaging Laboratory Department of Biology University of Victoria Box 3020 Station CSC Victoria BC V8W 3N5 Canada voice 250 721 7134 fax 250 721 7120 web: http://web.uvic.ca/ail/
==============================Original Headers============================== 4, 20 -- From togo-at-uvic.ca Tue Apr 11 12:31:43 2006 4, 20 -- Received: from castle.comp.uvic.ca (castle.comp.uvic.ca [142.104.5.97]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BHVgp9012056 4, 20 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 12:31:43 -0500 4, 20 -- Received: from amidol (amidol.biol.uvic.ca [142.104.208.15]) 4, 20 -- by castle.comp.uvic.ca (8.13.6/8.13.6) with ESMTP id k3BHVeLN3645616 4, 20 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 10:31:40 -0700 4, 20 -- Message-Id: {4.2.0.58.20060411102951.052980f0-at-pop.uvic.ca} 4, 20 -- X-Sender: togo-at-pop.uvic.ca 4, 20 -- X-Mailer: QUALCOMM Windows Eudora Pro Version 4.2.0.58 4, 20 -- Date: Tue, 11 Apr 2006 10:38:11 -0800 4, 20 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 4, 20 -- From: Tom Gore {togo-at-uvic.ca} 4, 20 -- Subject: Ansel's microwave 4, 20 -- In-Reply-To: {200604111706.k3BH6uKL029865-at-ns.microscopy.com} 4, 20 -- Mime-Version: 1.0 4, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 4, 20 -- X-UVic-Virus-Scanned: OK - Passed virus scan by Sophos (sophie) on castle 4, 20 -- X-UVic-Spam-Scan: castle.comp.uvic.ca Not_scanned_LOCAL 4, 20 -- X-Scanned-By: MIMEDefang 2.33 (www . roaringpenguin . com / mimedefang) ==============================End of - Headers==============================
Interesting. What was the microwave used for? Heating the developer?
For any artistic images there is of course no protocol, and none would make sense, so for purely artistic images you can do whatever you want to.
For scientific images there is of course a higher standard. The basic idea of science is that experiments can be verified. That requires complete disclosure how data were obtained. For an image it means that anybody who is somewhat knowledgeable in the technique must be able to recreate all steps that lead to a certain conclusion. Since much of image processing is a destructive process (information gets thrown away and cannot be recovered), the only way to assure that is to keep a copy of the "original" image and then describe what was done to the image, so someone else can take the same image, apply the same steps and come to the same conclusion.
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: bfoster-at-mme1.com [mailto:bfoster-at-mme1.com] Sent: Tuesday, April 11, 2006 11:06 AM To: Mike Bode
... Just a curious note on this subject: Ansel Adams used a microwave for final processing of many of his images and developed quite a protocol for setting and timing... Does that constitute "manipulation"?
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
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 09:00 AM 4/11/2006, TindallR-at-missouri.edu wrote:
} ----------------------------------------------------------------------- } ----- The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
} soothing tones of Metallica playing in the background. But, I feel I } have better control now, with less waste and time, with often superior } results. If I ever get back into the darkroom, it will be as a hobby. } } Cheers, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W125 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original } Headers============================== } 9, 23 -- From TindallR-at-missouri.edu Tue Apr 11 08:57:12 2006 9, 23 -- } Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) } 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BDvB7P019836 } 9, 23 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 08:57:12 -0500 } 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 9, 23 -- Tue, 11 Apr 2006 08:57:10 -0500 } 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- } Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 } 9, 23 -- Content-Type: text/plain; } 9, 23 -- charset="us-ascii" } 9, 23 -- Subject: Digital Imaging in the SEM 9, 23 -- Date: Tue, 11 Apr
} Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: } {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 11 Apr 2006
} 13:57:10.0871 (UTC) FILETIME=[D426E670:01C65D6F] 9, 23 -- } Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id k3BDvB7P019836 } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 17 -- From bfoster-at-mme1.com Tue Apr 11 12:03:33 2006 9, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BH3XeF018497 9, 17 -- for {microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 12:03:33 -0500 9, 17 -- Received: (qmail 9631 invoked by uid 2020); 11 Apr 2006 13:04:57 -0400 9, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 9, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 11 Apr 2006 13:04:57 -0400 9, 17 -- Message-Id: {7.0.1.0.0.20060411120143.020cd2b0-at-mme1.com} 9, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 17 --
On Apr 11, 2006, at 10:33 AM, Mike.Bode-at-olympus-sis.com wrote:
} Interesting. What was the microwave used for? Heating the developer? } Dear Mike, After the prints were exposed and developed (and, I seem to remember, fixed) they were microwaved for a time to enhance the contrast. The process produced very sharp, good-looking prints. Ansel Adams was known for the amount of time, care, and use of many procedures to make what he considered the best-looking prints from his negatives. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue Apr 11 12:58:44 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BHwh7X031687 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 12:58:43 -0500 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 6606634B28 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 10:58:43 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 0B06D10AD59 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 10:54:57 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200604111733.k3BHXWwO014846-at-ns.microscopy.com} 4, 22 -- References: {200604111733.k3BHXWwO014846-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {7effee60d68b87a824bbd09d1d080e5e-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Digital Imaging in the SEM 4, 22 -- Date: Tue, 11 Apr 2006 11:04:37 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
...and beautiful pictures they are. I have a couple of reprints in my house. I didn't know that Adams was playing with microwaves...
Michael Bode
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS CORP. 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-Mail: Mike.Bode-at-olympus-sis.net www.olympus-sis.net
-----Original Message----- X-from: tivol-at-caltech.edu [mailto:tivol-at-caltech.edu] Sent: Tuesday, April 11, 2006 11:01 AM To: Mike Bode
On Apr 11, 2006, at 10:33 AM, Mike.Bode-at-olympus-sis.com wrote:
} Interesting. What was the microwave used for? Heating the developer? } Dear Mike, After the prints were exposed and developed (and, I seem to remember, fixed) they were microwaved for a time to enhance the contrast. The process produced very sharp, good-looking prints. Ansel Adams was known for the amount of time, care, and use of many procedures to make what he considered the best-looking prints from his negatives. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue Apr 11 12:58:44 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BHwh7X031687 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 12:58:43 -0500 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 6606634B28 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 10:58:43 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 0B06D10AD59 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Apr 2006 10:54:57 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200604111733.k3BHXWwO014846-at-ns.microscopy.com} 4, 22 -- References: {200604111733.k3BHXWwO014846-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {7effee60d68b87a824bbd09d1d080e5e-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Digital Imaging in the SEM 4, 22 -- Date: Tue, 11 Apr 2006 11:04:37 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
==============================Original Headers============================== 12, 24 -- From Mike.Bode-at-olympus-sis.com Tue Apr 11 13:11:44 2006 12, 24 -- Received: from mail.soft-imaging.de (mail.soft-imaging.de [62.180.61.131]) 12, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3BIBiIt009063 12, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 11 Apr 2006 13:11:44 -0500 12, 24 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 12, 24 -- by mail.soft-imaging.de (8.11.6/8.11.6/SuSE Linux 0.5) with ESMTP id k3BIBi427099; 12, 24 -- Tue, 11 Apr 2006 20:11:44 +0200 12, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 12, 24 -- Content-class: urn:content-classes:message 12, 24 -- MIME-Version: 1.0 12, 24 -- Content-Type: text/plain; 12, 24 -- charset="us-ascii" 12, 24 -- Subject: RE: [Microscopy] Re: Digital Imaging in the SEM 12, 24 -- Date: Tue, 11 Apr 2006 20:07:45 +0200 12, 24 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC94174D00-at-ms-s-gws.soft-imaging.net} 12, 24 -- X-MS-Has-Attach: 12, 24 -- X-MS-TNEF-Correlator: 12, 24 -- Thread-Topic: [Microscopy] Re: Digital Imaging in the SEM 12, 24 -- Thread-Index: AcZdkfW6jQ7Z6VoWQAyKDyjGDSEXvgAALV2A 12, 24 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 12, 24 -- To: {Microscopy-at-microscopy.com} 12, 24 -- Cc: {tivol-at-caltech.edu} 12, 24 -- Content-Transfer-Encoding: 8bit 12, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3BIBiIt009063 ==============================End of - Headers==============================
Pedro, Another possibility from the boating world is to use lanolin. Some consider it to be the best rust preventive available (used on tools kept on board). The one drawback I can see for micrometers is that the viscosity is very high and could cause some problems with the tight clearances on a good micrometer. Some experimentation would be in order. Otherwise, as mentioned, lithium grease is considered to be very waterproof and has a low viscosity.
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: pedromfjcosta-at-gmail.com [mailto:pedromfjcosta-at-gmail.com] Sent: Tuesday, April 11, 2006 12:48 AM To: kenconverse-at-qualityimages.biz
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying
please copy both pedromfjcosta-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pedromfjcosta-at-gmail.com Name: Pedro MFJ Costa
Question: We have several tripod polishers in your lab and, unfortunately, we find that with time the precision micrometers tend to get locked up. In fact, despite trying to avoid excessive water infiltrating the micrometers they still become rusty.
I was wondering if anyone had a way to prevent this from happening or possibly to unlock them without incurring into further damage.
==============================Original Headers============================== 8, 12 -- From zaluzec-at-microscopy.com Mon Apr 10 23:44:01 2006 8, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3B4i0Dm027217 8, 12 -- for {microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 23:44:00 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: (Unverified) 8, 12 -- Message-Id: {p06110403c060e47f14e0-at-[206.69.208.22]} 8, 12 -- Date: Mon, 10 Apr 2006 23:43:57 -0500 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- From: pedromfjcosta-at-gmail.com (by way of MicroscopyListserver) 8, 12 -- Subject: viaWWW: Tripod Polisher micrometers locking 8, 12 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
___________________________________________________________ $0 Web Hosting with up to 200MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
==============================Original Headers============================== 25, 24 -- From kenconverse-at-qualityimages.biz Wed Apr 12 13:11:07 2006 25, 24 -- Received: from qualityimages.biz (dpmail16.doteasy.com [65.61.209.16]) 25, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3CIB7ee022039 25, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Apr 2006 13:11:07 -0500 25, 24 -- Received: from Ken [68.64.242.101] by qualityimages.biz with ESMTP 25, 24 -- (SMTPD32-8.05) id A2BD47030146; Wed, 12 Apr 2006 11:11:09 -0700 25, 24 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 25, 24 -- To: {pedromfjcosta-at-gmail.com} , 25, 24 -- "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 25, 24 -- Subject: RE: [Microscopy] viaWWW: Tripod Polisher micrometers locking 25, 24 -- Date: Wed, 12 Apr 2006 14:11:22 -0400 25, 24 -- Message-ID: {001401c65e5c$83c4e900$6501a8c0-at-Ken} 25, 24 -- MIME-Version: 1.0 25, 24 -- Content-Type: text/plain; 25, 24 -- charset="us-ascii" 25, 24 -- X-Priority: 3 (Normal) 25, 24 -- X-MSMail-Priority: Normal 25, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 25, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 25, 24 -- Importance: Normal 25, 24 -- In-Reply-To: {200604110447.k3B4lZhZ032373-at-ns.microscopy.com} 25, 24 -- X-IMSTrailer: __IMail_7__ 25, 24 -- Content-Transfer-Encoding: 8bit 25, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3CIB7ee022039 ==============================End of - Headers==============================
} Avoid the rush and register now! } } "AFM and Other Scanned Probe Microscopies" presented at N.C. State } University in Raleigh, NC by Prof. Phil Russell, Dr. Joe Griffith, } Alexei Gruverman and others. } Lab sessions will utilize instrumentation from most major } instrumentation vendors. } } This one-week short course has evolved from the numerous Scanned Probe } Microscopy courses developed and taught by Prof. Russell over the past } 2 decades. It is designed for technicians, scientists, engineers, and } researchers. The course includes laboratories with hands-on time using } a variety of scanning probe microscope (SPM) systems. Each student } will receive a notebook of all materials covered in the lectures and } animation/simulation software covering AFM principles.
} For more information go to www.ncsu.edu/aif/afmcourse
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both nyilmaz-at-mersin.edu.tr as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
We're planning to study staph aureus biofilms by TEM on a latex material like examination gloves. Does anybody know any method for it and is ruthenium red an obligation for processing or can we do it by conventional methods? Thanks in advance...
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both robn-at-hyperbranch.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: robn-at-hyperbranch.com Name: Rob Naslund
Organization: hyperbranch medical technology
Title-Subject: [Filtered] SEM - cryo-SEM
Question: I am looking for a contract lab to do some cyro-SEM work on hydrogels. Any suggestions?
We need to contact for-profit EM service providers (sample preparation and imaging, not repair and maintenance) for a financial study related to core facilities being conducted by a professor in our management school.
Would those providing these types of services please contact me off-line.
Many thanks,
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 8, 21 -- From dsherman-at-purdue.edu Fri Apr 14 08:21:59 2006 8, 21 -- Received: from exchange.purdue.edu (1061exfe04.adpc.purdue.edu [128.210.63.227]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3EDLxsa001703 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 14 Apr 2006 08:21:59 -0500 8, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.235]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 8, 21 -- Fri, 14 Apr 2006 09:21:56 -0400 8, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 8, 21 -- Fri, 14 Apr 2006 13:21:56 +0000 8, 21 -- User-Agent: Microsoft-Entourage/11.2.3.060209 8, 21 -- Date: Fri, 14 Apr 2006 09:21:56 -0400 8, 21 -- Subject: Commercial EM service 8, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 8, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 8, 21 -- Message-ID: {C0651A34.EE99%dsherman-at-purdue.edu} 8, 21 -- Thread-Topic: Commercial EM service 8, 21 -- Thread-Index: AcZfxmbUpRgsyMu5EdqNzQARJN08Mg== 8, 21 -- Mime-version: 1.0 8, 21 -- Content-type: text/plain; 8, 21 -- charset="US-ASCII" 8, 21 -- Content-transfer-encoding: 7bit 8, 21 -- X-OriginalArrivalTime: 14 Apr 2006 13:21:56.0962 (UTC) FILETIME=[67673C20:01C65FC6] ==============================End of - Headers==============================
Think the nonlinear responses of film/paper vs. the nonlinear responses of video vs. PMT point scanning vs. linearity of CCDs vs. wide dynamic range of 12+ bits. And what you see in your darkroom or monitor is not what the printer will reproduce in halftone and variable inks on variable papers or scanned or resized and compressed and thrown up on the web.
The two biggest way I deal with this are: 1. Running tests to characterize the imaging technique. 2. Forcing researchers to do control samples and image them and postprocess them at the same settings etc. as the experimental. They should be doing this anyhow. Usually the real question isn't an objective absolute answer but variations with conditions and how this fits into the bigger picture, e.g. compared with gene expression or blots for proteins or whole animal behavior etc. The photo of the KO animal's brain is meaningless without the companion photo of the normal animal's brain. Very likely, it's not so important whether the image originated as a 640 X 480 pixels digitized from video, 35mm Tri-X pan printed on Ilford grade 4 paper or a 1315 X 1000 pixel 12 bits highly linear CCD. Can we resolve the structures under scrutiny and are there meaningful differences in the biology?
In philosophy and the arts there are enormous bodies of literature that uniformly debunk the notion of absolute meanings in photographs. Regardless of the physics and purity and sanctity of the absolute object, we are interpreting beings. ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 5, 22 -- From cammer-at-aecom.yu.edu Fri Apr 14 11:38:20 2006 5, 22 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3EGcJdJ013734 5, 22 -- for {microscopy-at-microscopy.com} ; Fri, 14 Apr 2006 11:38:20 -0500 5, 22 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 5, 22 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with SMTP id k3EGc9oo026389 5, 22 -- for {microscopy-at-microscopy.com} ; Fri, 14 Apr 2006 12:38:18 -0400 5, 22 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 5, 22 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006041412381813819 5, 22 -- for {microscopy-at-microscopy.com} ; Fri, 14 Apr 2006 12:38:18 -0400 5, 22 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.30.137]) 5, 22 -- by post.aecom.yu.edu (Postfix) with ESMTP id ADE102FD0 5, 22 -- for {microscopy-at-microscopy.com} ; Fri, 14 Apr 2006 12:38:18 -0400 (EDT) 5, 22 -- Message-Id: {5.2.1.1.2.20060414123130.02a88fa8-at-mailserver.aecom.yu.edu} 5, 22 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 5, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 5, 22 -- Date: Fri, 14 Apr 2006 12:36:18 -0400 5, 22 -- To: microscopy-at-microscopy.com 5, 22 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 5, 22 -- Subject: Re: Digital Imaging in the SEM and absolute truth in imaging 5, 22 -- Mime-Version: 1.0 5, 22 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both pengw.william-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pengw.william-at-gmail.com Name: William Peng
Organization: Tsinghua University
Title-Subject: [Filtered] Profile data export in DigitalMicrograph
Question: hello,all
I want to export multiple line profile data in Gatan DigitalMicrograph to Origin software. I can change the display type from line plot to spreadsheet in object menu. A spreadsheet is showed only have intensity value and no X value. Could I get a data file about this kind of line profile?
Thanks in advance.
William Peng pengw.william-at-gmail.com ----------------------------------------------- Department of Materials Science & Engineering, Tsinghua University China
Rob Naslund wrote: ================================================================ Question: I am looking for a contract lab to do some cyro-SEM work on hydrogels. Any suggestions? ================================================================ Structure Probe, Inc has an Oxford cryo-SEM system interfaced to a JEOL Model 840 SEM. We have had experience characterizing hydrogels by this approach. Part of our firm is an independent analytical laboratory.
Contact me off-line for a proposal.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
==============================Original Headers============================== 13, 29 -- From cgarber-at-2spi.com Sun Apr 16 17:26:50 2006 13, 29 -- Received: from s-utl01-sjpop.stsn.net (s-utl01-sjpop.stsn.net [72.254.0.201]) 13, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3GMQnKb019913 13, 29 -- for {microscopy-at-msa.microscopy.com} ; Sun, 16 Apr 2006 17:26:49 -0500 13, 29 -- Received: from s-utl01-sjpop.stsn.net ([127.0.0.1]) 13, 29 -- by s-utl01-sjpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006041615264724786 13, 29 -- ; Sun, 16 Apr 2006 15:26:47 -0700 13, 29 -- X-Spam-Status: No, hits=0.0 required=9.9 13, 29 -- tests=ALL_TRUSTED: -2.867,AWL: 0.105,BAYES_00: -1.665, 13, 29 -- DATE_IN_PAST_03_06: 0.127,SARE_RECV_ADDR: 0.027 13, 29 -- X-Spam-Level: 13, 29 -- Received: from ibm1x23g2abfyg ([10.1.185.112]) 13, 29 -- by s-utl01-sjpop.stsn.net; 13, 29 -- Sun, 16 Apr 2006 15:26:46 -0700 13, 29 -- Message-ID: {001601c661a4$d37615a0$70b9010a-at-ibm1x23g2abfyg} 13, 29 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 13, 29 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 13, 29 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 13, 29 -- Subject: Cryo-SEM on hydrogels 13, 29 -- Date: Sun, 16 Apr 2006 13:31:21 -0400 13, 29 -- MIME-Version: 1.0 13, 29 -- Content-Type: text/plain; 13, 29 -- charset="Windows-1252" 13, 29 -- X-Priority: 3 13, 29 -- X-MSMail-Priority: Normal 13, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 13, 29 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 13, 29 -- Content-Transfer-Encoding: 8bit 13, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3GMQnKb019913 ==============================End of - Headers==============================
"Selecting a CCD Camera for Light Microscopy," a live, informative, and interactive web-based seminar is being held this Friday (21-April) at 11:30 AM (New York time).
Details are below. There is no cost, but connection lines are limited so reserve yours now.
-------------------------------------------------------------- TO RESERVE YOUR CONNECTION LINE -------------------------------------------------------------- Click this URL:
Click REGISTER and complete the requested information. You will be sent a link that gives you access to Friday's meeting.
---------------------------------------- MEETING SUMMARY ---------------------------------------- Name: Selecting a CCD Camera for Light Microscopy
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both pedromfjcosta-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pedromfjcosta-at-gmail.com Name: Pedro MFJ Costa
Organization: University of Cambridge
Title-Subject: [Filtered] Grease for ion millers
Question: We have a Gatan PIPS ion miller in our lab which periodically needs maintenance of the O-rings. For sealing and lubricant purposes we usually use Fomblin Vac3 grease. However, we find that it tends to be a bit too sticky for O-rings of moving parts and forces us to be often cleaning the chamber shutter. I would be grateful for suggestions on which is the best silicon-free grease to use in the Gatan PIPS, particularly as lubricant.
Pedro MFJ Costa wrote: =========================================================== Question: We have a Gatan PIPS ion miller in our lab which periodically needs maintenance of the O-rings. For sealing and lubricant purposes we usually use Fomblin Vac3 grease. However, we find that it tends to be a bit too sticky for O-rings of moving parts and forces us to be often cleaning the chamber shutter. I would be grateful for suggestions on which is the best silicon-free grease to use in the Gatan PIPS, particularly as lubricant =========================================================== You are presently using a PFPE (perfluorinated polyether) grease. It is a two component system, PTFE particles dispersed in a perfluorinated liquid continuous phase. Although the vapor pressure of the liquid phase is quite low, and is in the range of a diffusion pump fluid, it does eventually "disappear".
Braycote Micronic 803 is similar in characteristics but the grease is filtered through a screen pack filter to remove any agglomerates of the PTFE particles larger than 1 um. It is a more homogeneous lubricant. It would seem that one could expect this Braycote product to perform more to your satisfaction for the above cited reasons. You can find out more about Braycote Micronic 803 at URL http://www.2spi.com/catalog/vac/braycote-micronic-803.shtml
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
.
==============================Original Headers============================== 15, 30 -- From cgarber-at-2spi.com Tue Apr 18 04:04:05 2006 15, 30 -- Received: from s-utl01-sjpop.stsn.net (s-utl01-sjpop.stsn.net [72.254.0.201]) 15, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3I944ja000489 15, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 18 Apr 2006 04:04:05 -0500 15, 30 -- Received: from s-utl01-sjpop.stsn.net ([127.0.0.1]) 15, 30 -- by s-utl01-sjpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006041802040430529 15, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 18 Apr 2006 02:04:04 -0700 15, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 15, 30 -- tests=ALL_TRUSTED: -2.867,AWL: 0.101,BAYES_00: -1.665, 15, 30 -- DATE_IN_PAST_03_06: 0.127,SARE_RECV_ADDR: 0.027 15, 30 -- X-Spam-Level: 15, 30 -- Received: from ibm1x23g2abfyg ([10.1.185.112]) 15, 30 -- by s-utl01-sjpop.stsn.net 15, 30 -- for microscopy-at-msa.microscopy.com; 15, 30 -- Tue, 18 Apr 2006 02:04:04 -0700 15, 30 -- Message-ID: {001101c662c7$065beee0$70b9010a-at-ibm1x23g2abfyg} 15, 30 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 15, 30 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 15, 30 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 15, 30 -- Subject: Grease for ion millers 15, 30 -- Date: Mon, 17 Apr 2006 23:09:34 -0400 15, 30 -- MIME-Version: 1.0 15, 30 -- Content-Type: text/plain; 15, 30 -- charset="Windows-1252" 15, 30 -- X-Priority: 3 15, 30 -- X-MSMail-Priority: Normal 15, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 15, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 15, 30 -- Content-Transfer-Encoding: 8bit 15, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3I944ja000489 ==============================End of - Headers==============================
The locked micrometers are probably toast. When you get new micrometers, or if you want to protect ones that aren't rusted, unscrew the barrels and rub the threads with real lanolin, not a chemical substitute. Lanolin can be purchased in most drugstores. Sheep swear by the stuff! Works for tripod polisher micrometers and the turnbuckles on my sailboat, which are exposed to salt water.
Ron Anderson
pedromfjcosta-at-gmail.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both pedromfjcosta-at-gmail.com as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: pedromfjcosta-at-gmail.com } Name: Pedro MFJ Costa } } Organization: University of Cambridge } } Title-Subject: [Filtered] Tripod Polisher micrometers locking } } Question: We have several tripod polishers in your lab and, unfortunately, we find that with time the precision micrometers tend to get locked up. In fact, despite trying to avoid excessive water infiltrating the micrometers they still become rusty. } } I was wondering if anyone had a way to prevent this from happening or possibly to unlock them without incurring into further damage. } } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 8, 12 -- From zaluzec-at-microscopy.com Mon Apr 10 23:44:01 2006 } 8, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3B4i0Dm027217 } 8, 12 -- for {microscopy-at-microscopy.com} ; Mon, 10 Apr 2006 23:44:00 -0500 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: (Unverified) } 8, 12 -- Message-Id: {p06110403c060e47f14e0-at-[206.69.208.22]} } 8, 12 -- Date: Mon, 10 Apr 2006 23:43:57 -0500 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: pedromfjcosta-at-gmail.com (by way of MicroscopyListserver) } 8, 12 -- Subject: viaWWW: Tripod Polisher micrometers locking } 8, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== } } }
Pedro, My experience is that Braycote 803 works great for static seals but, like the Fomblin, becomes sticky (grabs) overnight. Braycote 602 is a different formulation with about 2 decades lower vapor pressure with moly disulfide included as a lubricant. It seems to be much better for dynamic seals. It's also a lot more expensive, but IMHO is worth it. I believe Chuck has it or can get it.
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: cgarber-at-2spi.com [mailto:cgarber-at-2spi.com] Sent: Tuesday, April 18, 2006 5:11 AM To: kenconverse-at-qualityimages.biz
Pedro MFJ Costa wrote: =========================================================== Question: We have a Gatan PIPS ion miller in our lab which periodically needs maintenance of the O-rings. For sealing and lubricant purposes we usually use Fomblin Vac3 grease. However, we find that it tends to be a bit too sticky for O-rings of moving parts and forces us to be often cleaning the chamber shutter. I would be grateful for suggestions on which is the best silicon-free grease to use in the Gatan PIPS, particularly as lubricant =========================================================== You are presently using a PFPE (perfluorinated polyether) grease. It is a two component system, PTFE particles dispersed in a perfluorinated liquid continuous phase. Although the vapor pressure of the liquid phase is quite low, and is in the range of a diffusion pump fluid, it does eventually "disappear".
Braycote Micronic 803 is similar in characteristics but the grease is filtered through a screen pack filter to remove any agglomerates of the PTFE particles larger than 1 um. It is a more homogeneous lubricant. It would seem that one could expect this Braycote product to perform more to your satisfaction for the above cited reasons. You can find out more about Braycote Micronic 803 at URL http://www.2spi.com/catalog/vac/braycote-micronic-803.shtml
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
.
==============================Original Headers============================== 15, 30 -- From cgarber-at-2spi.com Tue Apr 18 04:04:05 2006 15, 30 -- Received: from s-utl01-sjpop.stsn.net (s-utl01-sjpop.stsn.net [72.254.0.201]) 15, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k3I944ja000489 15, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 18 Apr 2006 04:04:05 -0500 15, 30 -- Received: from s-utl01-sjpop.stsn.net ([127.0.0.1]) 15, 30 -- by s-utl01-sjpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006041802040430529 15, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 18 Apr 2006 02:04:04 -0700 15, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 15, 30 -- tests=ALL_TRUSTED: -2.867,AWL: 0.101,BAYES_00: -1.665, 15, 30 -- DATE_IN_PAST_03_06: 0.127,SARE_RECV_ADDR: 0.027 15, 30 -- X-Spam-Level: 15, 30 -- Received: from ibm1x23g2abfyg ([10.1.185.112]) 15, 30 -- by s-utl01-sjpop.stsn.net 15, 30 -- for microscopy-at-msa.microscopy.com; 15, 30 -- Tue, 18 Apr 2006 02:04:04 -0700 15, 30 -- Message-ID: {001101c662c7$065beee0$70b9010a-at-ibm1x23g2abfyg} 15, 30 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 15, 30 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 15, 30 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 15, 30 -- Subject: Grease for ion millers 15, 30 -- Date: Mon, 17 Apr 2006 23:09:34 -0400 15, 30 -- MIME-Version: 1.0 15, 30 -- Content-Type: text/plain; 15, 30 -- charset="Windows-1252" 15, 30 -- X-Priority: 3 15, 30 -- X-MSMail-Priority: Normal 15, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 15, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 15, 30 -- Content-Transfer-Encoding: 8bit 15, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3I944ja000489 ==============================End of - Headers==============================
___________________________________________________________ $0 Web Hosting with up to 200MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
==============================Original Headers============================== 31, 24 -- From kenconverse-at-qualityimages.biz Tue Apr 18 10:46:46 2006 31, 24 -- Received: from qualityimages.biz (dpmail16.doteasy.com [65.61.209.16]) 31, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3IFkjR9031673 31, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 18 Apr 2006 10:46:46 -0500 31, 24 -- Received: from Ken [68.64.242.101] by qualityimages.biz with ESMTP 31, 24 -- (SMTPD32-8.05) id A9C9280800BE; Tue, 18 Apr 2006 08:46:17 -0700 31, 24 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 31, 24 -- To: {cgarber-at-2spi.com} , {pedromfjcosta-at-gmail.com} , 31, 24 -- "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 31, 24 -- Subject: RE: [Microscopy] Grease for ion millers 31, 24 -- Date: Tue, 18 Apr 2006 11:46:11 -0400 31, 24 -- Message-ID: {001201c662ff$3d4f7d80$6401a8c0-at-Ken} 31, 24 -- MIME-Version: 1.0 31, 24 -- Content-Type: text/plain; 31, 24 -- charset="us-ascii" 31, 24 -- X-Priority: 3 (Normal) 31, 24 -- X-MSMail-Priority: Normal 31, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 31, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 31, 24 -- Importance: Normal 31, 24 -- In-Reply-To: {200604180910.k3I9Aln0005869-at-ns.microscopy.com} 31, 24 -- X-IMSTrailer: __IMail_7__ 31, 24 -- Content-Transfer-Encoding: 8bit 31, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3IFkjR9031673 ==============================End of - Headers==============================
A position is open for a Transmission Electron Microscopy Analyst in the Process Characterization Group at ATDF (www.atdf.com), a wholly-owned subsidiary of SEMATECH) in Austin, Texas.
Our group provides technical support to research and development projects for SEMATECH (www.sematech.org), ATDF, and ATDF customer funded projects in the field of semiconductor and nanomaterials development. The analyst would interface with engineers and project managers, determine analytical plans, conduct analyses, and interpret and present data. The work involves characterization of materials systems new to the semiconductor industry and there is significant opportunity to do interesting and publishable science. Our business plan allows significant opportunities for collaboration and visibility within the industry as well as the materials and microscopy academic communities.
Our toolset includes two 300 keV TEM's (one FEG, one LaB6), with SUTW-EDAX EDXS, GIF-2001 EELS, multiple HAADF-STEM and CCD cameras, and a biprism paired with a Lorentz lens. Our sample preparation toolset includes several broad-beam ion mills as well as 2 focused ion beam systems.
The optimal candidate should hold a PhD in Materials Science, Chemistry or Physics, plus have several years experience in TEM characterization of semiconductor materials. A strong understanding of wafer processing is favorable.
The successful candidate should look forward to becoming an important part of our analytical team, working to solve problems, publish and present at technical conferences, lead internal teams and act in the role as a mentor to support the development of others within our group.
Interested parties should send their CV by email to Mark.Clark-at-atdf.com
==============================Original Headers============================== 8, 41 -- From Mark.Clark-at-atdf.com Tue Apr 18 11:06:31 2006 8, 41 -- Received: from outbound2-kan-R.bigfish.com (outbound-kan.frontbridge.com [63.161.60.23]) 8, 41 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3IG6VfM009275 8, 41 -- for {Microscopy-at-Microscopy.Com} ; Tue, 18 Apr 2006 11:06:31 -0500 8, 41 -- Received: from outbound2-kan.bigfish.com (localhost.localdomain [127.0.0.1]) 8, 41 -- by outbound2-kan-R.bigfish.com (Postfix) with ESMTP id 7213044C913 8, 41 -- for {Microscopy-at-Microscopy.Com} ; Tue, 18 Apr 2006 16:06:31 +0000 (UTC) 8, 41 -- Received: from mail4-kan-R.bigfish.com (unknown [172.18.7.1]) 8, 41 -- by outbound2-kan.bigfish.com (Postfix) with ESMTP id 6AC7544C9DD 8, 41 -- for {Microscopy-at-Microscopy.Com} ; Tue, 18 Apr 2006 16:06:31 +0000 (UTC) 8, 41 -- Received: from mail4-kan.bigfish.com (localhost.localdomain [127.0.0.1]) 8, 41 -- by mail4-kan-R.bigfish.com (Postfix) with ESMTP id 5F1AF377D15 8, 41 -- for {Microscopy-at-Microscopy.Com} ; Tue, 18 Apr 2006 16:06:31 +0000 (UTC) 8, 41 -- X-BigFish: VP 8, 41 -- Received: by mail4-kan (MessageSwitch) id 1145376391329369_22560; Tue, 18 Apr 2006 16:06:31 +0000 (UCT) 8, 41 -- Received: from mitch.eng.sematech.org (GATER5.SEMATECH.ORG [192.73.53.5]) 8, 41 -- by mail4-kan.bigfish.com (Postfix) with ESMTP id 4734B376DCD 8, 41 -- for {Microscopy-at-Microscopy.Com} ; Tue, 18 Apr 2006 16:06:31 +0000 (UTC) 8, 41 -- Received: from CONVERSION-DAEMON.SEMATECH.Org by SEMATECH.Org 8, 41 -- (PMDF V6.2-X26 #31211) id {01M1EG5EO8KW00VB0A-at-SEMATECH.Org} for 8, 41 -- Microscopy-at-Microscopy.Com; Tue, 18 Apr 2006 11:06:30 -0500 (CDT) 8, 41 -- Received: from cod.logon.sematech.org (cod.logon.sematech.org [131.153.32.40]) 8, 41 -- by SEMATECH.Org (PMDF V6.2-X26 #31211) 8, 41 -- with ESMTP id {01M1EG5EC7AW00VGBP-at-SEMATECH.Org} for Microscopy-at-Microscopy.Com; 8, 41 -- Tue, 18 Apr 2006 11:06:30 -0500 (CDT) 8, 41 -- Date: Tue, 18 Apr 2006 11:06:29 -0500 8, 41 -- From: Mark.Clark-at-atdf.com 8, 41 -- Subject: TEM Analyst Position Open at SEMATECH/ATDF in Austin, Texas 8, 41 -- To: Microscopy-at-Microscopy.Com 8, 41 -- Cc: Celina.Ortiz-at-atdf.com 8, 41 -- Message-id: {59157C31AE9B264D8CB608D6026FE53E016E3425-at-cod.logon.sematech.org} 8, 41 -- MIME-version: 1.0 8, 41 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 41 -- Content-type: text/plain; charset=iso-8859-1 8, 41 -- Thread-Topic: TEM Analyst Position Open at SEMATECH/ATDF in Austin, Texas 8, 41 -- Thread-Index: AcZjAg2uUy4ImXZWSK+uFP9g/A6ReQ== 8, 41 -- Content-class: urn:content-classes:message 8, 41 -- X-MS-Has-Attach: 8, 41 -- X-MS-TNEF-Correlator: 8, 41 -- Content-Transfer-Encoding: 8bit 8, 41 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3IG6VfM009275 ==============================End of - Headers==============================
I have just inherited a Hitachi S-2700 LaB6 SEM. It came from an industrial setting and some of the instructions were missing. I think they were in a desk in the room with the microscope and when the company wanted to vacate the space ASAP the movers shoved out all the easy to move furniture, including the desk with the instructions and specimen holders, but they couldn't budge the microscope so it stayed there until I picked it up.
So, I have the instructions for a tungsten filament unit, but this one is clearly a LaB6 guy. I need a clue about how to open the gun, set the tip, and if there are any operating changes for routine use.
Anybody got anything that would help?
The specimen holder stuff is all missing too. From the W instruction book I can see that there was supposed to be a jig for adjusting specimen height and some special intermediate pieces to mate the stub with the stage. I have never had a Hitachi SEM before, so I and not familiar with their system, any hints or clues would be helpful.
Thanks
Jon
-- I'm riding in the MS 150 Crusin' to the Coast bike ride May 13 - 14, 2006 to raise money for the National Multiple Sclerosis Society. You can go to http://www.msconnection.org and follow the links to make an ePledge if you wish.
Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 10, 21 -- From jmkrupp-at-ucsc.edu Tue Apr 18 12:58:43 2006 10, 21 -- Received: from smtp-prod-mx2.ucsc.edu (smtp-prod-mx2.ucsc.edu [128.114.125.44]) 10, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3IHwhQ1020869 10, 21 -- for {microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 12:58:43 -0500 10, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 10, 21 -- by smtp-prod-mx2.ucsc.edu (8.13.6/8.13.1) with ESMTP id k3IHwXCe013575 10, 21 -- for {microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 10:58:33 -0700 (PDT) 10, 21 -- Received: from [128.114.25.185] (account jmkrupp-at-ucsc.edu [128.114.25.185] verified) 10, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 10, 21 -- with ESMTPA id 52946994 for microscopy-at-microscopy.com; Tue, 18 Apr 2006 10:58:33 -0700 10, 21 -- Mime-Version: 1.0 10, 21 -- Message-Id: {p06230903c06ad716f8c0-at-[128.114.25.185]} 10, 21 -- Date: Tue, 18 Apr 2006 10:58:32 -0700 10, 21 -- To: microscopy-at-microscopy.com 10, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 10, 21 -- Subject: Hitachi S-2700 LaB6 instructions 10, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 10, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 10, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 10, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 10, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
Secure sales for BAXS Microanalysis Products in the southeast U.S. and act as company rep to existing customer base and prospective customers. Responsibilities include: prospecting for new customers, following up sales leads provided by the company, presenting and demonstrating company products, supplying quotations and technical information, formulating sales strategies, as well as negotiating and securing sales orders. In addition, the Regional Sales Manager will maintain contact with existing customers to assure their satisfaction, develop good working relationships with OEM salespeople, collect and report market information and provide routine sales forecasts.
Territory includes NC, SC, GA, FL, TN, AL, MS, AR, and LA. At least 50% travel is required. Bachelor's degree (B.S.or B.A.) from four-year college or university; or five years experience and/or training; or equivalent combination of education and experience. Three to five years experience in scientific equipment sales is a must. This position requires excellent verbal communication and interaction skills.
Bruker AXS Inc. offers a competitive salary and comprehensive benefits package including health and dental insurance, Flexible Spending Account, company sponsored life and disability insurance, 401(k) plan with company matching components, stock option plan, and vacation and sick/personal days.
Qualified applicants should submit resume and salary requirements in confidence to:
Bruker AXS Inc. Attn: Don Becker, Sales Manager 1239 Parkway Ave. Suite 203 Ewing, NJ 08628 FAX#: 609-771-4411 E-mail: don.becker-at-bruker-axs.com
Equal Opportunity Employer
==============================Original Headers============================== 10, 23 -- From Don.Becker-at-bruker-axs.com Tue Apr 18 13:06:28 2006 10, 23 -- Received: from isa1.bruker-axs.com (host-216-153-173-238.mil.choiceone.net [216.153.173.238]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3II6QWu030668 10, 23 -- for {microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 13:06:28 -0500 10, 23 -- Received: from mail1.bruker-axs.com ([172.16.0.32]) by isa1.bruker-axs.com with Microsoft SMTPSVC(6.0.3790.211); 10, 23 -- Tue, 18 Apr 2006 13:05:47 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.6944.0 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: Employment Opportunity at Bruker AXS 10, 23 -- Date: Tue, 18 Apr 2006 13:05:46 -0500 10, 23 -- Message-ID: {235D0F9F0400B3449E8C229D9D24CC570242EDD5-at-mail1.bruker-axs.com} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Employment Opportunity at Bruker AXS 10, 23 -- Thread-Index: AcZjEreJTLN9eJJZQ16cOHsBpPFsSg== 10, 23 -- From: "Becker, Don" {Don.Becker-at-bruker-axs.com} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 18 Apr 2006 18:05:47.0345 (UTC) FILETIME=[B7F47410:01C66312] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3II6QWu030668 ==============================End of - Headers==============================
Dear All, Can someone please give me paraffin-sectioning advice? I am having problems getting the paraffin to not split when I am sectioning. The sections are supposed to be done at 8 microns. I have tried cooling the block, knife and tweezers, still the sections split sometimes down the middle other times at the sides. Today I heated the block a bit, and also tried 10 microns still the sections are splitting. Please HELP !! Thank you, Kellie
--------------------------------------------------------------------------------------------------------- This e-mail message may contain privileged and/or confidential information, and is intended to be received only by persons entitled to receive such information. If you have received this e-mail in error, please notify the sender immediately. Please delete it and all attachments from any servers, hard drives or any other media. Other use of this e-mail by you is strictly prohibited.
All e-mails and attachments sent and received are subject to monitoring, reading and archival by Monsanto. The recipient of this e-mail is solely responsible for checking for the presence of "Viruses" or other "Malware". Monsanto accepts no liability for any damage caused by any such code transmitted by or accompanying this e-mail or any attachment. ---------------------------------------------------------------------------------------------------------
==============================Original Headers============================== 5, 25 -- From kellie.l.garner-at-monsanto.com Tue Apr 18 14:32:14 2006 5, 25 -- Received: from gateway1.monsanto.com (gateway1.monsanto.com [199.89.234.134]) 5, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3IJWD7w010177 5, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 14:32:14 -0500 5, 25 -- Received: from agstlsmtp03.monsanto.com (agstlsmtp03.email.monsanto.com [10.30.65.106]) 5, 25 -- by gateway1.monsanto.com (Switch-3.1.8/Switch-3.1.7) with ESMTP id k3IJWBrH003099 5, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 14:32:11 -0500 (CDT) 5, 25 -- thread-index: AcZjH0YzOS+sUgv2TgGWKHc9li8/7A== 5, 25 -- Received: from agstlhub02.monsanto.com ([10.30.64.99]) by agstlsmtp03.monsanto.com with Microsoft SMTPSVC(6.0.3790.1830); Tue, 18 Apr 2006 14:35:39 -0500 5, 25 -- Received: by agstlhub02.monsanto.com with Internet Mail Service (5.5.2653.19) id {J1VW8DGZ} ; Tue, 18 Apr 2006 14:28:21 -0500 5, 25 -- Message-ID: {30E69DC92141D14D93FAE0EC43BF9EEE0634A6-at-NA1000EXM02.na.ds.monsanto.com} 5, 25 -- From: "GARNER, KELLIE L [AG-Contractor/1000]" {kellie.l.garner-at-monsanto.com} 5, 25 -- To: {Microscopy-at-microscopy.com} 5, 25 -- Content-Transfer-Encoding: 7bit 5, 25 -- Subject: LM paraffin section methods troubleshoot 5, 25 -- Date: Tue, 18 Apr 2006 14:32:00 -0500 5, 25 -- MIME-Version: 1.0 5, 25 -- Content-Class: urn:content-classes:message 5, 25 -- X-Mailer: Internet Mail Service (5.5.2653.19) 5, 25 -- Importance: normal 5, 25 -- Priority: normal 5, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.3790.2663 5, 25 -- Content-Type: text/plain; 5, 25 -- charset="iso-8859-1" 5, 25 -- X-OriginalArrivalTime: 18 Apr 2006 19:35:39.0734 (UTC) FILETIME=[4611B760:01C6631F] ==============================End of - Headers==============================
are the splits along a tissue-wax interface? If yes, then your infiltration isn't complete.
Are you sure that you aren't leaving any tiny bits of razor blade or other contaminants when you trim the face prior to sectioning?
Do the splits always occur at the same position ON THE KNIFE? If yes, what type of knife are you using? If its disposable, dispose of it and try a new one. If its the kind you resharpen, then it needs to be resharpened. Even very tiny flaws in the knife edge can cause scratches on the block face that translate to splits in the sections.
Can you adjust the clearance angle? The block could be rubbing against the knife holder/stage once it sweeps past the knife edge itself.
I know this sounds silly, but are you using the correct chuck holder? It happened in my lab. My technician was using the chuck holder designed for square blocks (as from Peel-Away molds) to clamp the holder for the TissueTek bases. This resulted in the block extending too far forward from the arm and rubbing against the knife stage. She was frustrated by the scratched/splits in her sections, but no one noticed the mistake until we had the microtome serviced and the service guy asked us why we were doing that!
Those are my ideas. I'm sure you'll get others. Good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 7, 24 -- From lcgould-at-med.cornell.edu Tue Apr 18 15:05:38 2006 7, 24 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 7, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3IK5cvG020895 7, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 15:05:38 -0500 7, 24 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 7, 24 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k3IK5b6p006081 7, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 16:05:37 -0400 (EDT) 7, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 7, 24 -- by mpx1.med.cornell.edu 7, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 7, 24 -- with ESMTPA id {0IXX00DCXP5B6B60-at-mpx1.med.cornell.edu} for 7, 24 -- microscopy-at-microscopy.com; Tue, 18 Apr 2006 16:05:37 -0400 (EDT) 7, 24 -- Date: Tue, 18 Apr 2006 16:00:23 -0400 7, 24 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 7, 24 -- Subject: Re: [Microscopy] LM paraffin section methods troubleshoot 7, 24 -- In-reply-to: {200604181933.k3IJX0Vs011688-at-ns.microscopy.com} 7, 24 -- Sender: lcgould-at-med.cornell.edu 7, 24 -- To: kellie.l.garner-at-monsanto.com, 7, 24 -- Microscopy Listserver {microscopy-at-microscopy.com} 7, 24 -- Message-id: {p06230912c06af35bbe9d-at-[140.251.48.23]} 7, 24 -- MIME-version: 1.0 7, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 7, 24 -- References: {200604181933.k3IJX0Vs011688-at-ns.microscopy.com} 7, 24 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.4.18.124606 ==============================End of - Headers==============================
4th Annual Short Course and Workshop on Computer-Assisted Image Analysis and Measurement
Instructor: Dr. John C. Russ
June 28 -30, 2006
University of Missouri Columbia, MO
Image processing and analysis are critical components of many fields of science and engineering. This hands-on course, mixing step-by-step exercises, teaches the fundamental principles and techniques that are essential to obtain meaningful and useful results to solve real world problems. With the small class size and extended lab times, attendees are encouraged to bring their own images for individualized instruction. Participants will receive a trial version of the Fovea Pro software (a comprehensive package of Photoshop plug-ins) including a complete manual and all images used in the course, a road map guide to image analysis, and CEU credits.
Registration deadline: May 19, 2006 Enrollment limit: 15
For further information and an application form, visit our website: www.emc.missouri.edu/works.htm Or contact Lou Ross at (573) 882-4777 or at rosslm-at-missouri.edu
-- 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============================== 8, 16 -- From RossLM-at-missouri.edu Tue Apr 18 15:35:36 2006 8, 16 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 8, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3IKZZIE031393 8, 16 -- for {microscopy-at-microscopy.com} ; Tue, 18 Apr 2006 15:35:35 -0500 8, 16 -- Received: from um-exproto9.um.umsystem.edu ([207.160.151.148]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 8, 16 -- Tue, 18 Apr 2006 15:35:34 -0500 8, 16 -- Received: from [128.206.79.116] ([128.206.78.230]) by um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft SMTPSVC(6.0.3790.1830); 8, 16 -- Tue, 18 Apr 2006 15:35:31 -0500 8, 16 -- Mime-Version: 1.0 8, 16 -- X-Sender: RossLM-at-pop.missouri.edu 8, 16 -- Message-Id: {p05200f5fc06afdecd820-at-[128.206.79.116]} 8, 16 -- Date: Tue, 18 Apr 2006 15:35:35 -0500 8, 16 -- To: microscopy-at-microscopy.com 8, 16 -- From: Lou Ross {RossLM-at-missouri.edu} 8, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 8, 16 -- X-OriginalArrivalTime: 18 Apr 2006 20:35:31.0992 (UTC) FILETIME=[A338C980:01C66327] ==============================End of - Headers==============================
I am having troubles with AnalySIS on my Quanta microscope. I would love to get in touch with Mike Bode or someone in Support at Olympus SIS to see if we can get it working again.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 3, 23 -- From hagglundk1-at-nku.edu Wed Apr 19 09:10:46 2006 3, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3JEAjJ0022870 3, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Apr 2006 09:10:46 -0500 3, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 3, 23 -- Wed, 19 Apr 2006 10:10:44 -0400 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 3, 23 -- Content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="us-ascii" 3, 23 -- Subject: analySIS contact information 3, 23 -- Date: Wed, 19 Apr 2006 10:10:43 -0400 3, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3586FB-at-mailfac1.hh.nku.edu} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: analySIS contact information 3, 23 -- Thread-Index: AcZjuwwNke2LtdYBRMu6bpKdwSrSLg== 3, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 3, 23 -- To: {microscopy-at-microscopy.com} 3, 23 -- X-OriginalArrivalTime: 19 Apr 2006 14:10:44.0544 (UTC) FILETIME=[0C718C00:01C663BB] 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3JEAjJ0022870 ==============================End of - Headers==============================
Colleagues, has anyone ever performed PCR-genotyping/analysis of tissue which was fixed by formaldehyde- and glutaraldehyde (no contact with osmium)? How do you treat / digest your fixed tissue sample? Any advice or tips are welcome! greetings, Peter Heimann
I am getting more and more microscopy-related spam in the past month or two, usually advertisements for commercial organizations I have never dealt with or ever heard of. The latest is "Microanalysis News" from Bruker AXS Microanalysis. I someone harvesting names from our list or what?
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, 32 -- From mcauliff-at-umdnj.edu Wed Apr 19 10:28:03 2006 6, 32 -- Received: from zix04.umdnj.edu (zix04.UMDNJ.EDU [130.219.34.127]) 6, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3JFS3Cn011707 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Apr 2006 10:28:03 -0500 6, 32 -- Received: from zix04.umdnj.edu (ZixVPM [127.0.0.1]) 6, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id E95C24BE59 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Apr 2006 10:28:02 -0500 (CDT) 6, 32 -- Received: from umdnj.edu (imail.UMDNJ.EDU [130.219.34.129]) 6, 32 -- by zix04.umdnj.edu (Proprietary) with ESMTP id C2A484BE32 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Apr 2006 10:28:01 -0500 (CDT) 6, 32 -- Received: from ([130.219.34.131]) 6, 32 -- by imail.umdnj.edu with ESMTP id KP-BRACD.65045910; 6, 32 -- Wed, 19 Apr 2006 11:27:33 -0400 6, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 32 -- id {0IXZ002016E6HY-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 32 -- for microscopy-at-msa.microscopy.com; Wed, 19 Apr 2006 11:27:33 -0400 (EDT) 6, 32 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 32 -- 2004)) with ESMTP id {0IXZ00IQM6XWQN-at-Polaris.umdnj.edu} for 6, 32 -- microscopy-at-msa.microscopy.com; Wed, 19 Apr 2006 11:27:33 -0400 (EDT) 6, 32 -- Date: Wed, 19 Apr 2006 11:28:28 -0400 6, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 32 -- Subject: spam on this list? 6, 32 -- To: MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 32 -- Message-id: {4446571C.9090807-at-umdnj.edu} 6, 32 -- MIME-version: 1.0 6, 32 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 32 -- Content-transfer-encoding: 7BIT 6, 32 -- X-Accept-Language: en-us, en 6, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 32 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
Geoff: I doubt Nestor's email list has been compromised. All the microscopy-related spam I get is from people who already have my address for one reason or another. The Bruker AXS people may have acquired your address when they acquired the PGT business. Or got it from some other source; it's so hard to keep track these days.
mcauliff-at-umdnj.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 Colleagues: } } I am getting more and more microscopy-related spam in the past month } or two, usually advertisements for commercial organizations I have never } dealt with or ever heard of. The latest is "Microanalysis News" from } Bruker AXS Microanalysis. I someone harvesting names from our list or what? } } Geoff } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 22 -- From r-holdford-at-ti.com Wed Apr 19 11:01:12 2006 4, 22 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3JG1B2Y029391 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 19 Apr 2006 11:01:11 -0500 4, 22 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 22 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k3JG0vBD006137; 4, 22 -- Wed, 19 Apr 2006 11:01:07 -0500 (CDT) 4, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 22 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k3JG0vTK015025; 4, 22 -- Wed, 19 Apr 2006 11:00:57 -0500 (CDT) 4, 22 -- Message-ID: {44465EB9.8080107-at-ti.com} 4, 22 -- Date: Wed, 19 Apr 2006 11:00:57 -0500 4, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 22 -- Organization: SC Packaging Development -- FA Development 4, 22 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 4, 22 -- MIME-Version: 1.0 4, 22 -- To: mcauliff-at-umdnj.edu, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Re: [Microscopy] spam on this list? 4, 22 -- References: {200604191528.k3JFSH3n012038-at-ns.microscopy.com} 4, 22 -- In-Reply-To: {200604191528.k3JFSH3n012038-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
---- Call for Papers Deadline Extended to May 5, 2006 ---
You still have time to submit an abstract for the Seeing at the Nanoscale IV International Conference, July 17-20, 2006, at the University of Pennsylvania, Philadelphia. The Abstract Submission deadline date has been extended to Friday, May 5.
The conference theme is Exploring Nanostructure Imaging, Characterization and Modification Using SPM and Related Techniques.
This event-filled, three-day conference provides an optimum forum for "scientists to speak to scientists" on a wide variety of cutting-edge nanotechnology topics, with technical sessions on:
SESSION 1: Title: Nanomechanical & Local Property Measurements Focus: Methods to measure static and dynamic nanoscale mechanical and tribological properties, including nanoindenting, scratching and nanoDMA. The session will also concentrate on molecular models and the understanding of fundamental properties in relation to the above measurement modes. Chair: Greg Meyers, Dow Chemical Company Guest Speaker: Ozgur Sahin, Harvard University
SESSION 2: Title: Visualization I: Biomolecules & Biological Processes Focus: Techniques to image cells, proteins, lipids, and tissue samples in physiologically relevant environments including high resolution imaging of static samples and visualization of dynamic events to measure inter- and intra-molecular forces. Chair: Jan Hoh, Johns Hopkins School of Medicine Guest Speaker: Alexander Malkin, Lawrence Livermore National Labs
SESSION 3: Title: Visualization II: Materials & Polymer Systems Focus: Methods to image and manipulate, from single macromolecule and functional self-assemblies to complete materials systems Chair: Sergei Magonov, Veeco Instruments Guest Speaker: Dimitri Ivanov, Institut de Chimie des Surfaces et Interfaces, France
SESSION 4: Title: Measurements of Electrical, Optical, Magnetic & Thermal Properties of Materials at the Nanoscale Focus: Materials characterization in nanometer and sub-micron scale with emphasis on electrical, optical, magnetic and thermal properties Chair: Sergei Kalinin, Oak Ridge National Laboratory Guest Speaker: Louis Brus, Columbia University
SESSION 5: Title: Instrumentation: New Tools and Techniques for Nanoscience Focus: Innovative and future developments of SPM tools and techniques, probes and sensors Chair: Ning Xi, Michigan State University Guest Speaker: Levent Degertekin, Georgia Tech
Contributed papers will be considered for either oral or poster presentation at the conference unless authors request a poster session. The session chairs will review all abstracts. Final abstracts will be posted on the conference website and printed in the conference program.
If you are interested in submitting an abstract, please see www.veeco.com/nanoconference/call_for_papers.asp for detailed submission guidelines and session-specific information.
We know this will be a very dynamic conference, and we look forward to your participation.
Questions: Contact Marlene Carlyle at mcarlyle-at-veeco.com
Veeco Instruments www.veeco.com/nanoconference
==============================Original Headers============================== 16, 20 -- From MCarlyle-at-veeco.com Wed Apr 19 11:22:01 2006 16, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.veeco.com [68.111.35.167]) 16, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3JGM0RQ007701 16, 20 -- for {Microscopy-at-Microscopy.com} ; Wed, 19 Apr 2006 11:22:01 -0500 16, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 16, 20 -- content-class: urn:content-classes:message 16, 20 -- MIME-Version: 1.0 16, 20 -- Content-Type: text/plain; 16, 20 -- charset="iso-8859-1" 16, 20 -- Subject: AFM-STM-Seeing at the Nanoscale Conference 16, 20 -- Date: Wed, 19 Apr 2006 09:22:00 -0700 16, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F42013AE7F3-at-sboexch2.int.veeco.com} 16, 20 -- X-MS-Has-Attach: 16, 20 -- X-MS-TNEF-Correlator: 16, 20 -- Thread-Topic: AFM-STM-Seeing at the Nanoscale Conference 16, 20 -- Thread-Index: AcZjzWKwuPsfUptYTomb1j+IYXg1pw== 16, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 16, 20 -- To: {Microscopy-at-Microscopy.com} 16, 20 -- Content-Transfer-Encoding: 8bit 16, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3JGM0RQ007701 ==============================End of - Headers==============================
Any opinions on databases for image collections? We have both Mac and PC users, something simple would probably be the best.
Thanks
Jon
-- I'm riding in the MS 150 Crusin' to the Coast bike ride May 13 - 14, 2006 to raise money for the National Multiple Sclerosis Society. You can go to http://www.msconnection.org and follow the links to make an ePledge if you wish.
Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 7, 21 -- From jmkrupp-at-ucsc.edu Wed Apr 19 15:05:38 2006 7, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3JK5bH2030839 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 19 Apr 2006 15:05:38 -0500 7, 21 -- Received: from ucsc.edu (cruzmail-fe1.ucsc.edu [128.114.125.5]) 7, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.6/8.13.1) with ESMTP id k3JK5NOa024994 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 19 Apr 2006 13:05:31 -0700 (PDT) 7, 21 -- Received: from [128.114.25.185] (account jmkrupp-at-ucsc.edu [128.114.25.185] verified) 7, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 7, 21 -- with ESMTPA id 53005984 for microscopy-at-microscopy.com; Wed, 19 Apr 2006 13:05:22 -0700 7, 21 -- Mime-Version: 1.0 7, 21 -- Message-Id: {p06230903c06c47fce1bd-at-[128.114.25.185]} 7, 21 -- Date: Wed, 19 Apr 2006 13:05:21 -0700 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 7, 21 -- Subject: Image database? 7, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 7, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 7, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 7, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
These are both in the $45-$65 price range. Not sure if Mac is supported by both but probably by Thumbs.
gary g.
At 01:07 PM 4/19/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both emily.wiesner-at-medecine.unige.ch as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] High background with immunohistochemistry
Question: Hi All. I was wondering whether anyone could suggest ways to reduce background staining when performing immunohistochemistry (using DAB) in rat brain tissue that has been frozen and cut on the cryostat. In general, I tend to get a light brown background when using DAB. I currently rinse tissue with PBS, block with methanol and hydrogen peroxide for 20 min, rinse with PBS. I then block tissue with 4% bovine serum albumin (BSA) in PBS for 1 hour. I then put the primary on overnight in a solution of PBS, BSA and triton. The following day I place the secondary on for 1 hour, followed by AB complex from the ABC vectastain kit for 1 hour, then DAB with hydrogen peroxide. I rinse with PBS in between each step. Suggestions welcome! Emily
I would say you first have to pinpoint the cause of the background. What if you leave out your primary, your secondary etc? What species was the primary raised in? What incubation solutions do you use? What about your washing steps? Do you get the same background if you use a different primary? Have you repeated earlier experimenst that gave only 'a light brown background'?
A lot of questions and it is not even a complete list, I know, but they need addressing before you can make a rational decision on what to change.
Jan Leunissen
Aurion - President Present Address: Costerweg 5 EM-Unit 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4797301 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://ocem.otago.ac.nz ------------------------------------------------------------------------ -------- Please support efforts to change the Japanese Government's attitude towards whaling for scientific purposes
On 20/04/2006, at 12:53 PM, emily.wiesner-at-medecine.unige.ch wrote: } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://microscopy.com/ } MicroscopyListserver/MLFormMail.html } ---------------------------------------------------------------------- } ----- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both emily.wiesner-at-medecine.unige.ch as well as the } MIcroscopy Listserver } ---------------------------------------------------------------------- } ----- } } Email: emily.wiesner-at-medecine.unige.ch } Name: Emily Camm } } Title-Subject: [Filtered] High background with immunohistochemistry } } Question: Hi All. } I was wondering whether anyone could suggest ways to reduce } background staining when performing immunohistochemistry (using } DAB) in rat brain tissue that has been frozen and cut on the } cryostat. In general, I tend to get a light brown background when } using DAB. I currently rinse tissue with PBS, block with methanol } and hydrogen peroxide for 20 min, rinse with PBS. I then block } tissue with 4% bovine serum albumin (BSA) in PBS for 1 hour. I then } put the primary on overnight in a solution of PBS, BSA and triton. } The following day I place the secondary on for 1 hour, followed by } AB complex from the ABC vectastain kit for 1 hour, then DAB with } hydrogen peroxide. I rinse with PBS in between each step. } Suggestions welcome! } Emily } } ---------------------------------------------------------------------- } -----
==============================Original Headers============================== 8, 24 -- From leunissen-at-aurion.nl Wed Apr 19 20:12:24 2006 8, 24 -- Received: from mta208-rme.xtra.co.nz (mta208-rme.xtra.co.nz [210.86.15.119]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3K1CMWY009079 8, 24 -- for {microscopy-at-microscopy.com} ; Wed, 19 Apr 2006 20:12:23 -0500 8, 24 -- Received: from mta4-rme.xtra.co.nz ([210.86.15.193]) 8, 24 -- by mta208-rme.xtra.co.nz with ESMTP 8, 24 -- id {20060420011221.LCRY9598.mta208-rme.xtra.co.nz-at-mta4-rme.xtra.co.nz} ; 8, 24 -- Thu, 20 Apr 2006 13:12:21 +1200 8, 24 -- Received: from [192.168.1.50] ([222.152.243.93]) by mta4-rme.xtra.co.nz 8, 24 -- with ESMTP 8, 24 -- id {20060420011221.KOUK8268.mta4-rme.xtra.co.nz-at-[192.168.1.50]} ; 8, 24 -- Thu, 20 Apr 2006 13:12:21 +1200 8, 24 -- In-Reply-To: {200604200053.k3K0rko2032027-at-ns.microscopy.com} 8, 24 -- References: {200604200053.k3K0rko2032027-at-ns.microscopy.com} 8, 24 -- Mime-Version: 1.0 (Apple Message framework v749.3) 8, 24 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 24 -- Message-Id: {8487F083-C74E-4BBB-96C5-07C8C352A1F8-at-aurion.nl} 8, 24 -- Cc: microscopy-at-microscopy.com 8, 24 -- Content-Transfer-Encoding: 7bit 8, 24 -- From: Jan Leunissen {leunissen-at-aurion.nl} 8, 24 -- Subject: Re: [Microscopy] viaWWW: High background with immunohistochemistry 8, 24 -- Date: Thu, 20 Apr 2006 13:12:15 +1200 8, 24 -- To: emily.wiesner-at-medecine.unige.ch 8, 24 -- X-Mailer: Apple Mail (2.749.3) ==============================End of - Headers==============================
I have a client who needs to find an anhydrous solvent in which to disperse her powdery stuff (ferrous and silicon oxide smokes, I think) that will not take up water, will not affect refractence spectra, and will not eat the Formvar on grids. This is for TEM and, perhaps, EELS.
Any ideas?
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
==============================Original Headers============================== 6, 19 -- From tina-at-pbrc.hawaii.edu Wed Apr 19 20:49:01 2006 6, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3K1n0d3019513 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 19 Apr 2006 20:49:00 -0500 6, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 6, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k3K1mvjU017695 6, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 19 Apr 2006 15:48:58 -1000 (HST) 6, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k3K1mudg017692 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 19 Apr 2006 15:48:56 -1000 (HST) 6, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 6, 19 -- Date: Wed, 19 Apr 2006 15:48:56 -1000 (HST) 6, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 6, 19 -- X-Sender: tina-at-halia 6, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 6, 19 -- Subject: Solvent that won't affect Formvar 6, 19 -- Message-ID: {Pine.GSO.4.21.0604191544590.17666-100000-at-halia} 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Hi Jon, I use Portfolio by Extensis. It is a very versitile image database program. They have mac and PC versions. The software doesn't support Zeiss .lsm file format and I would suggest that you check first if you have other non-mainstream image formats, e.g. Gatan. John.
jmkrupp-at-ucsc.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
--
********************************* C. John Runions, Ph.D. School of Biological and Molecular Sciences Oxford Brookes University Oxford, UK OX3 0BP
For me the question of ethics has nothing to do with digital imaging. If I want to demonstrate that a protein is present in the cell nucleus and unfortunately 95% of the cells show no nuclear staining, I just choose 1 cell with nuclear staining to show want I want to demonstrate. It is unethical, on paper format or on digital format, with or without photoshop treatment. Sadly it is the kind of pictures I sometimes see in "big" papers. Or, if you really can't find the right picture, you just add "data not shown", these terms are more and more used in the literature. These are cheap results, I wonder if you can still call it science if you don't even have to demonstrate what you say.
Talking about the "validity" of paper print Vs digital images, let me remind you of a clever manipulation I witnessed (no I won't give name ;-)): someone "wanted" no gold particles labeling over a certain part of the cell. When he saw one, he just sticked the letter used to recognized the cell compartment over the gold particle on the print!! No spare time lost in the dark chamber, no complex manipulation on Photoshop. You can still say that the gold particle is present on the negative, but hey let's be realistic, who will verify?
When you take a picture in TEM, your picture represents a very small part of your sample. It is all the science of the manipulator to get a global idea of the sample and try to represent it as accurately as possible with only one (or a few) image.
Ethics starts and ends in the head of the researchers. Their tools have nothing to do with it.
Regards,
Stéphane-without-a-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
If you want to use a clean solvent for inorganic specimens I would recommend using holey or ultra-thin carbon films rather than formvar.While formvar is the standard thin film for examining biological specimens, and is perfectly beam stable under wide beam illumination at lower to intermediate magnifications it is particularly unstable under convergent beam/high beam intensity conditions (such as are typically needed for core-loss EELS). The main advantage however is that carbon films are stable for a wide range of ultra-low water anhydrous solvents, my personal preference is for high purity Ethyl Ether, mainly as it is extremely volatile and seems to produce very low/no contamination build up (another problem for EELS). I'm sure everyone on this list has a personal solvent preference however.
Matthew
tina-at-pbrc.hawaii.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi, All- } } I have a client who needs to find an anhydrous solvent in which to } disperse her powdery stuff (ferrous and silicon oxide smokes, I think) } that will not take up water, will not affect refractence spectra, and will } not eat the Formvar on grids. This is for TEM and, perhaps, EELS. } } Any ideas? } } Mahalo, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } -- Dr M.Weyland, Postdoctoral Research Associate -------------------------------------------------- Department of Applied and Engineering Physics E13 Clark Hall Cornell University Ithaca NY 14853 FAX: 607 255 7658 (Mark FAO M.Weyland) TEL: 607 255 0654 --------------------------------------------------
==============================Original Headers============================== 6, 17 -- From mw275-at-cornell.edu Thu Apr 20 07:54:25 2006 6, 17 -- Received: from authusersmtp.mail.cornell.edu (granite1.mail.cornell.edu [128.253.83.141]) 6, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KCsOKY025059 6, 17 -- for {Microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 07:54:25 -0500 6, 17 -- Received: from [127.0.0.1] (rrdhcp152-250.redrover.cornell.edu [128.84.152.250]) 6, 17 -- (authenticated bits=0) 6, 17 -- by authusersmtp.mail.cornell.edu (8.13.1/8.12.10) with ESMTP id k3KCsKan027986 6, 17 -- for {Microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 08:54:24 -0400 (EDT) 6, 17 -- Message-ID: {4447848E.7070609-at-cornell.edu} 6, 17 -- Date: Thu, 20 Apr 2006 08:54:38 -0400 6, 17 -- From: Matthew Weyland {mw275-at-cornell.edu} 6, 17 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 6, 17 -- MIME-Version: 1.0 6, 17 -- To: Microscopy-at-microscopy.com 6, 17 -- Subject: Re:Solvent that won't affect Formvar 6, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Thank you for your answer. I had a look at the Apotome technology and specially I compared the images obtained with apotome and 3D deconvolution since both tehcniques give a sharper image. What striked me is that all images showing the usefulness of apotome technology are pictures of tissues or organisms. No single cell imaging for example. I wonder why. Are there limitations for single cell imaging by apotome? It is very important for me since I want to follow the entry of a substance into cells grown in monolayer (by IF or life cell imaging).
Regards,
Stephane
--- Sven.Terclavers-at-med.kuleuven.be 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 Stephane, } } Concerning the objectives, you might want to } consider buying an EC } Plan-NeoFluar 40x 1,3 oil. A great objective for } fluorescence (offers a } clear & intense image) that I even very often prefer } instead of our 100x } oil. } } The ApoTome is based on the grid projection theory } (more details can be } found on the Zeiss webpage). The difference with } Deconvolution software is } that this device (hardware) shows you immediately, } while acquiring the } image, the result. The deconvolution software will } only show you the result } after acquisition and running the program. If than } your image does not turn } out nice, you'll have to restart from the } acquisition on, whereas with the } ApoTome, you can immediately see the result and } restart if necessary. } } Hope it helps a bit! } } Best, } } Sven Terclavers } } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } } Sent: vrijdag 7 april 2006 10:58 } To: sven.terclavers-at-med.kuleuven.be } Subject: [Microscopy] Zeiss Axiovert 200M: } objectives and axiovision } questions } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } I am currently using our Zeiss axiovert 200M to } observe cell comparments (mitochondria, golgi, } endosomes,...) by fluorescence. } For this purpose, the main "usable" objectives } available are: } - 40x/0,50 LD A-Plan } - 100x/1,3 Oil EC Plan Neofluar } } The 40x LD is useful for our life cell imaging } experiments, so I don't think we should change it. } But I wonder if the 100x is really the best } solution. } I must say that I am a little bit disappointed by } the } quality of my images (I also try to improve the } preparation of the samples!). When i look to the } "images samples" on the CD which was included with } the } axiovision soft, I notice that most of the pictures } of } cells are taken with a 63x apochromat. } I searched the site of Zeiss and found an } interesting } 63x apochromat with a NA of 1.4 and corrected for } coverglass thickness (0,17). } What is your experience with the different Zeiss } objectives? } } My second question concerns the Apotome feature of } Axiovision. It seems to do the same job as 3D } deconvolution, but how does it work? Why choose one } or } the other? } } Thank you all in advance. } } Stephane-without-i } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam } protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 7, 18 -- From nizets2-at-yahoo.com Fri Apr 7 03:54:20 } 2006 } 7, 18 -- Received: from web37402.mail.mud.yahoo.com } (web37402.mail.mud.yahoo.com [209.191.87.55]) } 7, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k378sKHp011534 } 7, 18 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Apr 2006 03:54:20 } -0500 } 7, 18 -- Received: (qmail 14286 invoked by uid } 60001); 7 Apr 2006 08:54:20 } -0000 } 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; } c=nofws; } 7, 18 -- s=s1024; d=yahoo.com; } 7, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content } -Transfer-Encoding; } 7, 18 -- } b=OMweDxN77PPBKIar7kn7dRibjJrDypQNfuQV1OKdc7dEFVm651G6nJGH/gAhXPMCsfJkczLt1k } WeVV636+rSHsiLxKVmUq/yKZTH+K6Amwn9GgQqnMwE/r83yKnGl3XwuhFTCtuPpjWtQU52gcIEsB } oZCXUHXweWZgOVPgGLlkI= ; } 7, 18 -- Message-ID: } {20060407085420.14284.qmail-at-web37402.mail.mud.yahoo.com} } 7, 18 -- Received: from [80.122.101.102] by } web37402.mail.mud.yahoo.com via } HTTP; Fri, 07 Apr 2006 01:54:20 PDT } 7, 18 -- Date: Fri, 7 Apr 2006 01:54:20 -0700 (PDT) } 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 7, 18 -- Subject: Zeiss Axiovert 200M: objectives } and axiovision questions } 7, 18 -- To: microscopy-at-microscopy.com } 7, 18 -- MIME-Version: 1.0 } 7, 18 -- Content-Type: text/plain; } charset=iso-8859-1 } 7, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== } } } Disclaimer: } http://www.kuleuven.be/cwis/email_disclaimer.htm } } } ==============================Original } Headers============================== } 21, 32 -- From Sven.Terclavers-at-med.kuleuven.be Fri } Apr 7 04:44:54 2006 } 21, 32 -- Received: from } nibbel.kulnet.kuleuven.ac.be } (nibbel.kulnet.kuleuven.ac.be [134.58.240.41]) } 21, 32 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k379irfc022136 } 21, 32 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Apr 2006 04:44:54 -0500 } 21, 32 -- Received: from localhost (localhost } [127.0.0.1]) } 21, 32 -- by nibbel.kulnet.kuleuven.ac.be (Postfix) } with ESMTP id C841B4CFFF } 21, 32 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Apr 2006 11:44:52 +0200 (CEST) } 21, 32 -- Received: from smtp02.kuleuven.be } (lepidus.kulnet.kuleuven.ac.be [134.58.240.72]) } 21, 32 -- by nibbel.kulnet.kuleuven.ac.be (Postfix) } with ESMTP id 235C14CF38 } 21, 32 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Apr 2006 11:44:52 +0200 (CEST) } 21, 32 -- Received: from smtp02.kuleuven.be } (localhost.localdomain [127.0.0.1]) } 21, 32 -- by smtp02.kuleuven.be (Postfix) with } ESMTP id E1E3E2CAAF8; } 21, 32 -- Fri, 7 Apr 2006 11:44:51 +0200 (CEST) } 21, 32 -- Received: from gwydion (unknown } [134.58.245.176]) } 21, 32 -- by smtp02.kuleuven.be (Postfix) with } ESMTP id BC7842CAAF7 } 21, 32 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Apr 2006 11:44:51 +0200 (CEST) } 21, 32 -- Reply-To: } {Sven.Terclavers-at-med.kuleuven.be} } 21, 32 -- From: "Sven Terclavers" } {Sven.Terclavers-at-med.kuleuven.be} } 21, 32 -- To: {microscopy-at-microscopy.com} } 21, 32 -- Subject: RE: [Microscopy] Zeiss Axiovert } 200M: objectives and axiovision questions } 21, 32 -- Date: Fri, 7 Apr 2006 11:44:49 +0200 } 21, 32 -- Organization: VIB CTG-CMVB } === message truncated ===
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Sadly, the "I've got a picture so it must be true." syndrome of electron microscopy has been around for a very long time - don't expect it to go away very soon. Sjostrand described it very well in 1962 (Critical evaluation of ultrastructural patterns, The Interpretation of Ultrastructure, RJC Harris, ed, Academic Press, pp47-68). In short, he stated that the person with the best story is often the one believed, not necessarily the one with the best science. Now, that is an interpretation and extrapolation of what he said, with some liberties taken. But it really sums up what he said in a single sentence. This issue has caused no end of grief with people in other fields, many of whom developed the belief that EM is a field of phenomenology. While we are better today, the problem still exists.
As for your situation, do you not provide some distribution data to explain the frequency and degree of staining, and some explaination for why it is not uniform? eg, the immunogold labelling of the expressed protein is over granules in the microorganism, and where there are no granules there is no labelling.... The labelling on the rhinovirus particle indicated the antibody recognized an epitope expressed on the virion surface....
However, I would not universally attribute the 'data not shown' statement to fuzzy or incomplete data. Everytime that has been in a paper which I have participated in, the statement is the result of a reviewer suggesting removal of a figure because of the amount of material already in the paper. Also, when I've ask for information that is listed as 'data not shown', the overwhelming norm is that the author or speaker does not hesitate to provide it.
By the way, that full first paragraph of Shostrand's paper is prominantly displayed on my lab wall, with a recommendation that all students and budding scientists wanting to do EM read it.
paul
==============================Original Headers============================== 9, 21 -- From paul_hazelton-at-umanitoba.ca Thu Apr 20 09:53:23 2006 9, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KErMH2014296 9, 21 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 09:53:23 -0500 9, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 9, 21 -- (authenticated bits=0) 9, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k3KErKXI018116; 9, 21 -- Thu, 20 Apr 2006 09:53:21 -0500 (CDT) 9, 21 -- Message-ID: {4447A05B.2010004-at-umanitoba.ca} 9, 21 -- Date: Thu, 20 Apr 2006 09:53:15 -0500 9, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 9, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 9, 21 -- X-Accept-Language: en-us, en 9, 21 -- MIME-Version: 1.0 9, 21 -- To: nizets2-at-yahoo.com 9, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 9, 21 -- Subject: Re: [Microscopy] unethical digital imaging... 9, 21 -- References: {200604201250.k3KCoimQ019042-at-ns.microscopy.com} 9, 21 -- In-Reply-To: {200604201250.k3KCoimQ019042-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==============================
You are of course right: Ethical or unethical behavior happens in the brain, and you can use a tool ethically or unethically. The examples you provide are all valid, and in my opinion it is the peer review process that eventually must catch unethical behavior and try to shut it down.
As you said, a tool is neither ethical not unethical. However, a tool can be used to discourage unethical behavior. In areas where this is of higher importance (medical research, forensics), the appropriate organizations have already taken steps in that direction. The FDA, for example, mandates in their "rule 11" documents, that the original of an image must be stored. Granted, that does not prevent someone from deliberately looking for the single cell that shows a certain phenomenon and then writing a paper as if it was wide-spread, but it will allow other people to scrutinize the recorded evidence and come to their own conclusions (the researcher either has multiple images and only one shows the result, or he/she has only one image in total, both casting doubts on extrapolations from the images). That may prevent "ad-hoc" or "accidental" unethical behavior. It probably cannot prevent carefully planned and executed unethical behavior, but that is more of a moral and perhaps educational issue and less a technological.
mike
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Thursday, April 20, 2006 6:54 AM To: Mike Bode
Hi,
For me the question of ethics has nothing to do with digital imaging. If I want to demonstrate that a protein is present in the cell nucleus and unfortunately 95% of the cells show no nuclear staining, I just choose 1 cell with nuclear staining to show want I want to demonstrate. It is unethical, on paper format or on digital format, with or without photoshop treatment. Sadly it is the kind of pictures I sometimes see in "big" papers. Or, if you really can't find the right picture, you just add "data not shown", these terms are more and more used in the literature. These are cheap results, I wonder if you can still call it science if you don't even have to demonstrate what you say.
Talking about the "validity" of paper print Vs digital images, let me remind you of a clever manipulation I witnessed (no I won't give name ;-)): someone "wanted" no gold particles labeling over a certain part of the cell. When he saw one, he just sticked the letter used to recognized the cell compartment over the gold particle on the print!! No spare time lost in the dark chamber, no complex manipulation on Photoshop. You can still say that the gold particle is present on the negative, but hey let's be realistic, who will verify?
When you take a picture in TEM, your picture represents a very small part of your sample. It is all the science of the manipulator to get a global idea of the sample and try to represent it as accurately as possible with only one (or a few) image.
Ethics starts and ends in the head of the researchers. Their tools have nothing to do with it.
Regards,
Stéphane-without-a-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
I agree that ethics are normally independent of the tools used.
Over the years at various places, I have been asked to emphasize a band in a photo of a gel by burning-in in the darkroom, remove peaks from EDS spectra because "they shouldn't be there", give quantitative EDS data from samples clearly unsuitable for quantitation, etc., etc.
One of the most prevalent problematic imaging sins is often unintentional, in my opinion, but clearly not always. To illustrate, I will make up an example so contrived that nobody could possibly believe I was talking about any person in particular, because I am not.
Let's say that Researcher A brings in samples of nasal tissue complete with sensory hairs from the endangered and fierce Mongolian wombat. The hypothesis is that the protein snifrin is localized in the sensory fibers of the urban population of the wombat, but not in its rural relatives, and this protein helps the city wombat to find discarded orange peels which make up a major part of its diet.
The tissue is prepped for SEM, labelled with goat anti-snifrin, followed by anti-goat 10nm gold conjugate. We coat with carbon and pop it into the scope and, LO, the urban wombat sensory hairs are lit up like Christmas trees with backscatter images of gold! Eureka!
Unfortunately, so are the rural wombat's. Not to be deterred, Researcher A searches and searches until s/he finds a few isolated unlabelled hairs, pops off a few images and publishes triumphantly that the hypothesis is verified.
This is an extreme example, but the initial selection of images to record has its own set of ethical questions, and it doesn't matter if the methods are silver-based or digital.
Cheers,
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Thursday, April 20, 2006 7:49 AM To: Tindall, Randy D.
Hi,
For me the question of ethics has nothing to do with digital imaging. If I want to demonstrate that a protein is present in the cell nucleus and unfortunately 95% of the cells show no nuclear staining, I just choose 1 cell with nuclear staining to show want I want to demonstrate. It is unethical, on paper format or on digital format, with or without photoshop treatment. Sadly it is the kind of pictures I sometimes see in "big" papers. Or, if you really can't find the right picture, you just add "data not shown", these terms are more and more used in the literature. These are cheap results, I wonder if you can still call it science if you don't even have to demonstrate what you say.
Talking about the "validity" of paper print Vs digital images, let me remind you of a clever manipulation I witnessed (no I won't give name ;-)): someone "wanted" no gold particles labeling over a certain part of the cell. When he saw one, he just sticked the letter used to recognized the cell compartment over the gold particle on the print!! No spare time lost in the dark chamber, no complex manipulation on Photoshop. You can still say that the gold particle is present on the negative, but hey let's be realistic, who will verify?
When you take a picture in TEM, your picture represents a very small part of your sample. It is all the science of the manipulator to get a global idea of the sample and try to represent it as accurately as possible with only one (or a few) image.
Ethics starts and ends in the head of the researchers. Their tools have nothing to do with it.
Regards,
Stéphane-without-a-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
I have a follow-up question to the recent postings on imaging ethics.
As a technician in a multi-user core facility, what role does someone like me have in spotting ethically questionable behavior and pointing it out? Are we obligated to alert a researcher when they may be unconsciously prejudicing their results by selective imaging? Are we expected to report it if there is a blatant example of deliberate skewing of results? Is it okay to let it happen, if we ourselves don't actively participate in it? If a publication results from a piece of research we assisted with and the images used to support a conclusion are obviously not representative of those taken during the research, do we have an obligation to comment?
I'd be curious to find out how other labs deal with such things.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 9, 23 -- From TindallR-at-missouri.edu Thu Apr 20 11:12:31 2006 9, 23 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KGCVEp012624 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 11:12:31 -0500 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Thu, 20 Apr 2006 11:12:30 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="US-ASCII" 9, 23 -- Subject: Ethical question 9, 23 -- Date: Thu, 20 Apr 2006 11:11:39 -0500 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849F41-at-UM-EMAIL09.um.umsystem.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Ethical question 9, 23 -- thread-index: AcZklRuKX2YC3GZDRjuGFPDySAbWWw== 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 20 Apr 2006 16:12:30.0978 (UTC) FILETIME=[39D4A620:01C66495] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3KGCVEp012624 ==============================End of - Headers==============================
On Apr 19, 2006, at 6:49 PM, tina-at-pbrc.hawaii.edu wrote:
} I have a client who needs to find an anhydrous solvent in which to } disperse her powdery stuff (ferrous and silicon oxide smokes, I think) } that will not take up water, will not affect refractence spectra, and } will } not eat the Formvar on grids. This is for TEM and, perhaps, EELS. } Dear Tina, Ethanol will not eat formvar, and 95% should be OK as far as taking up water. If even that amount of water is not acceptable, I'd try n-butanol (a real chemist could tell you if 100% BuOH takes up water). I have no idea whether either of these will affect refractence spectra, or whether these spectra are to be obtained on the suspensions of the particles or on the particles themselves after the solvent has dried. I also do not know whether alkanes dissolve formvar, or, if you don't mind dealing with some nasty smells, whether some of the aromatics--substituted ones like toluene, xylene, or pyradine might be better on formvar than benzene--would work. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Thu Apr 20 12:44:41 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KHifJZ023460 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Apr 2006 12:44:41 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 1B91C34A34 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Apr 2006 10:44:41 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 565C710AF40 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Apr 2006 10:38:38 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200604200149.k3K1n7Xr019694-at-ns.microscopy.com} 4, 22 -- References: {200604200149.k3K1n7Xr019694-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {4d2805af60f013b3e31e27526a96f3a0-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Solvent that won't affect Formvar 4, 22 -- Date: Thu, 20 Apr 2006 10:48:35 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Randy, Years ago, I had an investigator come to me to get SEM images for a poster he was going to present at a meeting. He told me that it wouldn't take too long, because even though he had 3 samples, he knew exactly what he was looking for. I explained to him that we needed to examine the samples thoroughly so that the images we collected would be representative of the whole. He said something like...'I need a low, medium and high mag of the control, carrier alone and the drug treated, and I know what I want to see'. Try as I might, he would not consent to taking more than those 9 images. When he walked out of the lab, I told him that he should not mention my name on his poster or in his talk. I know I "copped out". In subsequent years, I have tried to educate people about how limited the sampling in microscopy (EM especially) and that they really must spend the time to completely examine their samples, repeat the experiments, etc. My colleagues here at WMC and at neighboring institutions (other Core Facility directors) often have chatted among ourselves about whether it falls to us to tell people how to "do" their science. We haven't arrived at any consensus. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 23 -- From lcgould-at-med.cornell.edu Thu Apr 20 12:45:12 2006 1, 23 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 1, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KHjBdd024144 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 12:45:12 -0500 1, 23 -- Received: from mpx2.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 23 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k3KHj6pI000147 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 13:45:08 -0400 (EDT) 1, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 23 -- by mpx2.med.cornell.edu 1, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 23 -- with ESMTPA id {0IY1001WO7Z55I10-at-mpx2.med.cornell.edu} for 1, 23 -- microscopy-at-microscopy.com; Thu, 20 Apr 2006 13:45:06 -0400 (EDT) 1, 23 -- Date: Thu, 20 Apr 2006 13:39:31 -0400 1, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 23 -- Subject: Re: [Microscopy] Ethical question 1, 23 -- In-reply-to: {200604201613.k3KGDJtG013932-at-ns.microscopy.com} 1, 23 -- Sender: lcgould-at-med.cornell.edu 1, 23 -- To: TindallR-at-missouri.edu, Microscopy Listserver {microscopy-at-microscopy.com} 1, 23 -- Message-id: {p06230902c06d73f773b7-at-[140.251.48.23]} 1, 23 -- MIME-version: 1.0 1, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 23 -- References: {200604201613.k3KGDJtG013932-at-ns.microscopy.com} 1, 23 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.4.20.102105 ==============================End of - Headers==============================
In my experience the response of a technician/microscopist should vary depending mostly upon the people involved. The best position to take is as an educator explaining why a particular preparation, image or set of images does or does not provide accurate and sufficient evidence for a hypothesis/statement. Fortunately, most established scientists that I have worked with responded to this information appropriately. I have seen more problems with beginning scientists such as grad students, post-docs and beginning professors trying to get a first grant. My position is to offer information to the scientist regarding ethical microscopy. If they refuse to consider the information I begin to separate myself from the work and the scientist. I was very close to becoming a "whistle blower" by reporting my observations regarding statements in a grant application and a scientific paper to a journal editor, the Dean of our school and a government funding department. However, I backed off after severing my ties to that scientist and my gentle comments to the department chairman provoked lavish praise of the scientist in question. My hope is that person will suffer from a bad "karma" though justice is usually the exception rather than the rule. My comfort comes from the conviction that the scientific truth will eventually become evident despite the efforts of individuals to twist data for their own personal aggrandisement.
TindallR-at-missouri.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a follow-up question to the recent postings on imaging ethics. } } As a technician in a multi-user core facility, what role does someone } like me have in spotting ethically questionable behavior and pointing it } out? Are we obligated to alert a researcher when they may be } unconsciously prejudicing their results by selective imaging? Are we } expected to report it if there is a blatant example of deliberate } skewing of results? Is it okay to let it happen, if we ourselves don't } actively participate in it? If a publication results from a piece of } research we assisted with and the images used to support a conclusion } are obviously not representative of those taken during the research, do } we have an obligation to comment? } } I'd be curious to find out how other labs deal with such things. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W125 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 9, 23 -- From TindallR-at-missouri.edu Thu Apr 20 11:12:31 2006 } 9, 23 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) } 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KGCVEp012624 } 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 11:12:31 -0500 } 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 9, 23 -- Thu, 20 Apr 2006 11:12:30 -0500 } 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 9, 23 -- Content-class: urn:content-classes:message } 9, 23 -- MIME-Version: 1.0 } 9, 23 -- Content-Type: text/plain; } 9, 23 -- charset="US-ASCII" } 9, 23 -- Subject: Ethical question } 9, 23 -- Date: Thu, 20 Apr 2006 11:11:39 -0500 } 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849F41-at-UM-EMAIL09.um.umsystem.edu} } 9, 23 -- X-MS-Has-Attach: } 9, 23 -- X-MS-TNEF-Correlator: } 9, 23 -- Thread-Topic: Ethical question } 9, 23 -- thread-index: AcZklRuKX2YC3GZDRjuGFPDySAbWWw== } 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 9, 23 -- To: {microscopy-at-microscopy.com} } 9, 23 -- X-OriginalArrivalTime: 20 Apr 2006 16:12:30.0978 (UTC) FILETIME=[39D4A620:01C66495] } 9, 23 -- Content-Transfer-Encoding: 8bit } 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3KGCVEp012624 } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
Electron Crystallography Workshop, UC Davis California (Aug 7-11, 2006)
We have the pleasure of announcing the opening of registration for a one-week International Workshop on Electron crystallography to be held at UC Davis, California (August 7-11, 2006).
The workshop has two major aims:
1) Training: To provide a unique forum to train skilled biochemists and electron microscopists (PhD, Post docs and beyond), in state-of-the- art 2D crystallization, electron crystallography data collection and image processing.
Topics will cover all aspects of electron crystallography, including: - Membrane protein solubilisation and crystallization - Sample preparation for electron microscopy - Cryo-EM data collection by image recording and electron diffraction - Image processing, including an introduction to the new software systems IPLT and 2dx - Data evaluation and model building
2) Advancing the technology: To bring together many of the leading groups in electron crystallography to forge innovative collaborations for new technology development.
Workshop format: The format of the workshop will be similar to the EMBO courses in that lectures will be held in the morning, practicals in the afternoon, student poster session before dinner, and science talks in the evening. However the workshop will be focusing exclusively on electron crystallography related topics.
Registration: The workshop is currently limited to 20 participants. The fee for academic participants is US$200, which includes board and lodging (twin room). Registration can be completed at http://2dx.org/ workshop before the deadline of 1 June 2006.
Further information: For further information please see http:// 2dx.org/workshop or contact Henning Stahlberg (HStahlberg-at-ucdavis.edu) or Ben Hankamer (b.hankamer-at-imb.uq.edu.au).
Ben Hankamer and Henning Stahlberg
Henning Stahlberg, Molecular & Cellular Biology, Briggs Hall 5, University of California at Davis, 1 Shields Ave., Davis, CA 95616, USA Tel: +1-530-752 8282 (office), +1-530-754 8285 (lab) Fax: +1-530-752 3085 mailto:HStahlberg-at-ucdavis.edu http://stahlberglab.ucdavis.edu AIM:HenningStahlberg-at-aim.com _____________________________________________________________
==============================Original Headers============================== 22, 16 -- From HStahlberg-at-ucdavis.edu Thu Apr 20 13:55:57 2006 22, 16 -- Received: from pop23.ucdavis.edu (pop23.ucdavis.edu [169.237.105.13]) 22, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KItvBT021330 22, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 13:55:57 -0500 22, 16 -- Received: from [169.237.214.65] ([169.237.214.65]) 22, 16 -- by pop23.ucdavis.edu (8.13.6/8.13.1/it-std-5.2.0) with ESMTP id k3KItlTQ012739; 22, 16 -- Thu, 20 Apr 2006 11:55:47 -0700 (PDT) 22, 16 -- Mime-Version: 1.0 (Apple Message framework v749.3) 22, 16 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 22, 16 -- Message-Id: {07ECC180-A7A3-4965-B48E-21094C72E25E-at-ucdavis.edu} 22, 16 -- Content-Transfer-Encoding: 7bit 22, 16 -- From: Henning Stahlberg {HStahlberg-at-ucdavis.edu} 22, 16 -- Subject: Workshop on Electron Crystallography, UC Davis, August 7-11, 2006 22, 16 -- Date: Thu, 20 Apr 2006 11:55:41 -0700 22, 16 -- To: Microscopy-at-microscopy.com 22, 16 -- X-Mailer: Apple Mail (2.749.3) ==============================End of - Headers==============================
Listers, While not wanting disagree about the importance of adequate sampling and interpretation, it is worth remembering that microscopy can be used for demonstration not investigation. This is like good old Gregor Mendel who used his peas to demonstrate particulate inheritance not to figure it out. It is reasonable to want, for example, an SEM picture of something one has been studying for years with light microscopy, and in that case just getting the right shot is exactly the right move. As long as you represent it as a visualization tool not a discovery (which Mendel apparently failed to do!), I don't see a problem. Tobias } } } Randy, } Years ago, I had an investigator come to me to get SEM images for a } poster he was going to present at a meeting. He told me that it } wouldn't take too long, because even though he had 3 samples, he knew } exactly what he was looking for. I explained to him that we needed } to examine the samples thoroughly so that the images we collected } would be representative of the whole. He said something like...'I } need a low, medium and high mag of the control, carrier alone and the } drug treated, and I know what I want to see'. Try as I might, he } would not consent to taking more than those 9 images. When he walked } out of the lab, I told him that he should not mention my name on his } poster or in his talk. I know I "copped out". In subsequent years, } I have tried to educate people about how limited the sampling in } microscopy (EM especially) and that they really must spend the time } to completely examine their samples, repeat the experiments, etc. } My colleagues here at WMC and at neighboring institutions (other } Core Facility directors) often have chatted among ourselves about } whether it falls to us to tell people how to "do" their science. We } haven't arrived at any consensus. } Lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy & Histology Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } http://www.cornellcelldevbiology.org } http://www.cornellbiochem.org } } ==============================Original Headers============================== } 1, 23 -- From lcgould-at-med.cornell.edu Thu Apr 20 12:45:12 2006 } 1, 23 -- Received: from smtp-gw1.med.cornell.edu } (smtp-gw1.med.cornell.edu [140.251.3.74]) } 1, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k3KHjBdd024144 } 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 } 12:45:12 -0500 } 1, 23 -- Received: from mpx2.med.cornell.edu } (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) } 1, 23 -- by smtp-gw1.med.cornell.edu } (Switch-3.1.8/Switch-3.1.7) with ESMTP id k3KHj6pI000147 } 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 } 13:45:08 -0400 (EDT) } 1, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu } [140.251.48.23]) } 1, 23 -- by mpx2.med.cornell.edu } 1, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built } Jan 28 2005)) } 1, 23 -- with ESMTPA id {0IY1001WO7Z55I10-at-mpx2.med.cornell.edu} for } 1, 23 -- microscopy-at-microscopy.com; Thu, 20 Apr 2006 13:45:06 -0400 (EDT) } 1, 23 -- Date: Thu, 20 Apr 2006 13:39:31 -0400 } 1, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} } 1, 23 -- Subject: Re: [Microscopy] Ethical question } 1, 23 -- In-reply-to: {200604201613.k3KGDJtG013932-at-ns.microscopy.com} } 1, 23 -- Sender: lcgould-at-med.cornell.edu } 1, 23 -- To: TindallR-at-missouri.edu, Microscopy Listserver } {microscopy-at-microscopy.com} } 1, 23 -- Message-id: {p06230902c06d73f773b7-at-[140.251.48.23]} } 1, 23 -- MIME-version: 1.0 } 1, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed } 1, 23 -- References: {200604201613.k3KGDJtG013932-at-ns.microscopy.com} } 1, 23 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, } Antispam-Data: 2006.4.20.102105 } ==============================End of - Headers==============================
Hi all I am afraid that I do want to disagree. Microscopy CAN be used for investigation. I have used both fluorescence and TEM as analytical tools. As such, great thought was needed to take unbiased samples and do experiments appropriate to the technology. Tom Pollard used the TEM to workout the kinetics of actin. I have used it to workout several biological pathways. If a user is just looking for a picture that 'shows what they already know to be true', that's fine, as long as they present the picture that way.
David
} } Listers, } While not wanting disagree about the importance of adequate } sampling and interpretation, it is worth remembering that microscopy } can be used for demonstration not investigation. This is like good } old Gregor Mendel who used his peas to demonstrate particulate } inheritance not to figure it out. It is reasonable to want, for } example, an SEM picture of something one has been studying for years } with light microscopy, and in that case just getting the right shot } is exactly the right move. As long as you represent it as a } visualization tool not a discovery (which Mendel apparently failed to } do!), I don't see a problem. } Tobias
_____________________
David Elliott Ph.D. Assistant Professor Department of Cell Biology and Anatomy University of Arizona College of Medicine PO Box 245004 Tucson, AZ 85724
Voice: 520-626-7870 Fax: 520-626-2097
==============================Original Headers============================== 15, 22 -- From Elliott-at-arizona.edu Thu Apr 20 15:20:39 2006 15, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 15, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KKKdNV009917 15, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 20 Apr 2006 15:20:39 -0500 15, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 15, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 66A4EDD6DC0 15, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 20 Apr 2006 13:20:39 -0700 (MST) 15, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 15, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 5F95FDDE034 15, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 20 Apr 2006 13:20:38 -0700 (MST) 15, 22 -- Mime-Version: 1.0 (Apple Message framework v749.3) 15, 22 -- In-Reply-To: {200604201901.k3KJ1EqM003295-at-ns.microscopy.com} 15, 22 -- References: {200604201901.k3KJ1EqM003295-at-ns.microscopy.com} 15, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 15, 22 -- Message-Id: {ED9364B8-D7F3-400E-9E6F-263730160011-at-arizona.edu} 15, 22 -- Content-Transfer-Encoding: 7bit 15, 22 -- From: David Elliott {Elliott-at-arizona.edu} 15, 22 -- Subject: Re: [Microscopy] Re: Ethical question 15, 22 -- Date: Thu, 20 Apr 2006 13:20:37 -0700 15, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 15, 22 -- X-Mailer: Apple Mail (2.749.3) 15, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Listers, Oops!! Serious failure to proofread. My apologies. I meant that in some applications, the point is demonstration, while in others it is investigation. I did not at all mean that microscopy cannot be used to investigate. Sorry sorry sorry again for the confusion.
Tobias } } Hi all } I am afraid that I do want to disagree. Microscopy CAN be used for } investigation. I have used both fluorescence and TEM as analytical } tools. As such, great thought was needed to take unbiased samples } and do experiments appropriate to the technology. Tom Pollard used } the TEM to workout the kinetics of actin. I have used it to workout } several biological pathways. } If a user is just looking for a picture that 'shows what they already } know to be true', that's fine, as long as they present the picture } that way. } } David } } } } } } Listers, } } While not wanting disagree about the importance of adequate } } sampling and interpretation, it is worth remembering that microscopy } } can be used for demonstration not investigation. This is like good } } old Gregor Mendel who used his peas to demonstrate particulate } } inheritance not to figure it out. It is reasonable to want, for } } example, an SEM picture of something one has been studying for years } } with light microscopy, and in that case just getting the right shot } } is exactly the right move. As long as you represent it as a } } visualization tool not a discovery (which Mendel apparently failed to } } do!), I don't see a problem. } } Tobias } } } } } } } } _____________________ } } David Elliott Ph.D. } Assistant Professor } Department of Cell Biology and Anatomy } University of Arizona College of Medicine } PO Box 245004 } Tucson, AZ 85724 } } Voice: 520-626-7870 } Fax: 520-626-2097 } } } ==============================Original Headers============================== } 15, 22 -- From Elliott-at-arizona.edu Thu Apr 20 15:20:39 2006 } 15, 22 -- Received: from smtpgate.email.arizona.edu } (deagol.email.Arizona.EDU [128.196.133.142]) } 15, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k3KKKdNV009917 } 15, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 20 Apr 2006 } 15:20:39 -0500 } 15, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) } 15, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id } 66A4EDD6DC0 } 15, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 20 Apr 2006 } 13:20:39 -0700 (MST) } 15, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) } 15, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id } 5F95FDDE034 } 15, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 20 Apr 2006 } 13:20:38 -0700 (MST) } 15, 22 -- Mime-Version: 1.0 (Apple Message framework v749.3) } 15, 22 -- In-Reply-To: {200604201901.k3KJ1EqM003295-at-ns.microscopy.com} } 15, 22 -- References: {200604201901.k3KJ1EqM003295-at-ns.microscopy.com} } 15, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 15, 22 -- Message-Id: {ED9364B8-D7F3-400E-9E6F-263730160011-at-arizona.edu} } 15, 22 -- Content-Transfer-Encoding: 7bit } 15, 22 -- From: David Elliott {Elliott-at-arizona.edu} } 15, 22 -- Subject: Re: [Microscopy] Re: Ethical question } 15, 22 -- Date: Thu, 20 Apr 2006 13:20:37 -0700 } 15, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} } 15, 22 -- X-Mailer: Apple Mail (2.749.3) } 15, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu } ==============================End of - Headers==============================
I also disagree with Tobias. You could use the same argument about any tool used in research. If one does an experiment and uses one (or more) tools to demonstrate the results of that experiment, you have done an investigation. And how could Mendel demonstrate inheritance but not figure it out?
Geoff
baskin-at-bio.umass.edu wrote:
} Listers, } While not wanting disagree about the importance of adequate } sampling and interpretation, it is worth remembering that microscopy } can be used for demonstration not investigation. This is like good } old Gregor Mendel who used his peas to demonstrate particulate } inheritance not to figure it out. It is reasonable to want, for } example, an SEM picture of something one has been studying for years } with light microscopy, and in that case just getting the right shot } is exactly the right move. As long as you represent it as a } visualization tool not a discovery (which Mendel apparently failed to } do!), I don't see a problem. } Tobias } } -- -- ********************************************** 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, 34 -- From mcauliff-at-umdnj.edu Thu Apr 20 16:23:07 2006 6, 34 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 6, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KLN7AX029893 6, 34 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Apr 2006 16:23:07 -0500 6, 34 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 6, 34 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 372A04BE51 6, 34 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Apr 2006 16:23:07 -0500 (CDT) 6, 34 -- Received: from umdnj.edu (imail.UMDNJ.EDU [130.219.34.129]) 6, 34 -- by zix02.umdnj.edu (Proprietary) with ESMTP id 903344BE59 6, 34 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Apr 2006 16:23:05 -0500 (CDT) 6, 34 -- Received: from ([130.219.34.131]) 6, 34 -- by imail.umdnj.edu with ESMTP id KP-BRACD.65265282; 6, 34 -- Thu, 20 Apr 2006 17:22:48 -0400 6, 34 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 34 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 34 -- id {0IY100K01HZCGA-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 34 -- for microscopy-at-msa.microscopy.com; Thu, 20 Apr 2006 17:22:48 -0400 (EDT) 6, 34 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 34 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 34 -- 2004)) with ESMTP id {0IY1002O1I20CO-at-Polaris.umdnj.edu} ; Thu, 6, 34 -- 20 Apr 2006 17:22:48 -0400 (EDT) 6, 34 -- Date: Thu, 20 Apr 2006 17:23:43 -0400 6, 34 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 34 -- Subject: Re: [Microscopy] Re: Ethical question 6, 34 -- In-reply-to: {200604201859.k3KIxhX5029605-at-ns.microscopy.com} 6, 34 -- To: baskin-at-bio.umass.edu, MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 34 -- Message-id: {4447FBDF.4040605-at-umdnj.edu} 6, 34 -- MIME-version: 1.0 6, 34 -- Content-type: text/plain; format=flowed; charset=us-ascii 6, 34 -- Content-transfer-encoding: 7BIT 6, 34 -- X-Accept-Language: en-us, en 6, 34 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 34 -- Gecko/20040804 Netscape/7.2 (ax) 6, 34 -- References: {200604201859.k3KIxhX5029605-at-ns.microscopy.com} ==============================End of - Headers==============================
Geoff, Hopefully you saw my correction about what I meant to say.
As for Mendel, at the time everyone thought that inheritance was blending, like mixing paint colors. But this puzzled breeders who knew that things could unmix surprisingly in successive generations (what we might call an F2). Mendel had the idea of particulate inheritance and did the math. It explained this un-mixing perfectly. And he set up his pea plants to show this. In fact, there is a subtle statistical feature that he missed completely, RA Fisher spotted it, which shows that the data in his paper were fudged. This had to do with something in an F3 or F4 where a lot of plants have to be counted and according to Fisher it is overwhelming unlikely that he could have actually obtained the results he reported, but those are exactly the numbers he expected. This example indeed shows how demonstration is perhaps a second order to investigation; but on the other hand, no one would deny that Mendel's idea was pretty important and warranted a show.
Tobias
} I also disagree with Tobias. You could use the same argument about } any tool used in research. If one does an experiment and uses one } (or more) tools to demonstrate the results of that experiment, you } have done an investigation. And how could Mendel demonstrate } inheritance but not figure it out? } } Geoff } } baskin-at-bio.umass.edu wrote: } } } Listers, } } While not wanting disagree about the importance of adequate } } sampling and interpretation, it is worth remembering that } } microscopy can be used for demonstration not investigation. This is } } like good old Gregor Mendel who used his peas to demonstrate } } particulate inheritance not to figure it out. It is reasonable to } } want, for example, an SEM picture of something one has been } } studying for years with light microscopy, and in that case just } } getting the right shot is exactly the right move. As long as you } } represent it as a visualization tool not a discovery (which Mendel } } apparently failed to do!), I don't see a problem. } } Tobias } } } -- } -- } ********************************************** } 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 } **********************************************
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both slc6-at-lehigh.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: slc6-at-lehigh.edu Name: Sharon Coe
Organization: Lehigh University
Title-Subject: [Filtered] Lehigh Microscopy School
Question: There is still time to register for the Lehigh Microscopy School (Lehigh University, Bethlehem, PA). The courses being offered in June 2006 are:
SEM and X-ray Microanalysis (June 5-9)
New Operator Introduction (June 4)
Problem Solving with SEM and X-ray Microanalysis (June 12-16)
Quantitative X-ray Microanalysis of Bulk Specimens and Particles (June 12-16)
Analytical Electron Microscopy at the Nanometer-Scale (June 12-15)
Focused Ion Beam (FIB) Instrumentation and Applications (June 12-15)
Scanning Probe Microscopy: } From Fundamentals to Advanced Applications (June 5-8)
The Lehigh Microscopy School has a 36-year record of excellence!
For more information please contact Sharon Coe sharon.coe-at-lehigh.edu 610-758-5133 www.lehigh.edu/microscopy
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both chakravarthi-at-ccmb.res.in as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: What is the area covered by the Microscope objective(like 10x, 20x etc..) at the sample? Any formulae is avialable to calculate the area covered by the objective.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both Peadarpiper-at-aol.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: Peadarpiper-at-aol.com Name: Peadar Piper
Title-Subject: [Filtered] Hitachi S-4500 Bake-Out
Question: I have a Hitachi 4500 which I need to bake out because it has not been used for a couple of years. I have all of the required heater elements etc but have been unable to figure out exactly what to do with them as it has never been a requirement on my other W filament SEM. I would be grateful if anyone could pass on any information on the procedure that should be used. I have tried contacting Hisco in the UK but have not received any response. Thanks in advance.
This Question was submitted to Ask-A-Microscopist by (wickram-at-engr.colostate.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, April 17, 2006 at 10:07:56 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both wickram-at-engr.colostate.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Title: SEM images of regenerated cellulose ultrafiltration membranes
Question: We are tryign to take SEM images of 30 to 500 kDa ultrafiltration membranes. The Ultrafiltration layer is regenerated cellulose. The problem we have is that drying the membrane leads to collapse of the pores. Is there a method e.g. use of super critical carbon dioxide, that cab be used to prevent collapse of the pores when the membranes are dried?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ppereira-at-ibili.uc.pt) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, April 20, 2006 at 06:54:55 ---------------------------------------------------------------------------
Email: ppereira-at-ibili.uc.pt Name: Paulo Pereira
Organization: Faculty of Medicine - University of Coimbra
Education: Graduate College
Location: Coimbra, Portugal
Question: Hi everybody, I am experiencing a dilemma. I have to buy a TEM for applications in cell biology at the Faculty of Medicine. At this point I have to choose between the Tecnai BioTwin (FEI) equipped with a SIS megaview camera (res 1392 x 1040) and JEM-1230 (JEOL) equipped with a Gatan camera (res 1350 x 1040). The two models are similar in terms of technical characteristics and price. Can anyone help with comments or suggestions? Many thanks
What a good dilemma to have! Whatever your choice, it can't be "wrong".
Given that both the FEI and JEOL products are of the highest quality, and will fully meet your needs, then I would start to look at secondary issues:
Service: Where is the service engineer based? Where is the parts depot (i.e. if the engineer needs parts, where will they be sent from)? How much will the service contract cost (assuming you will buy a contract, after the first year of warranty). If you will not buy a contract, what service arrangements will you make?
Training: Are there other instruments that you or your colleagues use? It there any advantage in having a similar one?
Good luck with your purchase, and I'm sure you will enjoy whichever one you choose!
Tony.
At 09:34 AM 4/21/2006, you wrote:
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ppereira-at-ibili.uc.pt) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Thursday, April 20, 2006 at 06:54:55 } --------------------------------------------------------------------------- } } Email: ppereira-at-ibili.uc.pt } Name: Paulo Pereira } } Organization: Faculty of Medicine - University of Coimbra } } Education: Graduate College } } Location: Coimbra, Portugal } } Question: Hi everybody, I am experiencing a dilemma. I have to buy a } TEM for applications in cell biology at the Faculty of Medicine. At } this point I have to choose between the Tecnai BioTwin (FEI) } equipped with a SIS megaview camera (res 1392 x 1040) and JEM-1230 } (JEOL) equipped with a Gatan camera (res 1350 x 1040). The two } models are similar in terms of technical characteristics and price. } Can anyone help with comments or suggestions? Many thanks
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Question: What is the area covered by the Microscope objective(like 10x, 20x etc..) at the sample? Any formulae is available to calculate the area covered by the objective.
Good question...
I guess the easy answer is to measure it.. Assume a circular field, use a stage micrometer and calculate the area of a circle of known diameter.
But to calculate it you need (I suspect) the working distance and numerical aperture to calculate the base area of a cone. NA is sine of (angular aperture/2). This would give you the angle and the working distance the height of the cone.... OH.. Just focus on a specimen, close the condenser diaphragm to the very edge of the field useing the back focal plane. You can remove the condenser (while not disturbing the diaphragm and measure the opening. Don't use the flip up lens in this experiment.
Second OH.. I found a formula in Micro Memo published by Wild with the formula: actual field of view D= S/(q*Vob)
I'm sure that helps... D= diameter S= field number q= tube factor and Vob= Magnification of objective
Yes, some of my scope still have draw tubes....... On additionalreflection of that formula, just measure it...
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
chakravarthi-at-ccmb .res.in To: frank.karl-at-degussa.com cc: 04/21/2006 09:25 Subject: [Microscopy] viaWWW: optical Microscopy AM Please respond to chakravarthi
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both chakravarthi-at-ccmb.res.in as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: What is the area covered by the Microscope objective(like 10x, 20x etc..) at the sample? Any formulae is avialable to calculate the area covered by the objective.
I have seen two replies this morning to questions that were posted via the Ask-a-microscopist web page. Neither of them CC-ed the poster of the original question.
Please note the following lines which are part of the standard boilerplate that gets prepended to these outside queries. ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both chakravarthi-at-ccmb.res.in as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Warren
==============================Original Headers============================== 4, 29 -- From wesaia-at-iastate.edu Fri Apr 21 09:37:12 2006 4, 29 -- Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) 4, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3LEbCB2002323 4, 29 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 21 Apr 2006 09:37:12 -0500 4, 29 -- Received: from mailout-1.iastate.edu (mailout-1.iastate.edu [129.186.140.1]) 4, 29 -- by mailhub-3.iastate.edu (8.12.11.20060308/8.12.10) with SMTP id k3LEbCSW006662 4, 29 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 21 Apr 2006 09:37:12 -0500 4, 29 -- Received: from owa.eng.iastate.edu(129.186.23.85) by mailout-1.iastate.edu via smtp 4, 29 -- id 56c7_c104d7ca_d144_11da_94d0_00304811d932; 4, 29 -- Fri, 21 Apr 2006 14:40:18 +0000 4, 29 -- Received: from maire.eng.iastate.edu ([10.10.196.69]) by owa.eng.iastate.edu with Microsoft SMTPSVC(6.0.3790.1830); 4, 29 -- Fri, 21 Apr 2006 09:37:12 -0500 4, 29 -- Content-class: urn:content-classes:message 4, 29 -- MIME-Version: 1.0 4, 29 -- Content-Type: text/plain; 4, 29 -- charset="US-ASCII" 4, 29 -- Subject: Remember to copy the sender 4, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 29 -- Date: Fri, 21 Apr 2006 09:38:19 -0500 4, 29 -- Message-ID: {16A330AC32056A40B32842EC4BB8D727AAA2BF-at-maire.eng.iastate.edu} 4, 29 -- X-MS-Has-Attach: 4, 29 -- X-MS-TNEF-Correlator: 4, 29 -- Thread-Topic: Remember to copy the sender 4, 29 -- Thread-Index: AcZlUTuU1+6/gcybQo2fCHkt5WV6Tw== 4, 29 -- From: "Straszheim, Warren E [CCE E]" {wesaia-at-iastate.edu} 4, 29 -- To: "MSA" {Microscopy-at-msa.microscopy.com} 4, 29 -- X-OriginalArrivalTime: 21 Apr 2006 14:37:12.0359 (UTC) FILETIME=[13AE6370:01C66551] 4, 29 -- Content-Transfer-Encoding: 8bit 4, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3LEbCB2002323 ==============================End of - Headers==============================
My colleague Dr. Art Miller at NIOSH in Spokane, WA, has this book to give away to a good home. Just reply to him directly and make your arrangement for delivery. Obviouosly, its a first come first taker kind of deal.
} very new hardbound book called } Handbook of Modern ion Beam Analysis, by Tesmer and Nastasi
Send your query to Art at:
aom0-at-cdc.gov
Please do not reply to me or this Listserver. -- Gilbert (Gib) Ahlstrand, Electron Microscopist, Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
==============================Original Headers============================== 6, 18 -- From ahlst007-at-umn.edu Fri Apr 21 09:47:50 2006 6, 18 -- Received: from mtaout-w.tc.umn.edu (mtaout-w.tc.umn.edu [160.94.160.21]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3LElodr012177 6, 18 -- for {Microscopy-at-Microscopy.com} ; Fri, 21 Apr 2006 09:47:50 -0500 6, 18 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) by mtaout-w.tc.umn.edu with ESMTP; Fri, 21 Apr 2006 09:47:34 -0500 (CDT) 6, 18 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 6, 18 -- Message-ID: {4448F05E.6070405-at-umn.edu} 6, 18 -- Date: Fri, 21 Apr 2006 09:46:54 -0500 6, 18 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 6, 18 -- Reply-To: ahlst007-at-umn.edu 6, 18 -- Organization: Imaging Center UM 6, 18 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 6, 18 -- X-Accept-Language: en-us, en 6, 18 -- MIME-Version: 1.0 6, 18 -- To: Microscopy-at-Microscopy.com 6, 18 -- Subject: Ion beam analysis reference book 6, 18 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
The formula is a very simple one. The first piece of information that you need is the "field number". This is a value inscribed on the EYEPIECES. It is the diameter of the field that just the eyepieces see, measured in mm; it will range between 18 and 32mm.
The formula is: Field of view = field number/Magnification of objective*
In the simplest case, if you have a field number of 22mm and a 10x objective, the diameter of the field of view will be 2.2mm or 2,200micrometers.
There is one caveat (see * above) : if there is any sort of intermediate magnification (ex: on the Zeiss systems, an "optivar" will allow you to dial in a variety of intermediate lenses providing something like 0.5x, 1x, 1.2x and 1.6x or 2x) OR if there is a tube lens with magnification other than 1 (look below the binocular/trinocular body on the intermediate piece), you must multiply the magnification of the objective times that number and use the resulting value.
So, if you have the case above, but you have a 2x tube lense or intermediate magnification, then the field of view would be 22mm/20x or 1,100 micrometers.
Hope this is 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 September. 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 08:27 AM 4/21/2006, chakravarthi-at-ccmb.res.in wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
How big a territory do you need to image and how big a difference in topography is there on your membrane? This may be another place that AFM can help. We can run these samples in a liquid cell to maintain the moisture content, if the differences in topography are not too great. Based on some general polymer studies we've done, I would think that a phase image would be very revealing.
If you would like us to run a test sample, please contact me off line.
Hope this is 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 September. 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 08:28 AM 4/21/2006, wickram-at-engr.colostate.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 15, 19 -- From bfoster-at-mme1.com Fri Apr 21 14:11:33 2006 15, 19 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 15, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3LJBXkI029619 15, 19 -- for {microscopy-at-microscopy.com} ; Fri, 21 Apr 2006 14:11:33 -0500 15, 19 -- Received: (qmail 1391 invoked by uid 2020); 21 Apr 2006 15:12:54 -0400 15, 19 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 15, 19 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 21 Apr 2006 15:12:54 -0400 15, 19 -- Message-Id: {7.0.1.0.0.20060421140904.020d9c18-at-mme1.com} 15, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 19 -- Date: Fri, 21 Apr 2006 14:11:42 -0500 15, 19 -- To: wickram-at-engr.colostate.edu, 15, 19 -- microscopy Listserver {microscopy-at-microscopy.com} 15, 19 -- From: Barbara Foster {bfoster-at-mme1.com} 15, 19 -- Subject: Re: [Microscopy] AskAMicroscopist: SEM images of regenerated 15, 19 -- cellulose 15, 19 -- In-Reply-To: {200604211328.k3LDSiMl020549-at-ns.microscopy.com} 15, 19 -- References: {200604211328.k3LDSiMl020549-at-ns.microscopy.com} 15, 19 -- Mime-Version: 1.0 15, 19 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Ranil Wickramasinghe wrote: ===================================================== Title: SEM images of regenerated cellulose ultrafiltration membranes
Question: We are tryign to take SEM images of 30 to 500 kDa ultrafiltration membranes. The Ultrafiltration layer is regenerated cellulose. The problem we have is that drying the membrane leads to collapse of the pores. Is there a method e.g. use of super critical carbon dioxide, that cab be used to prevent collapse of the pores when the membranes are dried? =========================================================== We have critical point dried ultra filtration membranes (but maybe not cellulose), however the pores in the top skin are still very difficult to resolve even by TEM because of insufficient contrast. By SEM, we have been able to obtain images only of the support structure but not of any pores in the outer skin. This is much more of a TEM than SEM application.
Some years ago we had a system in which we precipitated silver chloride into the pores, did a low acid GMA embedding (to avoid an alcohol dehydration step), cryo thin sectioning, and thought we had decorated the pores. But that to me is the only way one could really resolve the pores in the top skin. I have no idea if this approach could work for your system, you would have to just try it and find out.
Disclaimer: SPI Supplies conducts such studies through our laboratory services division, Structure Probe.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 10, 14 -- From cgarber-at-2spi.com Sun Apr 23 01:48:57 2006 10, 14 -- Received: from rwcrmhc12.comcast.net (rwcrmhc12.comcast.net [204.127.192.82]) 10, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3N6mviY011846 10, 14 -- for {Microscopy-at-msa.microscopy.com} ; Sun, 23 Apr 2006 01:48:57 -0500 10, 14 -- Message-Id: {200604230648.k3N6mviY011846-at-ns.microscopy.com} 10, 14 -- Received: from [127.0.0.1] (unknown[71.225.86.11](misconfigured sender)) 10, 14 -- by comcast.net (rwcrmhc12) with SMTP 10, 14 -- id {20060423064856m1200coanqe} ; Sun, 23 Apr 2006 06:48:56 +0000 10, 14 -- To: MICROSCOPY BB {Microscopy-at-msa.microscopy.com} 10, 14 -- Subject: Imaging of pores in ultrafiltration membrane 10, 14 -- Date: Sun, 23 Apr 2006 02:48:53 -0500 10, 14 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 10, 14 -- X-Mailer: E-Mail Connection v3.1a 10, 14 -- CC: "wickram-at-engr.colostate.edu" {wickram-at-engr.colostate.edu} ==============================End of - Headers==============================
Background: I have attempted (with limited success) to visualise the subcellular trafficking route of a protein toxin conjugated to peroxidase (POD- Toxin) by way of diaminobenzidine (DAB) and TEM. The method I use is an adaptation of similar work done by Sandvig et al. (1992) and van Deurs, et al. (1986) whom have both used DAB and TEM to visualise the trafficking routes of POD-conjugated Shiga and ricin toxins, respectively in eukaryotic cells. For those unfamiliar with their methods, I've included a brief method at the end with references.
Problems: 1. Samples (including negative controls *not* exposed to POD-toxin) contain a significant amount of unwanted "dark fluff" when viewed using a TEM (Philips CM100). The fluff looks like slivers of electron dense material (or a chemical precipitate?) of approx. 40nm in length and appear both on the cell surface and inside the cell. Strangely, the fluff seems to be cell-specific; some cells harbour the fluff while adjacent cells are comcpletely clean. Any advice on producing "fluff free" specimens would be greatly appreciated. Note that I can forward pictures of this to those interested.
2. Visualisation of the POD-toxin conjugate is proving difficult with a weak signal. Is this just a matter of reacting with DAB for longer or is there something missing from my method below?
Any help on either reducing fluff and increasing the POD-toxin signal in my samples would be greatly appreciated.
Regards, Damien Chong
PS. The method in brief: - Adherent cells grown in a flask were exposed to POD-conjugated toxin for desired length of time (1 - 2 hr -at- 37degC) - Fixed with 2.5% gluteraldehyde (60min), then washed with PBS - Reacted with 2ml PBS containing 1mg DAB and 2microL 15% H2O2 (30min) - Detached cells with rubber policeman/scraper and pelleted (25min -at- 1600 x g) - Postfixed with 2% OsO4 in water (60min -at- 4degC) - Stained with 1% uranyl acetate in water (60min -at- room temp) - Dehydrated in graded series of EtOH and embedded in resin
References: Sandvig, K., O. Garred, K. Prydz, J. V. Kozlov, S. H. Hansen, and B. van Deurs. 1992. Retrograde transport of endocytosed Shiga toxin to the endoplasmic reticulum. Nature 358:510-2.
van Deurs, B., T. I. Tonnessen, O. W. Petersen, K. Sandvig, and S. Olsnes. 1986. Routing of internalized ricin and ricin conjugates to the Golgi complex. J Cell Biol 102:37-47.
Damien Chong Microbiology & Immunology Molecular Life Sciences Building The University of Adelaide
==============================Original Headers============================== 9, 26 -- From damien.chong-at-adelaide.edu.au Sun Apr 23 23:06:53 2006 9, 26 -- Received: from odie.services.adelaide.edu.au (pulteney-pix.border.net.adelaide.edu.au [192.43.227.18]) 9, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3O46pFt023362 9, 26 -- for {Microscopy-at-Microscopy.Com} ; Sun, 23 Apr 2006 23:06:52 -0500 9, 26 -- Received: from stimpy.services.adelaide.edu.au (stimpy.services.adelaide.edu.au [10.0.10.10]) 9, 26 -- by odie.services.adelaide.edu.au (8.13.1/8.13.1) with ESMTP id k3O46pac014402 9, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 24 Apr 2006 13:36:51 +0930 9, 26 -- Received: from stimpy.services.adelaide.edu.au (localhost.localdomain [127.0.0.1]) 9, 26 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1) with ESMTP id k3O46pYD027610 9, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 24 Apr 2006 13:36:51 +0930 9, 26 -- Received: (from apache-at-localhost) 9, 26 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1/Submit) id k3O46pnW027608 9, 26 -- for Microscopy-at-Microscopy.Com; Mon, 24 Apr 2006 13:36:51 +0930 9, 26 -- Received: from molb00325.molecularbiosciences.adelaide.edu.au (molb00325.molecularbiosciences.adelaide.edu.au [129.127.106.33]) 9, 26 -- by webmail.adelaide.edu.au (IMP) with HTTP 9, 26 -- for {dchong01-at-localhost} ; Mon, 24 Apr 2006 13:36:51 +0930 9, 26 -- Message-ID: {1145851611.444c4edb23ad8-at-webmail.adelaide.edu.au} 9, 26 -- Date: Mon, 24 Apr 2006 13:36:51 +0930 9, 26 -- From: Damien Chong {damien.chong-at-adelaide.edu.au} 9, 26 -- To: Microscopy-at-Microscopy.Com 9, 26 -- Subject: TEM. Peroxidase-DAB for sub-cellular protein tracking 9, 26 -- MIME-Version: 1.0 9, 26 -- Content-Type: text/plain; charset=ISO-8859-1 9, 26 -- Content-Transfer-Encoding: 8bit 9, 26 -- User-Agent: Internet Messaging Program (IMP) 3.2.6 9, 26 -- X-Originating-IP: 129.127.106.33 ==============================End of - Headers==============================
The question you raised is a good one, and one which is largely ignored today. It seems we are somehow expected to know this stuff intuitively, or to absorb it during the mentoring process which (should) constitute graduate education. I have thought about these things quite a bit during the 12 years I spent doing construction work following a string of postdocs in which I witnessed so much scientific misconduct that I lost faith in the literature. The coup de grace was a micrograph which others claimed to represent a pure preparation of membrane vesicles, but which turned out (18 months later when I tracked down the person who took the picture) to be just a selected field from what was in reality a bunch of heterogeneous glop. I suddenly realized why I couldn't reproduce the results of the others in the lab. Then I figured out how two of the others had deceived themselves with help from a GIGO statistical analysis protocol (random numbers return a lineweaver-burke R2 of .98!), fear of not getting tenure, and a grad student who obediently followed his advisor's admonishment "I'm tired of experiments not working. I want the experiments to start working." Upon consulting, confidentially, with several senior faculty and administrators, I was advised "Don't poison the well", and "Just vote with your feet". So I became a carpenter. BTW, I now do my science as an amateur, and study things which aren't dependent upon the validity of others' work. (Yes, it's quite possible to publish from your home address!).
In a nutshell, everyone's gig is different; nobody can tell you what to do in a given situation. I adhered to the Yiddish proverb "Tell the truth and run.", but I was a street-smart kid who had skills to fall back on, and I have known many who haven't had that luxury. My best advice: KEEP A METICULOUS LAB NOTEBOOK. My mentor, bless her, demanded that I be most punctilious in my data recording, something I have found to my dismay is all too rare today. I would be happy to correspond with you offlist to share my experiences and thoughts in these matters.
Paul Grover, Ph.D. Grover Roofing and Remodeling
Randy Tindall wrote:
---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
I have a follow-up question to the recent postings on imaging ethics.
As a technician in a multi-user core facility, what role does someone like me have in spotting ethically questionable behavior and pointing it out? Are we obligated to alert a researcher when they may be unconsciously prejudicing their results by selective imaging? Are we expected to report it if there is a blatant example of deliberate skewing of results? Is it okay to let it happen, if we ourselves don't actively participate in it? If a publication results from a piece of research we assisted with and the images used to support a conclusion are obviously not representative of those taken during the research, do we have an obligation to comment?
I'd be curious to find out how other labs deal with such things.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 9, 23 -- From TindallR-at-missouri.edu Thu Apr 20 11:12:31 2006 9, 23 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3KGCVEp012624 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 11:12:31 -0500 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Thu, 20 Apr 2006 11:12:30 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="US-ASCII" 9, 23 -- Subject: Ethical question 9, 23 -- Date: Thu, 20 Apr 2006 11:11:39 -0500 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849F41-at-UM-EMAIL09.um.umsystem.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Ethical question 9, 23 -- thread-index: AcZklRuKX2YC3GZDRjuGFPDySAbWWw== 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 20 Apr 2006 16:12:30.0978 (UTC) FILETIME=[39D4A620:01C66495] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3KGCVEp012624 ==============================End of - Headers==============================
--------------------------------------------------------------- In every object there is inexhaustible meaning; the eye sees in it what the eye brings means of seeing. - Thomas Carlyle
==============================Original Headers============================== 22, 21 -- From pgrover-at-bilbo.bio.purdue.edu Mon Apr 24 12:28:22 2006 22, 21 -- Received: from mailhub128.itcs.purdue.edu (mailhub128.itcs.purdue.edu [128.210.5.128]) 22, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3OHSMX8015525 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 Apr 2006 12:28:22 -0500 22, 21 -- Received: from paklabpgrover (dhcp155-122.bio.purdue.edu [128.210.155.122]) 22, 21 -- by mailhub128.itcs.purdue.edu (8.13.6/8.13.4/internal-smtp) with ESMTP id k3OHSLCP010642 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 Apr 2006 13:28:21 -0400 22, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} 22, 21 -- To: {microscopy-at-microscopy.com} 22, 21 -- Subject: re: ethical question 22, 21 -- Date: Mon, 24 Apr 2006 13:28:26 -0400 22, 21 -- Message-ID: {000001c667c4$7eb3a070$7a9bd280-at-paklabpgrover} 22, 21 -- MIME-Version: 1.0 22, 21 -- Content-Type: text/plain; 22, 21 -- charset="us-ascii" 22, 21 -- Content-Transfer-Encoding: 7bit 22, 21 -- X-Mailer: Microsoft Office Outlook 11 22, 21 -- Thread-Index: AcZnxH5btnNRbsYCThmNsNFgawelDA== 22, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 22, 21 -- X-PMX-Version: 5.1.2.240295 22, 21 -- X-PerlMx-Virus-Scanned: Yes ==============================End of - Headers==============================
Whew... I have been following this thread with great interest. It has reminded me of the times I have heard of falsified data in research. I recall discussing one piece of research with someone else, and this person said "It was justified, due to the seriousness of the subject matter." I just stood there, looking at the person, and blinked my eyes a few times, just to be sure I was awake. I then walked away, shaking my head, and thinking "They are cutting people open, based on bogus research!"
I have recently done an internet search, to see if I could find info on falsified research data, and I did find some very prominent cases. Of the cases I found, I think they all were brought to light by someone bringing it to the attention of the authorities. It is the "Don't poison the well" attitude, and looking the other way, that degrades the entire scientific/medical community, and the hard work of the careful, serious, and caring professionals. I have often said the Scientific Method is dead, when I witness what I refer to as the "New Scientific Method". Decide the outcome of the study, and make the data prove it.
I hope I have not offended anyone, but I also hope that this thread has planted a seed in at least one person who will think twice before looking the other way, or taking the advice "Don't poison the well". I have been told repeatedly that the medical profession is a "Self Policed" one, but have witnessed just the opposite.
These are my opinions, and not those of my employer. I am glad the "surgery" I do is on computer chips only, in an attempt to figure out why they don't work right.
Darrell
pgrover-at-bilbo.bio.purdue.edu wrote on 04/24/2006 01:29:46 PM:
} 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 Randy, } } The question you raised is a good one, and one which is largely ignored } today. It seems we are somehow expected to know this stuff intuitively, or } to absorb it during the mentoring process which (should) constitute graduate } education. I have thought about these things quite a bit during the 12 } years I spent doing construction work following a string of postdocs in } which I witnessed so much scientific misconduct that I lost faith in the } literature. The coup de grace was a micrograph which others claimed to } represent a pure preparation of membrane vesicles, but which turned out (18 } months later when I tracked down the person who took the picture) to be just } a selected field from what was in reality a bunch of heterogeneous glop. I } suddenly realized why I couldn't reproduce the results of the others in the } lab. Then I figured out how two of the others had deceived themselves with } help from a GIGO statistical analysis protocol (random numbers return a } lineweaver-burke R2 of .98!), fear of not getting tenure, and a grad student } who obediently followed his advisor's admonishment "I'm tired of experiments } not working. I want the experiments to start working." Upon consulting, } confidentially, with several senior faculty and administrators, I was } advised "Don't poison the well", and "Just vote with your feet". So I } became a carpenter. BTW, I now do my science as an amateur, and study } things which aren't dependent upon the validity of others' work. (Yes, it's } quite possible to publish from your home address!). } } In a nutshell, everyone's gig is different; nobody can tell you what to do } in a given situation. I adhered to the Yiddish proverb "Tell the truth and } run.", but I was a street-smart kid who had skills to fall back on, and I } have known many who haven't had that luxury. My best advice: KEEP A } METICULOUS LAB NOTEBOOK. My mentor, bless her, demanded that I be most } punctilious in my data recording, something I have found to my dismay is all } too rare today. I would be happy to correspond with you offlist to share my } experiences and thoughts in these matters. } } Paul Grover, Ph.D. } Grover Roofing and Remodeling } } } } } Randy Tindall wrote: } } } ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------------
} } I have a follow-up question to the recent postings on imaging ethics. } } As a technician in a multi-user core facility, what role does someone like } me have in spotting ethically questionable behavior and pointing it out? } Are we obligated to alert a researcher when they may be unconsciously } prejudicing their results by selective imaging? Are we expected to report } it if there is a blatant example of deliberate skewing of results? Is it } okay to let it happen, if we ourselves don't actively participate in it? If } a publication results from a piece of research we assisted with and the } images used to support a conclusion are obviously not representative of } those taken during the research, do we have an obligation to comment? } } I'd be curious to find out how other labs deal with such things. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W125 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 9, 23 -- From TindallR-at-missouri.edu Thu Apr 20 11:12:31 2006 9, 23 -- } Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu } [207.160.151.48]) } 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k3KGCVEp012624 } 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 11:12:31 } -0500 } 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by } um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 9, 23 -- Thu, 20 Apr 2006 11:12:30 -0500 } 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- } Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 } -- Content-Type: text/plain; } 9, 23 -- charset="US-ASCII" } 9, 23 -- Subject: Ethical question } 9, 23 -- Date: Thu, 20 Apr 2006 11:11:39 -0500 9, 23 -- Message-ID: } {BA876152E8653240BE8572E897083EE701849F41-at-UM-EMAIL09.um.umsystem.edu} } 9, 23 -- X-MS-Has-Attach: } 9, 23 -- X-MS-TNEF-Correlator: } 9, 23 -- Thread-Topic: Ethical question } 9, 23 -- thread-index: AcZklRuKX2YC3GZDRjuGFPDySAbWWw== 9, 23 -- From: } "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: } {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 20 Apr 2006 } 16:12:30.0978 (UTC) FILETIME=[39D4A620:01C66495] 9, 23 -- } Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id k3KGCVEp012624 } ==============================End of - Headers============================== } } --------------------------------------------------------------- } In every object there is inexhaustible meaning; the eye sees in } it what the eye brings means of seeing. - Thomas Carlyle } } } } } } ==============================Original Headers============================== } 22, 21 -- From pgrover-at-bilbo.bio.purdue.edu Mon Apr 24 12:28:22 2006 } 22, 21 -- Received: from mailhub128.itcs.purdue.edu (mailhub128. } itcs.purdue.edu [128.210.5.128]) } 22, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k3OHSMX8015525 } 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 Apr 2006 12:28:22 -0500 } 22, 21 -- Received: from paklabpgrover (dhcp155-122.bio.purdue.edu } [128.210.155.122]) } 22, 21 -- by mailhub128.itcs.purdue.edu (8.13.6/8.13.4/internal- } smtp) with ESMTP id k3OHSLCP010642 } 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 Apr 2006 13:28:21 -0400 } 22, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} } 22, 21 -- To: {microscopy-at-microscopy.com} } 22, 21 -- Subject: re: ethical question } 22, 21 -- Date: Mon, 24 Apr 2006 13:28:26 -0400 } 22, 21 -- Message-ID: {000001c667c4$7eb3a070$7a9bd280-at-paklabpgrover} } 22, 21 -- MIME-Version: 1.0 } 22, 21 -- Content-Type: text/plain; } 22, 21 -- charset="us-ascii" } 22, 21 -- Content-Transfer-Encoding: 7bit } 22, 21 -- X-Mailer: Microsoft Office Outlook 11 } 22, 21 -- Thread-Index: AcZnxH5btnNRbsYCThmNsNFgawelDA== } 22, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 22, 21 -- X-PMX-Version: 5.1.2.240295 } 22, 21 -- X-PerlMx-Virus-Scanned: Yes } ==============================End of - Headers==============================
==============================Original Headers============================== 12, 27 -- From milesd-at-us.ibm.com Mon Apr 24 14:34:54 2006 12, 27 -- Received: from e1.ny.us.ibm.com (e1.ny.us.ibm.com [32.97.182.141]) 12, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3OJYrEm027227 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 14:34:53 -0500 12, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 12, 27 -- by e1.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k3OJYqu9008783 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 15:34:52 -0400 12, 27 -- Received: from d01av03.pok.ibm.com (d01av03.pok.ibm.com [9.56.224.217]) 12, 27 -- by d01relay02.pok.ibm.com (8.12.10/NCO/VER6.8) with ESMTP id k3OJYq6U236336 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 15:34:52 -0400 12, 27 -- Received: from d01av03.pok.ibm.com (loopback [127.0.0.1]) 12, 27 -- by d01av03.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k3OJYqmA008133 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 15:34:52 -0400 12, 27 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 12, 27 -- by d01av03.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k3OJYqDg008122 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 15:34:52 -0400 12, 27 -- In-Reply-To: {200604241729.k3OHTkrA016944-at-ns.microscopy.com} 12, 27 -- Subject: Re: [Microscopy] re: ethical question 12, 27 -- To: Microscopy-at-MSA.Microscopy.Com 12, 27 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 12, 27 -- Message-ID: {OF65CB53D7.88540C3F-ON8525715A.00686DEA-8525715A.006B8F6E-at-us.ibm.com} 12, 27 -- From: Darrell Miles {milesd-at-us.ibm.com} 12, 27 -- Date: Mon, 24 Apr 2006 15:34:51 -0400 12, 27 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 6.5.4FP3 HF3|February 22, 2006) at 12, 27 -- 04/24/2006 15:34:52 12, 27 -- MIME-Version: 1.0 12, 27 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Just for the record (I didn't make it very clear before), neither I, nor the coworker, had anything to do with the research we were discussing which left me shaking my head at his statement. It was not even in our field (electronics), but I feel the question of ethics pervades all of science.
Darrell Miles/Fishkill/IBM-at-IBMUS wrote on 04/24/2006 03:35:49 PM:
} 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 } ----------------------------------------------------------------------------
} } Whew... I have been following this thread with great interest. It has } reminded me of the times I have heard of falsified data in research. I } recall discussing one piece of research with someone else, and this person } said "It was justified, due to the seriousness of the subject matter." I } just stood there, looking at the person, and blinked my eyes a few times, } just to be sure I was awake. I then walked away, shaking my head, and } thinking "They are cutting people open, based on bogus research!" } } I have recently done an internet search, to see if I could find info on } falsified research data, and I did find some very prominent cases. Of the } cases I found, I think they all were brought to light by someone bringing } it to the attention of the authorities. It is the "Don't poison the well" } attitude, and looking the other way, that degrades the entire } scientific/medical community, and the hard work of the careful, serious, } and caring professionals. I have often said the Scientific Method is dead, } when I witness what I refer to as the "New Scientific Method". Decide the } outcome of the study, and make the data prove it. } } I hope I have not offended anyone, but I also hope that this thread has } planted a seed in at least one person who will think twice before looking } the other way, or taking the advice "Don't poison the well". I have been } told repeatedly that the medical profession is a "Self Policed" one, but } have witnessed just the opposite. } } These are my opinions, and not those of my employer. I am glad the } "surgery" I do is on computer chips only, in an attempt to figure out why } they don't work right. } } Darrell } } pgrover-at-bilbo.bio.purdue.edu wrote on 04/24/2006 01:29:46 PM: } } } } } } } } } } ----------------------------------------------------------------------------
} } } 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 Randy, } } } } The question you raised is a good one, and one which is largely ignored } } today. It seems we are somehow expected to know this stuff intuitively, } or } } to absorb it during the mentoring process which (should) constitute } graduate } } education. I have thought about these things quite a bit during the 12 } } years I spent doing construction work following a string of postdocs in } } which I witnessed so much scientific misconduct that I lost faith in the } } literature. The coup de grace was a micrograph which others claimed to } } represent a pure preparation of membrane vesicles, but which turned out } (18 } } months later when I tracked down the person who took the picture) to be } just } } a selected field from what was in reality a bunch of heterogeneous glop. } I } } suddenly realized why I couldn't reproduce the results of the others in } the } } lab. Then I figured out how two of the others had deceived themselves } with } } help from a GIGO statistical analysis protocol (random numbers return a } } lineweaver-burke R2 of .98!), fear of not getting tenure, and a grad } student } } who obediently followed his advisor's admonishment "I'm tired of } experiments } } not working. I want the experiments to start working." Upon consulting, } } confidentially, with several senior faculty and administrators, I was } } advised "Don't poison the well", and "Just vote with your feet". So I } } became a carpenter. BTW, I now do my science as an amateur, and study } } things which aren't dependent upon the validity of others' work. (Yes, } it's } } quite possible to publish from your home address!). } } } } In a nutshell, everyone's gig is different; nobody can tell you what to } do } } in a given situation. I adhered to the Yiddish proverb "Tell the truth } and } } run.", but I was a street-smart kid who had skills to fall back on, and I } } have known many who haven't had that luxury. My best advice: KEEP A } } METICULOUS LAB NOTEBOOK. My mentor, bless her, demanded that I be most } } punctilious in my data recording, something I have found to my dismay is } all } } too rare today. I would be happy to correspond with you offlist to share } my } } experiences and thoughts in these matters. } } } } Paul Grover, Ph.D. } } Grover Roofing and Remodeling } } } } } } } } } } Randy Tindall wrote: } } } } } } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------------
} } } } } I have a follow-up question to the recent postings on imaging ethics. } } } } As a technician in a multi-user core facility, what role does someone } like } } me have in spotting ethically questionable behavior and pointing it out? } } Are we obligated to alert a researcher when they may be unconsciously } } prejudicing their results by selective imaging? Are we expected to } report } } it if there is a blatant example of deliberate skewing of results? Is it } } okay to let it happen, if we ourselves don't actively participate in it? } If } } a publication results from a piece of research we assisted with and the } } images used to support a conclusion are obviously not representative of } } those taken during the research, do we have an obligation to comment? } } } } I'd be curious to find out how other labs deal with such things. } } } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility---We Do Small Well! } } W125 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-2227 } } Email: tindallr-at-missouri.edu } } Web: http://www.emc.missouri.edu } } } } } } } } } } } } } } } } ==============================Original } Headers============================== } } 9, 23 -- From TindallR-at-missouri.edu Thu Apr 20 11:12:31 2006 9, 23 -- } } Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu } } [207.160.151.48]) } } 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } k3KGCVEp012624 } } 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Apr 2006 11:12:31 } } -0500 } } 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by } } um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } } 9, 23 -- Thu, 20 Apr 2006 11:12:30 -0500 } } 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- } } Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, } 23 } } -- Content-Type: text/plain; } } 9, 23 -- charset="US-ASCII" } } 9, 23 -- Subject: Ethical question } } 9, 23 -- Date: Thu, 20 Apr 2006 11:11:39 -0500 9, 23 -- Message-ID: } } {BA876152E8653240BE8572E897083EE701849F41-at-UM-EMAIL09.um.umsystem.edu} } } 9, 23 -- X-MS-Has-Attach: } } 9, 23 -- X-MS-TNEF-Correlator: } } 9, 23 -- Thread-Topic: Ethical question } } 9, 23 -- thread-index: AcZklRuKX2YC3GZDRjuGFPDySAbWWw== 9, 23 -- From: } } "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: } } {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 20 Apr 2006 } } 16:12:30.0978 (UTC) FILETIME=[39D4A620:01C66495] 9, 23 -- } } Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from } } quoted-printable to 8bit by ns.microscopy.com id k3KGCVEp012624 } } ==============================End of - } Headers============================== } } } } --------------------------------------------------------------- } } In every object there is inexhaustible meaning; the eye sees in } } it what the eye brings means of seeing. - Thomas Carlyle } } } } } } } } } } } } ==============================Original } Headers============================== } } 22, 21 -- From pgrover-at-bilbo.bio.purdue.edu Mon Apr 24 12:28:22 2006 } } 22, 21 -- Received: from mailhub128.itcs.purdue.edu (mailhub128. } } itcs.purdue.edu [128.210.5.128]) } } 22, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k3OHSMX8015525 } } 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 Apr 2006 12:28:22 } -0500 } } 22, 21 -- Received: from paklabpgrover (dhcp155-122.bio.purdue.edu } } [128.210.155.122]) } } 22, 21 -- by mailhub128.itcs.purdue.edu (8.13.6/8.13.4/internal- } } smtp) with ESMTP id k3OHSLCP010642 } } 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 Apr 2006 13:28:21 } -0400 } } 22, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} } } 22, 21 -- To: {microscopy-at-microscopy.com} } } 22, 21 -- Subject: re: ethical question } } 22, 21 -- Date: Mon, 24 Apr 2006 13:28:26 -0400 } } 22, 21 -- Message-ID: {000001c667c4$7eb3a070$7a9bd280-at-paklabpgrover} } } 22, 21 -- MIME-Version: 1.0 } } 22, 21 -- Content-Type: text/plain; } } 22, 21 -- charset="us-ascii" } } 22, 21 -- Content-Transfer-Encoding: 7bit } } 22, 21 -- X-Mailer: Microsoft Office Outlook 11 } } 22, 21 -- Thread-Index: AcZnxH5btnNRbsYCThmNsNFgawelDA== } } 22, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } } 22, 21 -- X-PMX-Version: 5.1.2.240295 } } 22, 21 -- X-PerlMx-Virus-Scanned: Yes } } ==============================End of - } Headers============================== } } } ==============================Original Headers============================== } 12, 27 -- From milesd-at-us.ibm.com Mon Apr 24 14:34:54 2006 } 12, 27 -- Received: from e1.ny.us.ibm.com (e1.ny.us.ibm.com [32.97.182.141]) } 12, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k3OJYrEm027227 } 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 } 14:34:53 -0500 } 12, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm. } com [9.56.227.234]) } 12, 27 -- by e1.ny.us.ibm.com (8.12.11.20060308/8.12.11) with } ESMTP id k3OJYqu9008783 } 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 } 15:34:52 -0400 } 12, 27 -- Received: from d01av03.pok.ibm.com (d01av03.pok.ibm.com } [9.56.224.217]) } 12, 27 -- by d01relay02.pok.ibm.com (8.12.10/NCO/VER6.8) with } ESMTP id k3OJYq6U236336 } 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 } 15:34:52 -0400 } 12, 27 -- Received: from d01av03.pok.ibm.com (loopback [127.0.0.1]) } 12, 27 -- by d01av03.pok.ibm.com (8.12.11/8.13.3) with ESMTP id } k3OJYqmA008133 } 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 } 15:34:52 -0400 } 12, 27 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com } [9.56.229.50]) } 12, 27 -- by d01av03.pok.ibm.com (8.12.11/8.12.11) with ESMTP id } k3OJYqDg008122 } 12, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 } 15:34:52 -0400 } 12, 27 -- In-Reply-To: {200604241729.k3OHTkrA016944-at-ns.microscopy.com} } 12, 27 -- Subject: Re: [Microscopy] re: ethical question } 12, 27 -- To: Microscopy-at-MSA.Microscopy.Com } 12, 27 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 } 12, 27 -- Message-ID: {OF65CB53D7.88540C3F-ON8525715A. } 00686DEA-8525715A.006B8F6E-at-us.ibm.com} } 12, 27 -- From: Darrell Miles {milesd-at-us.ibm.com} } 12, 27 -- Date: Mon, 24 Apr 2006 15:34:51 -0400 } 12, 27 -- X-MIMETrack: Serialize by Router on } D01ML076/01/M/IBM(Release 6.5.4FP3 HF3|February 22, 2006) at } 12, 27 -- 04/24/2006 15:34:52 } 12, 27 -- MIME-Version: 1.0 } 12, 27 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - Headers==============================
==============================Original Headers============================== 10, 27 -- From milesd-at-us.ibm.com Mon Apr 24 18:19:45 2006 10, 27 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3ONJj0t008172 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 18:19:45 -0500 10, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 10, 27 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k3ONJjhU018652 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 19:19:45 -0400 10, 27 -- Received: from d01av03.pok.ibm.com (d01av03.pok.ibm.com [9.56.224.217]) 10, 27 -- by d01relay02.pok.ibm.com (8.12.10/NCO/VER6.8) with ESMTP id k3ONJi6U232030 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 19:19:45 -0400 10, 27 -- Received: from d01av03.pok.ibm.com (loopback [127.0.0.1]) 10, 27 -- by d01av03.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k3ONJi48016305 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 19:19:44 -0400 10, 27 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 10, 27 -- by d01av03.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k3ONJiXA016293 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 24 Apr 2006 19:19:44 -0400 10, 27 -- In-Reply-To: {200604241935.k3OJZnGo028639-at-ns.microscopy.com} 10, 27 -- Subject: Re: [Microscopy] ethical question 10, 27 -- To: Microscopy-at-MSA.Microscopy.Com 10, 27 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 10, 27 -- Message-ID: {OF2869E5C1.50BC4ACC-ON8525715A.007F7279-8525715A.008025AE-at-us.ibm.com} 10, 27 -- From: Darrell Miles {milesd-at-us.ibm.com} 10, 27 -- Date: Mon, 24 Apr 2006 19:19:42 -0400 10, 27 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 6.5.4FP3 HF3|February 22, 2006) at 10, 27 -- 04/24/2006 19:19:44 10, 27 -- MIME-Version: 1.0 10, 27 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Paul I could not have said better. And the advisor's admonishment I heard too during my PhD thesis. Bad recall. And cheers to the notebook.
Stéphane
--- pgrover-at-bilbo.bio.purdue.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi Randy, } } The question you raised is a good one, and one which } is largely ignored } today. It seems we are somehow expected to know } this stuff intuitively, or } to absorb it during the mentoring process which } (should) constitute graduate } education. I have thought about these things quite } a bit during the 12 } years I spent doing construction work following a } string of postdocs in } which I witnessed so much scientific misconduct that } I lost faith in the } literature. The coup de grace was a micrograph } which others claimed to } represent a pure preparation of membrane vesicles, } but which turned out (18 } months later when I tracked down the person who took } the picture) to be just } a selected field from what was in reality a bunch of } heterogeneous glop. I } suddenly realized why I couldn't reproduce the } results of the others in the } lab. Then I figured out how two of the others had } deceived themselves with } help from a GIGO statistical analysis protocol } (random numbers return a } lineweaver-burke R2 of .98!), fear of not getting } tenure, and a grad student } who obediently followed his advisor's admonishment } "I'm tired of experiments } not working. I want the experiments to start } working." Upon consulting, } confidentially, with several senior faculty and } administrators, I was } advised "Don't poison the well", and "Just vote with } your feet". So I } became a carpenter. BTW, I now do my science as an } amateur, and study } things which aren't dependent upon the validity of } others' work. (Yes, it's } quite possible to publish from your home address!). } } In a nutshell, everyone's gig is different; nobody } can tell you what to do } in a given situation. I adhered to the Yiddish } proverb "Tell the truth and } run.", but I was a street-smart kid who had skills } to fall back on, and I } have known many who haven't had that luxury. My } best advice: KEEP A } METICULOUS LAB NOTEBOOK. My mentor, bless her, } demanded that I be most } punctilious in my data recording, something I have } found to my dismay is all } too rare today. I would be happy to correspond with } you offlist to share my } experiences and thoughts in these matters. } } Paul Grover, Ph.D. } Grover Roofing and Remodeling } } } } } Randy Tindall wrote: } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a follow-up question to the recent postings } on imaging ethics. } } As a technician in a multi-user core facility, what } role does someone like } me have in spotting ethically questionable behavior } and pointing it out? } Are we obligated to alert a researcher when they may } be unconsciously } prejudicing their results by selective imaging? Are } we expected to report } it if there is a blatant example of deliberate } skewing of results? Is it } okay to let it happen, if we ourselves don't } actively participate in it? If } a publication results from a piece of research we } assisted with and the } images used to support a conclusion are obviously } not representative of } those taken during the research, do we have an } obligation to comment? } } I'd be curious to find out how other labs deal with } such things. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small } Well! } W125 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original } Headers============================== } 9, 23 -- From TindallR-at-missouri.edu Thu Apr 20 } 11:12:31 2006 9, 23 -- } Received: from um-exproto8.um.umsystem.edu } (um-exproto8.um.umsystem.edu } [207.160.151.48]) } 9, 23 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k3KGCVEp012624 } 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 } Apr 2006 11:12:31 } -0500 } 9, 23 -- Received: from UM-EMAIL09.um.umsystem.edu } ([207.160.151.31]) by } um-exproto8.um.umsystem.edu with Microsoft } SMTPSVC(6.0.3790.1830); } 9, 23 -- Thu, 20 Apr 2006 11:12:30 -0500 } 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange } V6.5 9, 23 -- } Content-class: urn:content-classes:message 9, 23 -- } MIME-Version: 1.0 9, 23 } -- Content-Type: text/plain; } 9, 23 -- charset="US-ASCII" } 9, 23 -- Subject: Ethical question } 9, 23 -- Date: Thu, 20 Apr 2006 11:11:39 -0500 9, 23 } -- Message-ID: } {BA876152E8653240BE8572E897083EE701849F41-at-UM-EMAIL09.um.umsystem.edu} } 9, 23 -- X-MS-Has-Attach: } 9, 23 -- X-MS-TNEF-Correlator: } 9, 23 -- Thread-Topic: Ethical question } 9, 23 -- thread-index: } AcZklRuKX2YC3GZDRjuGFPDySAbWWw== 9, 23 -- From: } "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- } To: } {microscopy-at-microscopy.com} 9, 23 -- } X-OriginalArrivalTime: 20 Apr 2006 } 16:12:30.0978 (UTC) FILETIME=[39D4A620:01C66495] 9, } 23 -- } Content-Transfer-Encoding: 8bit 9, 23 -- } X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } k3KGCVEp012624 } ==============================End of - } Headers============================== } } --------------------------------------------------------------- } In every object there is inexhaustible meaning; the } eye sees in } it what the eye brings means of seeing. - Thomas } Carlyle } } } } } } ==============================Original } Headers============================== } 22, 21 -- From pgrover-at-bilbo.bio.purdue.edu Mon Apr } 24 12:28:22 2006 } 22, 21 -- Received: from mailhub128.itcs.purdue.edu } (mailhub128.itcs.purdue.edu [128.210.5.128]) } 22, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k3OHSMX8015525 } 22, 21 -- for {microscopy-at-microscopy.com} ; Mon, 24 } Apr === message truncated ===
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Hi and sorry if this question does not really enter the field of microscopy but this list being full of clever helpful people I would be fool not to rely on it.
I cannot explain my subject of research in detail, so I will not give many informations. I would just ask you not to propose me other ways to do the experiment, I have to bear with this way for any reason.
I fix BSA with glutaraldehyde in 10 mM phosphate buffer pH 7.2 and I need to neutralize the glutaraldehyde in the solution. I need a way to keep everything in solution while neutralizing the fixation process (aldehyde+amine groups). I cannot move the BSA from the solution, but I could remove the glutar (although I wonder how). I was thinking about increasing the pH. Do you think glutar loses its fixating activity at pH 10? Any idea?
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
We sometimes have concerns about the cross-linking activity of glutaraldehyde when doing preparations of virus suspensions where we do not want artificial aggregation. Simply put, dogma states that because glutaraldehyde is a 5 carbon chain with highly reactive carboxyl groups at each end it is more likely to cross link different virions, or virions with cellular detritis, than formaldehyde, which has a single carbon and a single reactive aldehyde group. Normally I use glutaraldehyde at a concentration of 0.1%I to stabilize reactions. Over the years I have not really seen appreciable clumping which could beassociated with the fizative. But if you are doing an immunoprecipitation style IEM procedure you really don't want to take the chance of creating an artificial situation. To protect against this I neutralize the fixatives in the sample by addition of glycine. Note: lysine is frequently used. However, it has two reactive amine sites, so I avoid that because, technically, it may also contribute to cross linking. The final concentration of glycine we use in the preparation is 8mM to neutralize 0.1% Glutaraldehyde, and 125mM to neutralize 2% Paraformaldehyde. Perhaps a Chemistry minded member of the list will be able to provide for a better way of neutralizing.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit Medical Microbiology and Infectious Diseases University of Manitoba 531 Basic Medical Sciences 730 William Avenue Winnipeg, Manitoba, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Telephone: 204-789-3313 Fax: 204-789-3926 Pager 204-931-9354 Cell 204-781-1502 Home Telephone: 204-489-6924
==============================Original Headers============================== 7, 21 -- From paul_hazelton-at-umanitoba.ca Tue Apr 25 09:26:38 2006 7, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3PEQbsP026240 7, 21 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 09:26:38 -0500 7, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 7, 21 -- (authenticated bits=0) 7, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k3PEQadT006840; 7, 21 -- Tue, 25 Apr 2006 09:26:36 -0500 (CDT) 7, 21 -- Message-ID: {444E3197.9000804-at-umanitoba.ca} 7, 21 -- Date: Tue, 25 Apr 2006 09:26:31 -0500 7, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 7, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 7, 21 -- X-Accept-Language: en-us, en 7, 21 -- MIME-Version: 1.0 7, 21 -- To: nizets2-at-yahoo.com 7, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 7, 21 -- Subject: Re: [Microscopy] neutralizing glutaraldehyde 7, 21 -- References: {200604250846.k3P8kXnL006763-at-ns.microscopy.com} 7, 21 -- In-Reply-To: {200604250846.k3P8kXnL006763-at-ns.microscopy.com} 7, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
perhaps an alternative would be NH4Cl (ammonium chloride), applied in a washing buffer, end concentration 50mM, application time: necessarily only seconds (personal comment by Prof. Roth) , but usually for tissue specimens (as used in TEM-specimen preparations) 20-30 [up to 4 hrs] min, followed by one to two additional washes in the respective, pure buffer solution [e.g. 4 degr.C, over night]. Source: ROTH J. et al. : Enhancement of Structural Preservation and (EPON) LOWICRYL K4M Low Temp. Immunocytochemical Staining in Low Temperature Embedded Pancreatic Tissue. J.Histochem.Cytochem. 29, 663-671,1981
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Stephane
We sometimes have concerns about the cross-linking activity of glutaraldehyde when doing preparations of virus suspensions where we do not want artificial aggregation. Simply put, dogma states that because glutaraldehyde is a 5 carbon chain with highly reactive carboxyl groups at each end it is more likely to cross link different virions, or virions with cellular detritis, than formaldehyde, which has a single carbon and a single reactive aldehyde group. Normally I use glutaraldehyde at a concentration of 0.1%I to stabilize reactions. Over the years I have not really seen appreciable clumping which could be associated with the fixative. But if you are doing an immunoprecipitation style IEM procedure you really don't want to take the chance of creating an artificial situation. To protect against this I neutralize the fixatives in the sample by addition of glycine. Note: lysine is frequently used. However, it has two reactive amine sites, so I avoid that because, technically, it may also contribute to cross linking. The final concentration of glycine we use in the preparation is 8mM to neutralize 0.1% Glutaraldehyde, and 125mM to neutralize 2% Paraformaldehyde. Perhaps a Chemistry minded member of the list will be able to provide for a better way of neutralizing.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit Medical Microbiology and Infectious Diseases University of Manitoba 531 Basic Medical Sciences 730 William Avenue Winnipeg, Manitoba, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Telephone: 204-789-3313 Fax: 204-789-3926 Pager 204-931-9354 Cell 204-781-1502 Home Telephone: 204-489-6924
==============================Original Headers============================== 7, 21 -- From paul_hazelton-at-umanitoba.ca Tue Apr 25 09:26:38 2006 7, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3PEQbsP026240 7, 21 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 09:26:38 -0500 7, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 7, 21 -- (authenticated bits=0) 7, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k3PEQadT006840; 7, 21 -- Tue, 25 Apr 2006 09:26:36 -0500 (CDT) 7, 21 -- Message-ID: {444E3197.9000804-at-umanitoba.ca} 7, 21 -- Date: Tue, 25 Apr 2006 09:26:31 -0500 7, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 7, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 7, 21 -- X-Accept-Language: en-us, en 7, 21 -- MIME-Version: 1.0 7, 21 -- To: nizets2-at-yahoo.com 7, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 7, 21 -- Subject: Re: [Microscopy] neutralizing glutaraldehyde 7, 21 -- References: {200604250846.k3P8kXnL006763-at-ns.microscopy.com} 7, 21 -- In-Reply-To: {200604250846.k3P8kXnL006763-at-ns.microscopy.com} 7, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 18, 28 -- From W.Muss-at-salk.at Tue Apr 25 09:48:43 2006 18, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 18, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3PEmgu5004059 18, 28 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 09:48:43 -0500 18, 28 -- Received: from localhost (localhost [127.0.0.1]) 18, 28 -- by hermes.lks.at (Postfix) with ESMTP id 72A415A900A; 18, 28 -- Tue, 25 Apr 2006 16:48:40 +0200 (CEST) 18, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 18, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 18, 28 -- with ESMTP id 83185-07; Tue, 25 Apr 2006 16:48:40 +0200 (CEST) 18, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 18, 28 -- by hermes.lks.at (Postfix) with SMTP id 092ED5A9020; 18, 28 -- Tue, 25 Apr 2006 16:48:40 +0200 (CEST) 18, 28 -- Received: by localhost with Microsoft MAPI; Tue, 25 Apr 2006 16:48:39 +0200 18, 28 -- Message-ID: {01C66888.1A7D5460.W.Muss-at-salk.at} 18, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 18, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 18, 28 -- To: "'paul_hazelton-at-umanitoba.ca'" {paul_hazelton-at-umanitoba.ca} 18, 28 -- Cc: "'microscopy-at-microscopy.com'" {microscopy-at-microscopy.com} 18, 28 -- Subject: [Microscopy] Re: neutralizing glutaraldehyde 18, 28 -- Date: Tue, 25 Apr 2006 16:48:38 +0200 18, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 18, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 18, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 18, 28 -- MIME-Version: 1.0 18, 28 -- Content-Type: text/plain; charset="us-ascii" 18, 28 -- Content-Transfer-Encoding: 7bit 18, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Hello, I came across a reference where they used a 1% saturated solution of thiocarbohydrazide (TCH) after fixation with Glutaraldehyde and Osmium tetroxide. They found a better preservation of bacterial-root structures than standard fixation alone. Has anyone tried this technique or variants of it? Any recommendations? Thanks
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 2, 24 -- From gvrdolja-at-nature.berkeley.edu Tue Apr 25 10:21:36 2006 2, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 2, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3PFLafJ014775 2, 24 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 10:21:36 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 67B4CC1E71 2, 24 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 08:21:36 -0700 (PDT) 2, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 2, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 2, 24 -- with ESMTP id 27405-02-2 for {microscopy-at-microscopy.com} ; 2, 24 -- Tue, 25 Apr 2006 08:21:23 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id 3456EC1E77; Tue, 25 Apr 2006 08:21:23 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 2A643C1E71 2, 24 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 08:21:23 -0700 (PDT) 2, 24 -- Date: Tue, 25 Apr 2006 08:21:22 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: microscopy-at-microscopy.com 2, 24 -- Subject: thiocarbohydrazide solution 2, 24 -- Message-ID: {Pine.SOC.4.64.0604250819080.25182-at-nature.Berkeley.EDU} 2, 24 -- MIME-Version: 1.0 2, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 2, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
Would it be possible to add a concentrated solution of tris (or even dissolve crystaline tris into the BSA solution) to bind any remaining reactive glutaraldehyde. This would raise the pH though. One could also use a concentrated solution of glycine or even lysine if one wanted an additional amino group to aid in the binding. If this concentrated amino acid solution were pHed before addition to your BSA experiment I don't think it would alter the pH much.
Good luck, George
George P. Leser, PhD Dept. Biochemistry, Molecular Biology, and Cell Biology Northwestern University Hogan 2-100 2153 North Campus Drive Evanston, IL 60208
g-leser-at-northwestern.edu
----- Original Message ----- X-from: {nizets2-at-yahoo.com} To: {g-leser-at-northwestern.edu} Sent: Tuesday, April 25, 2006 3:48 AM
HI, Gordon- We use it all of the time for cultured cells. I can come up with a protocol for you within a day or two. Carol
} -------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgsu.edu http://www.bgsu.edu/departments/biology/facilities/MnM ___________________________________________________________________________
Thanks to all those of you who have replied to my post. I had no idea there were so many of us. I promise to reply to all of you as I get time. Meanwhile I think it would be wonderful if everyone else could see the similarities in the self-deception we've all witnessed. And I certainly don't claim to be immune; I'm sure I deceive myself also. We're all fools to some extent, but where does it cross the fool-to-knave boundary?
It's certainly a topic worthy of discussion, but probably has used enough bandwidth here. If anyone is interested in some sort of forum, I'd love it. A great weight has been lifted from me just by your posts today. The problem is, I'm not so technology oriented that I would know how to set up something like that. But as I said, I have been mulling these things over for many years, and I have SPECIFIC IDEAS about what can be done to IMPROVE OUR PROFESSION, heal the wounds and wound the heels. I don't just want to commiserate. I want to light a candle rather than curse the darkness, and all that.
I have a Thoreau quote on my bookshelf: "In what concerns you much, do not think that you have companions. Know that you are alone in the world." I'm taking it down. :o)
Paul
--------------------------------------------------------------- In every object there is inexhaustible meaning; the eye sees in it what the eye brings means of seeing. - Thomas Carlyle
==============================Original Headers============================== 9, 21 -- From pgrover-at-bilbo.bio.purdue.edu Tue Apr 25 16:26:59 2006 9, 21 -- Received: from mailhub131.itcs.purdue.edu (mailhub131.itcs.purdue.edu [128.210.5.131]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3PLQx9w018523 9, 21 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 16:26:59 -0500 9, 21 -- Received: from paklabpgrover (dhcp155-122.bio.purdue.edu [128.210.155.122]) 9, 21 -- by mailhub131.itcs.purdue.edu (8.13.6/8.13.4/internal-smtp) with ESMTP id k3PLQw2c017588 9, 21 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 17:26:59 -0400 9, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} 9, 21 -- To: {microscopy-at-microscopy.com} 9, 21 -- Subject: re: ethical question 9, 21 -- Date: Tue, 25 Apr 2006 17:26:59 -0400 9, 21 -- Message-ID: {000001c668ae$fcd036e0$7a9bd280-at-paklabpgrover} 9, 21 -- MIME-Version: 1.0 9, 21 -- Content-Type: text/plain; 9, 21 -- charset="us-ascii" 9, 21 -- Content-Transfer-Encoding: 7bit 9, 21 -- X-Mailer: Microsoft Office Outlook 11 9, 21 -- Thread-Index: AcZorvyGo6QTb8S4Tf+nKRODZefCeA== 9, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 9, 21 -- X-PMX-Version: 5.1.2.240295 9, 21 -- X-PerlMx-Virus-Scanned: Yes ==============================End of - Headers==============================
Dear List, For a TEM with a FEG there are several parameters that can have an effect on the coherence of the beam: the gun lens setting, the extraction voltage, the spot size setting, and the C2 aperture size. For a given beam current and diameter at the specimen, which of the settings for these parameters will produce the most coherent beam; i.e., will using a smaller C2 aperture and a smaller spot size number be better than using a larger C2 aperture and spot size number, or will coherence be better with smaller C2 aperture, larger spot size number, and higher extraction voltage, etc.? I have tried to find this out in electron optics books with no luck so far, so if anyone can point me to the appropriate books or articles, I'd be grateful. TIA. 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============================== 2, 20 -- From tivol-at-caltech.edu Tue Apr 25 16:51:55 2006 2, 20 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3PLptOM028658 2, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Apr 2006 16:51:55 -0500 2, 20 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 2, 20 -- by water-ox-postvirus (Postfix) with ESMTP id 9A3D034520 2, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Apr 2006 14:51:54 -0700 (PDT) 2, 20 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 2, 20 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id B3D4935FCB 2, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Apr 2006 14:51:53 -0700 (PDT) 2, 20 -- Mime-Version: 1.0 (Apple Message framework v623) 2, 20 -- Content-Transfer-Encoding: 7bit 2, 20 -- Message-Id: {6f4fd52b762726fbbbb2f81788586736-at-caltech.edu} 2, 20 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 2, 20 -- To: microscopy-at-msa.microscopy.com 2, 20 -- From: Bill Tivol {tivol-at-caltech.edu} 2, 20 -- Subject: Coherence in a TEM 2, 20 -- Date: Tue, 25 Apr 2006 15:01:56 -0700 2, 20 -- X-Mailer: Apple Mail (2.623) 2, 20 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both dyel-at-mail.nih.gov as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dyel-at-mail.nih.gov Name: Chip Dye
Organization: NIH
Title-Subject: [Filtered] Gelatin
Question: Dear ListServers,
Does anyone have experience cutting fixed tissue/insect brains using gelatin as the embedding media? What should the strength (bloom#) be? What percentage? I plan on using my ultramicrotome and a glass knife (at ambient temps.) to make about 60-80 micron sections. I will eventually apply ICC to these sections.
Any advice or suggestions are greatly appreciated!
I use a vibratome instead of the ultramicrotome for this. The insect brains are fixed and then embedded in warm (40°) 15% gelatin (any type of gelatin should be useful). When the gelatin has set in the fridge, I cut a little block out of it and section it on the vibratome. 50microns for light microscopy, 70microns for pre-embedding TEM immunocytochemistry.
Gerd
} Question: Dear ListServers, } } Does anyone have experience cutting fixed tissue/insect brains using gelatin as the embedding media? What should the strength (bloom#) be? What percentage? I plan on using my ultramicrotome and a glass knife (at ambient temps.) to make about 60-80 micron sections. I will eventually apply ICC to these sections. } } Any advice or suggestions are greatly appreciated! } } Thank you, } } Chip Dye } } } Microscopist } } Microscopy & Imaging Core, NICHD, NIH http://mic.nichd.nih.gov } Building 49, Room 5W-14 } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } Phone: 301-496-3627 } E-mail: dyel-at-mail.nih.gov } } } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 15, 12 -- From zaluzec-at-microscopy.com Tue Apr 25 20:24:27 2006 } 15, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 15, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3Q1OOkp008816 } 15, 12 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 20:24:26 -0500 } 15, 12 -- Mime-Version: 1.0 } 15, 12 -- X-Sender: (Unverified) } 15, 12 -- Message-Id: {p06110402c0747c2b6167-at-[206.69.208.22]} } 15, 12 -- Date: Tue, 25 Apr 2006 20:24:23 -0500 } 15, 12 -- To: microscopy-at-microscopy.com } 15, 12 -- From: dyel-at-mail.nih.gov (by way of MicroscopyListserver) } 15, 12 -- Subject: viaWWW: Gelatin as the embedding media } 15, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
-- Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Zentrum für Molekulare Medizin Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
Besides museums;-) is there a market for darkroom enlargers? I've tried Adorama and KEH. I'd appreciate any leads. Best, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu] Sent: Wednesday, April 26, 2006 9:37 AM To: Mike Bode
Besides museums;-) is there a market for darkroom enlargers? I've tried Adorama and KEH. I'd appreciate any leads. Best, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
On Wed Apr 26 03:46:47 EDT 2006, 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 } ---------------------------------------------------------------------------- } } Dear Chip Dye, } } I use a vibratome instead of the ultramicrotome for this. } The insect brains are fixed and then embedded in warm (40?) 15% } gelatin (any type of gelatin should be useful). When the gelatin } has set in the fridge, I cut a little block out of it and section } it on the vibratome. } 50microns for light microscopy, 70microns for pre-embedding TEM } immunocytochemistry. } } Gerd } } Hello, For animal tissue I prefer to use agar embedding: it holds sections better when processed for example for further flat embedding of ICC. I also cut sections with vibratome, yet I use sapphire knife instead of the standard blade to get sections about 35-40 micron thick which I prefer for ICC. Hope this helps, Albina
} } Question: Dear ListServers, } } } } Does anyone have experience cutting fixed tissue/insect brains } } using gelatin as the embedding media? What should the strength } } (bloom#) be? What percentage? I plan on using my } } ultramicrotome and a glass knife (at ambient temps.) to make } } about 60-80 micron sections. I will eventually apply ICC to } } these sections. } } } } Any advice or suggestions are greatly appreciated! } } } } Thank you, } } } } Chip Dye } } } } } } Microscopist } } } } Microscopy & Imaging Core, NICHD, NIH http://mic.nichd.nih.gov } } Building 49, Room 5W-14 } } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } } Phone: 301-496-3627 E-mail: dyel-at-mail.nih.gov } } } } } } } } --------------------------------------------------------------------------- } } ==============================Original } } Headers============================== } } 15, 12 -- From zaluzec-at-microscopy.com Tue Apr 25 20:24:27 2006 } } 15, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com } } [206.69.208.22]) } } 15, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k3Q1OOkp008816 } } 15, 12 -- for {microscopy-at-microscopy.com} ; Tue, 25 Apr 2006 } } 20:24:26 -0500 } } 15, 12 -- Mime-Version: 1.0 } } 15, 12 -- X-Sender: (Unverified) } } 15, 12 -- Message-Id: {p06110402c0747c2b6167-at-[206.69.208.22]} } } 15, 12 -- Date: Tue, 25 Apr 2006 20:24:23 -0500 } } 15, 12 -- To: microscopy-at-microscopy.com } } 15, 12 -- From: dyel-at-mail.nih.gov (by way of } } MicroscopyListserver) } } 15, 12 -- Subject: viaWWW: Gelatin as the embedding media } } 15, 12 -- Content-Type: text/plain; charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } -- Dr. Gerd Leitinger } } Institut f?r Zellbiologie, Histologie und Embryologie } Zentrum f?r Molekulare Medizin } Medizinische Universit?t Graz } Harrachgasse 21 } A-8010 Graz } Austria } } Tel. ++43 316 380 4237 } Fax. ++43 316 380 9625 } Mailto: Gerd.Leitinger-at-meduni-graz.at } } } ==============================Original } Headers============================== } 11, 20 -- From gerd.leitinger-at-meduni-graz.at Wed Apr 26 02:40:48 } 2006 } 11, 20 -- Received: from herakles.kfunigraz.ac.at } (HERAKLES.kfunigraz.ac.at [143.50.13.36]) } 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k3Q7el0C023530 } 11, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 26 Apr 2006 } 02:40:48 -0500 } 11, 20 -- Received: from [10.200.112.73] ([193.170.104.112]) by } herakles.kfunigraz.ac.at with Microsoft SMTPSVC(5.0.2195.6713); } 11, 20 -- Wed, 26 Apr 2006 09:40:47 +0200 } 11, 20 -- From: "Gerd Leitinger" {gerd.leitinger-at-meduni-graz.at} } 11, 20 -- To: dyel-at-mail.nih.gov, Microscopy-at-microscopy.com } 11, 20 -- Date: Wed, 26 Apr 2006 09:40:30 +0200 } 11, 20 -- MIME-Version: 1.0 } 11, 20 -- Subject: Re: [Microscopy] viaWWW: Gelatin as the } embedding media } 11, 20 -- Message-ID: } {444F400E.3576.266CF5-at-gerd.leitinger.meduni-graz.at} } 11, 20 -- Priority: normal } 11, 20 -- In-reply-to: } {200604260129.k3Q1TCJ6016929-at-ns.microscopy.com} } 11, 20 -- X-mailer: Pegasus Mail for Windows (4.31, DE v4.31 R1) } 11, 20 -- Content-type: text/plain; charset=ISO-8859-1 } 11, 20 -- Content-description: Mail message body } 11, 20 -- X-OriginalArrivalTime: 26 Apr 2006 07:40:47.0035 (UTC) } FILETIME=[BB5540B0:01C66904] } 11, 20 -- Content-Transfer-Encoding: 8bit } 11, 20 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by } ns.microscopy.com id k3Q7el0C023530 } ==============================End of - } Headers============================== } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 8, 21 -- From amich-at-ufl.edu Wed Apr 26 11:07:52 2006 8, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3QG7pMo027838 8, 21 -- for {microscopy-at-microscopy.com} ; Wed, 26 Apr 2006 11:07:51 -0500 8, 21 -- Received: from osgjas04.cns.ufl.edu (osgjas04.cns.ufl.edu [128.227.74.134]) 8, 21 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k3QG7ntS1679594 8, 21 -- for {microscopy-at-microscopy.com} ; Wed, 26 Apr 2006 12:07:49 -0400 8, 21 -- Message-ID: {1153760693.31951146067669239.JavaMail.osg-at-osgjas04.cns.ufl.edu} 8, 21 -- Date: Wed, 26 Apr 2006 12:07:49 -0400 (EDT) 8, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 8, 21 -- To: microscopy-at-microscopy.com 8, 21 -- Subject: Re: [Microscopy] Re: viaWWW: Gelatin as the embedding media 8, 21 -- MIME-Version: 1.0 8, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 8, 21 -- X-Originating-IP: 70.152.44.187 [70.152.44.187] 8, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 8, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Thanks for all the replies - ebay was the number one response.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Once again you were most helpful. I will have to list your names in my aknowledgments ;-) Although NaHB4 could have worked, I chose to neutralize glutar with glycine, not only because it is available in the lab, but also because of the hazardous nature of NaHB4. Glycin just worked wonderfully.
Best regards,
Stephane without a "i"
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Yet another question: Due to their price and very long delays of delivery, we decided to make our formvar grids ourselves. But we have a problem: the film is full of holes! These are small holes about 50-100 nm in diameter, without sharp edges. I wondered if it did not come from traces of fine water droplets which remained on the glass slides.
Here is my protocol: I clean a glass slide by breathing on it and then rubbing with a dust-free towel. If I clean the slides with alcohol, I noticed that the film sticks to the slide and does not detach in water. Then I let the slide dry under the hood for 5 min. Then I plunge the slide in formvar (10 sec) and take it out. I let it dry in the hood for 5 min and cut the side with a razor blade. Then the usual plunge-it-in-water-praying-that-the-film-nicely-detaches-without-making-problem-thank-you-god. And so on...
Any suggestion?
Stephane-without-a-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
... holes are due to water in your formvar solution. already a very low water content causes these holes. I presume you dissolved your formvar in water-free chloroform? Did you dry (e.g. in a Speed-Vac) your (ethanol) wetted formvar slug before dissolving it?
peter heimann
nizets2-at-yahoo.com wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
I use much the same technique, so I would suspect moisture absorbed by the chloroform/other solvent.
If you're using an old bottle, poor quality or shared solvent it might be worth trying a fresh bottle of reasonable reagent quality and keep the lid on all the containers as much as possible. For instance I pour my formvar into a measuring cylinder in the fume hood and dip my slides in it one at a time. But in between dipping I have a small glass beaker that snugly fits over the neck of the measuring cylinder as a lid to reduce the risk of moisture getting into the formvar solution. I change the formvar solution when it starts to make films with too many holes.
I hope this helps.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: nizets2-at-yahoo.com
Stephane,
I usually clean my slides with acetone just before dipping them into the formvar solution. A spray each side with a spray bottle, wipe off (once) with a lint free tissue and make sure there is no acetone left on the slides. After taking the slide out of the solution I hold it in the vapour of the formvar solution for ~30 sec to make the film slightly thinner and more even. And I huff on each side of the slide after I have cut the film, to make it come off easier in the water.
Plus, something I realised in Australia, if it's a rainy or humid day (of which you have lots in Queensland, and no, this is not a joke), your film is much more likely to have holes in it. In fact, it was almost impossible to make good film on rainy days (despite aircon). Try a fine dry day for it (but then the flip side is, you have to battle the dust.... one can never win....)
Hope this helps,
Cornelia Muncke EM-Unit Physiological Laboratory University of Liverpool Crown Street Liverpool L69 3BX 0151 794 5461
} } Dear listers, } } Yet another question: } Due to their price and very long delays of delivery, } we decided to make our formvar grids ourselves. But we } have a problem: the film is full of holes! These are } small holes about 50-100 nm in diameter, without sharp } edges. I wondered if it did not come from traces of } fine water droplets which remained on the glass } slides. } } Here is my protocol: } I clean a glass slide by breathing on it and then } rubbing with a dust-free towel. If I clean the slides } with alcohol, I noticed that the film sticks to the } slide and does not detach in water. } Then I let the slide dry under the hood for 5 min. } Then I plunge the slide in formvar (10 sec) and take } it out. I let it dry in the hood for 5 min and cut the } side with a razor blade. Then the usual } plunge-it-in-water-praying-that-the-film-nicely-detaches-without- } making-problem-thank-you-god. } And so on... } } Any suggestion? } } Stephane-without-a-i }
==============================Original Headers============================== 7, 28 -- From c.muncke-at-liverpool.ac.uk Thu Apr 27 05:39:00 2006 7, 28 -- Received: from mx2.liv.ac.uk (mx2.liv.ac.uk [138.253.100.180]) 7, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3RAd0LD017986 7, 28 -- for {microscopy-at-microscopy.com} ; Thu, 27 Apr 2006 05:39:00 -0500 7, 28 -- Received: from mailhuba.liv.ac.uk ([138.253.100.36]) 7, 28 -- by mx2.liv.ac.uk with esmtp (Exim 4.54) 7, 28 -- id 1FZ3tj-0008OB-EN 7, 28 -- for microscopy-at-microscopy.com; Thu, 27 Apr 2006 11:38:59 +0100 7, 28 -- Received: from localhost ([127.0.0.1] helo=mailhuba.liv.ac.uk) 7, 28 -- by mailhuba.liv.ac.uk with esmtp (Exim 4.54) 7, 28 -- id 1FZ3tj-0005Vx-DK 7, 28 -- for microscopy-at-microscopy.com; Thu, 27 Apr 2006 11:38:59 +0100 7, 28 -- Received: from pc028200.med.liv.ac.uk ([138.253.28.200]) 7, 28 -- by mailhuba.liv.ac.uk with esmtpsa (TLSv1:RC4-SHA:128) 7, 28 -- (Exim 4.54) 7, 28 -- id 1FZ3tj-0005Vi-Cd 7, 28 -- for microscopy-at-microscopy.com; Thu, 27 Apr 2006 11:38:59 +0100 7, 28 -- Mime-Version: 1.0 (Apple Message framework v749.3) 7, 28 -- In-Reply-To: {200604270955.k3R9tS4f029758-at-ns.microscopy.com} 7, 28 -- References: {200604270955.k3R9tS4f029758-at-ns.microscopy.com} 7, 28 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 28 -- Message-Id: {1CBF07B0-5893-4094-97D1-4FFD59CE2B10-at-liverpool.ac.uk} 7, 28 -- Content-Transfer-Encoding: 7bit 7, 28 -- From: Cornelia Muncke {c.muncke-at-liverpool.ac.uk} 7, 28 -- Subject: Re: [Microscopy] holes in the formvar film 7, 28 -- Date: Thu, 27 Apr 2006 11:35:45 +0100 7, 28 -- To: microscopy-at-microscopy.com 7, 28 -- X-Mailer: Apple Mail (2.749.3) ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jaysmith8000-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, April 26, 2006 at 16:08:31 ---------------------------------------------------------------------------
Email: jaysmith8000-at-yahoo.com Name: Jay Smith
Organization: N/A
Education: Undergraduate College
Location: Los Angeles CA
Question: I'm having trouble with a homework problem. I can't find the appropriate equations needed to relate the lens used in the laser tweezer apparatus. My question is below. Any help would be greatly appreciated. Thanks!
I want to use a 5 mW He-Ne laser to trap a polystyrene bead of 8 microns in diameter (refractive index = 1.58). The bead has a density of 1050 kg/m^3. The beam source is assumed to be Gaussian, with a waist of 2 mm. You can design any lens you want, but their diameter is restricted to be not larger than 12.7mm.
Can I trap the bead? What type of lens would I need?
This Question was submitted to Ask-A-Microscopist by (scott.streiker-at-udri.udayton.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, April 26, 2006 at 19:14:36 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both scott.streiker-at-udri.udayton.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: scott.streiker-at-udri.udayton.edu Name: Scott Streiker
Organization: University of Dayton research Institute
Education: Graduate College
Location: Dayton, OH USA
Title: TEM - Imaging Atomic Lattice of Carbon
Question: Please advise protocol for imaging atomic lattice of carbon. I am trying to determine the lattice structure of carbon nano tubes (single and multiple walled). The instrument in use this study is Hitachi HRTEM Mopdel S-7600 at Accel Voltage up to 120 kV and direct magnification up to x600K.
We use formvar dissolved in ethylene dichloride (dichloroethane). If we make it ourselves than I always dry the formvar powder in an oven before using it. Often, because I am lazy, I will buy 1% formvar solution from the EM supply companies. Either works fine. We have no problems with holes but we are in a humidity controlled lab and we do keep the formvar covered as much as possible, more to prevent evaporation of the dichloroethane than concerns about water. If you are not humidity controlled than you really should wait for dry days to make films. They keep for months and years if kept dry in a desiccator.
We wash the slides by spraying with some distilled water and the gently wiping with lens paper. Let air dry for a minute or two and put into the formvar solution. We also use film casters (bottom flask container and thistle tube...formvar is pumped using a hand bulb up into the thistle tube and held there while the slide is inserted. After a few minutes the formvar is drained out and the slide is left dry in the vapor before removing. ) The film caster gives a consistently even film with large areas of the thickness desired. We regulate thickness by timing submersion in formvar and then drying time in the thistle tube. Note that ideal formvar concentration may be different if you are using the dip method rather than the film caster.
If you plan to make films regularly than I strongly recommend investing in the film caster. It's a one-time purchase and most of our users make their own films. It is convenient and much less expensive than purchasing coated grids.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
} From: {nizets2-at-yahoo.com} } Reply-To: {nizets2-at-yahoo.com} } Date: Thu, 27 Apr 2006 04:53:31 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] holes in the formvar film } } } } } ---------------------------------------------------------------------------- } 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, } } Yet another question: } Due to their price and very long delays of delivery, } we decided to make our formvar grids ourselves. But we } have a problem: the film is full of holes! These are } small holes about 50-100 nm in diameter, without sharp } edges. I wondered if it did not come from traces of } fine water droplets which remained on the glass } slides. } } Here is my protocol: } I clean a glass slide by breathing on it and then } rubbing with a dust-free towel. If I clean the slides } with alcohol, I noticed that the film sticks to the } slide and does not detach in water. } Then I let the slide dry under the hood for 5 min. } Then I plunge the slide in formvar (10 sec) and take } it out. I let it dry in the hood for 5 min and cut the } side with a razor blade. Then the usual } plunge-it-in-water-praying-that-the-film-nicely-detaches-without-making-proble } m-thank-you-god. } And so on... } } Any suggestion? } } Stephane-without-a-i } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Thu Apr 27 04:51:34 2006 } 6, 18 -- Received: from web37405.mail.mud.yahoo.com } (web37405.mail.mud.yahoo.com [209.191.87.58]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k3R9pXb6020384 } 6, 18 -- for {microscopy-at-microscopy.com} ; Thu, 27 Apr 2006 04:51:33 -0500 } 6, 18 -- Received: (qmail 1741 invoked by uid 60001); 27 Apr 2006 09:51:33 } -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-T } ransfer-Encoding; } 6, 18 -- } b=CRQz2UQbFJYYxx3xL9O/rdLuTwLYPcSY9mBW3aAbdvjBOeCrw89hflDr5PhJQKxQyVcvu2d1jqRP } Ur+4euioqL6RjlLW53DPiiCrmkzqH9KEt3Bca/WrX+fiqykrtO6WMHb7YX+Njf7ps8EJ3DvfzqPwc7 } bTwqS2RnaVi+xMZ+Y= ; } 6, 18 -- Message-ID: {20060427095133.1739.qmail-at-web37405.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via } HTTP; Thu, 27 Apr 2006 02:51:33 PDT } 6, 18 -- Date: Thu, 27 Apr 2006 02:51:33 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: holes in the formvar film } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- MIME-Version: 1.0 } 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 6, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 22 -- From dsherman-at-purdue.edu Thu Apr 27 09:51:06 2006 9, 22 -- Received: from exchange.purdue.edu (1061exfe04.adpc.purdue.edu [128.210.63.227]) 9, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3REp61G019469 9, 22 -- for {microscopy-at-microscopy.com} ; Thu, 27 Apr 2006 09:51:06 -0500 9, 22 -- Received: from EXCH02.purdue.lcl ([128.210.63.235]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 9, 22 -- Thu, 27 Apr 2006 10:51:06 -0400 9, 22 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 9, 22 -- Thu, 27 Apr 2006 14:51:04 +0000 9, 22 -- User-Agent: Microsoft-Entourage/11.2.3.060209 9, 22 -- Date: Thu, 27 Apr 2006 10:51:04 -0400 9, 22 -- Subject: Re: [Microscopy] holes in the formvar film 9, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 9, 22 -- To: {nizets2-at-yahoo.com} , "message to: MSA list" {microscopy-at-microscopy.com} 9, 22 -- Message-ID: {C0765298.F5E9%dsherman-at-purdue.edu} 9, 22 -- Thread-Topic: [Microscopy] holes in the formvar film 9, 22 -- Thread-Index: AcZqCgHbQFXdeNX9EdqsawARJN08Mg== 9, 22 -- In-Reply-To: {200604270953.k3R9rVJd024493-at-ns.microscopy.com} 9, 22 -- Mime-version: 1.0 9, 22 -- Content-type: text/plain; 9, 22 -- charset="US-ASCII" 9, 22 -- Content-transfer-encoding: 7bit 9, 22 -- X-OriginalArrivalTime: 27 Apr 2006 14:51:06.0142 (UTC) FILETIME=[032213E0:01C66A0A] ==============================End of - Headers==============================
Well there seems to be a general agreement that it is probably water contamination. This is comforting if I know the cause of the problem, but I still have to find a solution to it because:
- I bought the formvar solution ready to use in dichloroethane - I use it under the hood (of course) and close it between the slide (I prepare 2 slides at a time ;-)) - we are working with air conditioning, at controlled 60% of humidity. - The weather was very nice these last days: very sunny and 24°C.
Thank you for your comments, I will find a way!
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
One possibility that hasn't been mentioned is that if the formvar solution is colder than ambient, then when you pull the slide out of the solution some water vapor will condense on it. This is especially true on humid days, etc. Incidentally, chilling the formvar solution prior to breathing on the newly-cast films will ensure a large number of holes...
Andy Bowling
==============================Original Headers============================== 2, 19 -- From abowling-at-mail.utexas.edu Thu Apr 27 11:23:06 2006 2, 19 -- Received: from ironwood.mail.utexas.edu (ironwood.mail.utexas.edu [128.83.32.56]) 2, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3RGN5Ys007866 2, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 27 Apr 2006 11:23:06 -0500 2, 19 -- Received: from wb8-a.mail.utexas.edu ([128.83.126.148]) 2, 19 -- by ironwood.mail.utexas.edu with ESMTP; 27 Apr 2006 11:23:05 -0500 2, 19 -- Received: (qmail 11666 invoked from network); 27 Apr 2006 16:23:05 -0000 2, 19 -- Received: from dhcp-146-6-151-204.biosci.utexas.edu (HELO ?146.6.151.204?) (abowling-at-146.6.151.204) 2, 19 -- by wb8.mail.utexas.edu with (RC4-MD5 encrypted) ESMTPSA; 27 Apr 2006 16:23:05 -0000 2, 19 -- Message-ID: {4450EF8B.8050904-at-mail.utexas.edu} 2, 19 -- Date: Thu, 27 Apr 2006 11:21:31 -0500 2, 19 -- From: Andrew Bowling {abowling-at-mail.utexas.edu} 2, 19 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 2, 19 -- X-Accept-Language: en-us, en 2, 19 -- MIME-Version: 1.0 2, 19 -- To: Microscopy-at-microscopy.com 2, 19 -- Subject: [Microscopy] Re: holes in the formvar film 2, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I had heaps of trouble with my formvar for a while. I'm in Vancouver, so rain is a fact of life here, especially in the winter. After trying everything I could think of (only doing it on dry days, fume hoods, different dipping methods and conatiners, buying new formvar solution) I finally made my own formvar from our stock powder in the lab (in dichloroethane) and have never had a problem since. Now I make the solution myself, throw it out if it gets to be 6 months old, or if it has been opened more than 5 times. Its possible that this is a tad over zealous, but it works for me.
Good luck. Robin
Robin Elizabeth Young, M.Sc. PhD Candidate Dept. of Botany, University of British Columbia 6270 University Blvd Vancouver, BC V6T 1Z4 Fax: 604-822-6089
On 27-Apr-06, at 8:57 AM, nizets2-at-yahoo.com wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Well there seems to be a general agreement that it is } probably water contamination. This is comforting if I } know the cause of the problem, but I still have to } find a solution to it because: } } - I bought the formvar solution ready to use in } dichloroethane } - I use it under the hood (of course) and close it } between the slide (I prepare 2 slides at a time ;-)) } - we are working with air conditioning, at controlled } 60% of humidity. } - The weather was very nice these last days: very } sunny and 24°C. } } Thank you for your comments, I will find a way! } } Stéphane }
==============================Original Headers============================== 10, 31 -- From youngre-at-interchange.ubc.ca Thu Apr 27 12:01:50 2006 10, 31 -- Received: from mta2.mail-relay.ubc.ca (mta2.mail-relay.ubc.ca [137.82.45.4]) 10, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3RH1nP5018751 10, 31 -- for {microscopy-at-microscopy.com} ; Thu, 27 Apr 2006 12:01:49 -0500 10, 31 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 10, 31 -- by mta2.mail-relay.ubc.ca (8.12.11/8.12.11) with ESMTP id k3RH1Ypa019993 10, 31 -- for {microscopy-at-microscopy.com} ; Thu, 27 Apr 2006 10:01:34 -0700 (PDT) 10, 31 -- (envelope-from youngre-at-interchange.ubc.ca) 10, 31 -- Received: from [137.82.136.151] (samuelsg5.botany.ubc.ca [137.82.136.151]) 10, 31 -- by smtp.interchange.ubc.ca 10, 31 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 10, 31 -- with ESMTPSA id {0IYE00EJH4ML6D-at-smtp.interchange.ubc.ca} for 10, 31 -- microscopy-at-microscopy.com; Thu, 27 Apr 2006 10:01:34 -0700 (PDT) 10, 31 -- Date: Thu, 27 Apr 2006 10:01:33 -0700 10, 31 -- From: Robin Young {youngre-at-interchange.ubc.ca} 10, 31 -- Subject: Re: holes in the formvar film 10, 31 -- In-reply-to: {200604271557.k3RFvl8x032682-at-ns.microscopy.com} 10, 31 -- To: microscopy-at-microscopy.com 10, 31 -- Cc: nizets2-at-yahoo.com 10, 31 -- Message-id: {4BDA715F-0B39-4028-92A8-BDD4BAF5F066-at-interchange.ubc.ca} 10, 31 -- MIME-version: 1.0 (Apple Message framework v749.3) 10, 31 -- X-Mailer: Apple Mail (2.749.3) 10, 31 -- Content-type: text/plain; format=flowed; delsp=yes; charset=ISO-8859-1 10, 31 -- References: {200604271557.k3RFvl8x032682-at-ns.microscopy.com} 10, 31 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.4.27.85105 10, 31 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 10, 31 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P1_5 0, __C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0, __STOCK_PHRASE_6 0 10, 31 -- X-Spam-Level: 10, 31 -- X-Spam-Flag: No 10, 31 -- Content-Transfer-Encoding: 8bit 10, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3RH1nP5018751 ==============================End of - Headers==============================
On Apr 27, 2006, at 8:56 AM, nizets2-at-yahoo.com wrote:
} If I clean the slides } with alcohol, I noticed that the film sticks to the } slide and does not detach in water. } } Well there seems to be a general agreement that it is } probably water contamination. This is comforting if I } know the cause of the problem, but I still have to } find a solution to it because: } } - I bought the formvar solution ready to use in } dichloroethane } - I use it under the hood (of course) and close it } between the slide (I prepare 2 slides at a time ;-)) } - we are working with air conditioning, at controlled } 60% of humidity. } - The weather was very nice these last days: very } sunny and 24°C. } } Thank you for your comments, I will find a way! } Dear Stephane, So far no one has mentioned how to get formvar films off of alcohol-cleaned slides. Our method is to clean the slides with ethanol, then apply a thin film of oil to the slides by rubbing the sides of your nose with your fingers and transferring the oil from your skin to the slide. An alternative is to dissolve Apiezon L in petroleum ether and dip the slide in that solution. In any case, the trick is to get the slide controllably dirty. When removing the film we use warm water, and when using the Apiezon, we add 0.25 g of Alconox to 1 L of water. 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============================== 5, 23 -- From tivol-at-caltech.edu Thu Apr 27 14:10:55 2006 5, 23 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k3RJAtDG030288 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Thu, 27 Apr 2006 14:10:55 -0500 5, 23 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 5, 23 -- by fire-ox-postvirus (Postfix) with ESMTP id B7D57363E4 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Thu, 27 Apr 2006 12:10:54 -0700 (PDT) 5, 23 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 5, 23 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id 62DD834BE6 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Thu, 27 Apr 2006 12:07:04 -0700 (PDT) 5, 23 -- Mime-Version: 1.0 (Apple Message framework v623) 5, 23 -- In-Reply-To: {200604271556.k3RFuRH5030392-at-ns.microscopy.com} 5, 23 -- References: {200604271556.k3RFuRH5030392-at-ns.microscopy.com} 5, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 23 -- Message-Id: {e4246a85769dab4b06ecf9d68e844a34-at-caltech.edu} 5, 23 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 23 -- Subject: Re: [Microscopy] Re: holes in the formvar film 5, 23 -- Date: Thu, 27 Apr 2006 12:17:10 -0700 5, 23 -- To: microscopy-at-msa.microscopy.com 5, 23 -- X-Mailer: Apple Mail (2.623) 5, 23 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k3RJAtDG030288 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both fishon-at-umich.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: fishon-at-umich.edu Name: Chris A. Edwards
Organization: University of Michigan, Microscopy and Image-analysis Lab
Title-Subject: [Filtered] Releasing Formvar Films
Question: Hi Listers, For those of you having problems with formvar film sticking to glass slides, may I refer you to the following article: "A Technique for Achieving Consistent Release of Formvar Films from Clean Glass Slides"; J.Electron Microscopy Technique, 1:203-204, 1984.