Over the Christmas break we routinely shut our microscopes down to run only on ion pumps as our cooling water can be a little "temperamental".
This year we had a power glitch which turned the microscopes completely off. To top it off our cooling water is out of action for maintenance until next week!! I was hoping to get all the ion pumps running on the microscopes to keep the vacuums at leat a bit clean and I have managed to get this done on all but our JEOL 2010. Does anyone know if it is possible to get just the ion pumps running without needing cooling water on a 2010?
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==============================Original Headers============================== 17, 28 -- From Colin.Veitch-at-csiro.au Mon Jan 2 18:38:05 2006 17, 28 -- Received: from vic-ironport-ext-out1.csiro.au (vic-ironport-ext-out1.csiro.au [150.229.64.37]) 17, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k030c3V4024434 17, 28 -- for {microscopy-at-msa.microscopy.com} ; Mon, 2 Jan 2006 18:38:04 -0600 17, 28 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=xU3RCN8AXAmWEvu85jV1wrWTuUHnFiSlBXAaGGnTue1S/WJESDcqcwHjBKBfIWsOg4hoNydsUlK2UaXg0lctHoNuIRakZsSu0pTgT/UHQM1uofzGf4egwYNOnYE3LgVO; 17, 28 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 17, 28 -- by vic-ironport-ext-out1.csiro.au with ESMTP; 03 Jan 2006 11:38:02 +1100 17, 28 -- X-IronPort-AV: i="3.99,323,1131282000"; 17, 28 -- d="scan'208"; a="63973513:sNHT23878160" 17, 28 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 17, 28 -- Tue, 3 Jan 2006 11:38:02 +1100 17, 28 -- content-class: urn:content-classes:message 17, 28 -- MIME-Version: 1.0 17, 28 -- Content-Type: text/plain; 17, 28 -- charset="us-ascii" 17, 28 -- Subject: Restarting a JEOL 2010 on ion pumps 17, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 17, 28 -- Date: Tue, 3 Jan 2006 11:38:02 +1100 17, 28 -- Message-ID: {32CDDDAA7161394599F0025494915749051DD3-at-exvic5-gex.nexus.csiro.au} 17, 28 -- X-MS-Has-Attach: 17, 28 -- X-MS-TNEF-Correlator: 17, 28 -- Thread-Topic: Restarting a JEOL 2010 on ion pumps 17, 28 -- Thread-Index: AcYP/fQ3/LdcT+y2Rj221MWzoKvg+g== 17, 28 -- From: {Colin.Veitch-at-csiro.au} 17, 28 -- To: {microscopy-at-msa.microscopy.com} 17, 28 -- X-OriginalArrivalTime: 03 Jan 2006 00:38:02.0959 (UTC) FILETIME=[F47CA5F0:01C60FFD] 17, 28 -- Content-Transfer-Encoding: 8bit 17, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k030c3V4024434 ==============================End of - Headers==============================
FOCUS ON MICROSCOPY 2006, Perth, Australia, April 9-12, 2006 19th International Conference on 3D Image Processing in Microscopy 18th International Conference on Confocal Microscopy
Dear Colleagues
January 9, 2006, the deadline for abstract submission for the Perth conference is nearing.
Please submit your abstract by that date.
The program for the conference will be finalized and available on our website on Jan 23, 2006. Authors will also be informed individually by E-mail on the placement of their contribution in the program. Abstracts for oral and poster presentations are sollicited. Submission preferably through the conference website: http://focusonmicroscopy.org/ where also the conference registration is open and hotel booking information is available.
The earlier than usual deadline of 9 Jan. was chosen to give delegates sufficient time -after the program has been announced- to arrange their travel and acccommodation before the conference starting date of Sunday 9 April, 2006.
Please note that the conference will take place in the Esplanade Hotel in Fremantle, the historic waterfront suburb of Perth, See further our website.
The program will start on Sunday April 9, around 18 hours with an opening symposium in the Esplanade hotel followed by a welcome reception.
The conference Focuson Microscopy 2006 will take place as the next in a series of unique interdisciplinary meetings on advanced multidimensional light microscopy and image processing. The conference will be hosted by the University of Western Australia in Perth.
Focus on Microscopy 2006 is the continuation of a conference series presenting the latest innovations in optical microscopy and its applications in biology, medicine, material science, and information storage. 3D optical imaging and related theory are traditional subjects for the conference. The conference series is also known for covering the rapid development of advanced fluorescence labeling techniques for the confocal and multi-photon 3D imaging of -live- biological specimens. This year, in addition, special attention will be given to imaging in thick tissues.
Abstracts for contributions are invited and can now be submitted through the website:
www.FocusOnMicroscopy.org
where further information on the present and previous FOM conferences can be found.
Important dates:
Deadline for the submission of abstracts: January 9, 2006 Draft program available on the web: January 23, 2006 at website www.FocusOnMicroscopy.org Deadline for early registration: February 20, 2006
Welcoming you to beautiful Perth for the FOM2006 conference and exhibition.
With the best wishes for the New Year and on behalf of the organizing committee,
David Sampson, University of Western Australia, Perth, Australia
Fred Brakenhoff Swammerdam Institute for Life Sciences, University of Amsterdam, The Netherlands
E-mail: info2006-at-FocusOnMicroscopy.org
Web: www.FocusOnMicroscopy.org
==============================Original Headers============================== 39, 30 -- From brakenho-at-science.uva.nl Tue Jan 3 02:18:50 2006 39, 30 -- Received: from imap.science.uva.nl (imap.science.uva.nl [146.50.4.51]) 39, 30 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k038IluR006028 39, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 3 Jan 2006 02:18:48 -0600 39, 30 -- Received: from webmail.science.uva.nl [146.50.4.91] 39, 30 -- by imap.science.uva.nl with ESMTP (sendmail 8.11.6p2/config 11.36). 39, 30 -- id k038Ij231650; Tue, 3 Jan 2006 09:18:46 +0100 39, 30 -- Received: from localhost 39, 30 -- by webmail.science.uva.nl (sendmail 8.11.6/config 11.35). 39, 30 -- id k038Ij232296; Tue, 3 Jan 2006 09:18:45 +0100 39, 30 -- X-Organisation: Faculty of Science, University of Amsterdam, The Netherlands 39, 30 -- X-URL: http://www.science.uva.nl/ 39, 30 -- Received: from localhost ([127.0.0.1]) 39, 30 -- (SquirrelMail authenticated user brakenho); 39, 30 -- by webmail.science.uva.nl with HTTP; 39, 30 -- Tue, 3 Jan 2006 09:18:45 +0100 (CET) 39, 30 -- Message-ID: {51496.84.254.54.91.1136276325.squirrel-at-84.254.54.91} 39, 30 -- Date: Tue, 3 Jan 2006 09:18:45 +0100 (CET) 39, 30 -- Subject: FOM2006 abstract deadline 9 Jan. near! FocusOnMicroscopy, Perth, 39, 30 -- April 9-14 2006 39, 30 -- From: "Fred Brakenhoff" {brakenho-at-science.uva.nl} 39, 30 -- To: microscopy-at-msa.microscopy.com 39, 30 -- User-Agent: SquirrelMail/1.4.3a 39, 30 -- X-Mailer: SquirrelMail/1.4.3a 39, 30 -- MIME-Version: 1.0 39, 30 -- Content-Type: text/plain;charset=iso-8859-1 39, 30 -- Content-Transfer-Encoding: 8bit 39, 30 -- X-Priority: 3 (Normal) 39, 30 -- Importance: Normal 39, 30 -- X-Virus-Scanned: by amavisd-new ==============================End of - Headers==============================
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Email: fisiltd-at-gmail.com Name: Howard Freeman
Organization: FISI Ltd
Title-Subject: [Filtered] O'Brien & McCully, Study of Plant Structure
Question: I have a copy of The study of Plant Structure, O'Brien & McCully, first printing, 1981 in very good condition. I used this for my teaching but have moved away from histology. If anyone is interested please send email to fisiltd-at-gmail.com
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Organization: pos-doc at INSA - Institut National des Sciences AppliquČes de Lyon
Title-Subject: [Filtered] LM on 304L grey superficial layer and black lines
Question: I have shot peening 304L steel specimens, thickness about 5mm. Objective to visualise martensitic phase. I used tradional mechanical polishing, till 1200 grade, different etching based on water, HCl and potassium dissulfite (Berahas's tint). I did not clearly observe a martensitic layer but some insland, there is a dark grey to black layer on treated surface of about 10 microns, in case of highest resolution 200x I see it bright grey. What you believe it to be? I observe black flowing lines, variable lengths from 5 to 200 microns , paralel to surface through all specimen thickness and all its length. What it cab be? Some mention they are mechanical polishing effect others ferrite phase.
Thanks for whom took time reading or answering! Best new year wishes! Soraia Pirfo
Hi Sue, You give me an occasion to offer via the listserver some lore on MnO4 staining and epoxy resin adaptations to MnO4 staining that I provided off-listserver earlier this year in 2 other messages. I'll offer my message in three lengths.
SHORT VERSION is: Using epoxy embedding that excludes MNA (MNA?), MnO4 section staining followed by a lead stain (Sato's) can give fantastic contrast to everything --- not just membranes --- including any dirt/oil collected from the trough when picking up the sections, or rinsed off the pipette tip when dispensing drops of stain. The contrast is even greater if tannic acid has been used in the fix. Section staining of Epon-DDSA or Spurr embeddings is OK, but Araldite gives distinctly finer-grain higher-resolution staining.
MEDIUM VERSION: NMA renders use of KMnO4 as a section stain impossible; MnO4 reacts with the cured resin. Reedy, J Cell Biol 1965 26:309-11
We prefer the Mn-Pb sequence for higher contrast generally, and especially in the thinnest sections - 15-30 nm. So we gave up NMA about 30 years ago, and just heretically fiddled the resin-DDSA proportions until we got a hardness we liked; it worked just fine. With Epon that was that. We prefer Araldite because it is less even reactive than Epon or Spurr with MnO4. With Araldite-DDSA, we use Araldite 506. We found the best mixture still a bit too brittle, so added a bit of DER 736 as a "plasticizer", have used that for over 30 years. Araldite 506 50g 10g DDSA 75g 15g DER 736 10g 2g 1.8% v/v DMP-30 (range 1.5% to 2.0%). Reference Reedy et al (1983) J. Muscle Res. Cell Motil. 4:25-53.
LONG LONG VERSION (data dump): (This comes mostly from my off-listserver reply to some contributors to NetNotes in the May or June 2005 Microscopy today, regarding a query by Ursula J Potter about MnO4 use as a section stain.)
I have used 1-2% MnO4 section staining since 1964, almost exclusively since 1970, always followed by Pal's bleach (see below) to eliminate surface "pepper" of MnO2 ppt, followed then by a lead stain, of which Sato's appeals to us as the best and most storage-stable (Hanaichi et al, 1986, Proc 11th Internat'l EM Congress (Kyoto), p 2181). After publishing my brief note on the MNA problem (J Cell Biol, 1965, 26:309), I soon switched completely to Araldite-DDSA embedding, because it turned out that cured Araldite was much less reactive with MnO4 than any other epoxy resin mix I ever tested, including Spurr's when it later appeared. I used an extreme TEST, exposing cured blocks of various embedding resin mixtures to HOT KMnO4 solution (test tube immersed in boiling water) for 1 hr. In that test, the Epon-DDSA resin "preferred" in my Brief Note developed a blackened micro-fissured tree-bark surface (like that of MNA-Epon in unheated KMnO4, while the Araldite-DDSA resin was scarcely discolored at all. The Epon-DDSA sections typically presented a more nano-granular staining than did Araldite sections.
I think I gave up using barium MnO4 because it seemed to give blotchy staining more often than KMnO4. I also guessed that barium ion might capture carbonate as Pb did; Pb carbonate was the ubiquitous contaminant from hell with all the early lead stains that preceded lead citrate.
We wanted the maximum staining contrast that is provided by the Mn-Pb sequence because we mainly use 15-30 nm sections. Adding in UrAc to the sequence helps not at all. We do use UrAc as a block-stain, or as a secondary post-fixative (preferred to OsO4 in my lab) when our primary fixative is tannic acid with or (usually) without glutaraldehyde (TA-URAC works beautifully on myofilaments and on residual membranes in our permeabilized demembranated insect muscle, both in aqueous buffers at room temperature and as freeze-subst'n chemistry in acetone at -80 to -90°C). Recently, we have even found that the actin monomer substructure of thin myofilaments can be seen-- it appears to be relatively negatively stained, coated with metal derived from the TAURAC and Mn-Pb sequences (Fig 3, blue outlined: Fig 5 B; Liu et al, J Struct Biol, 2004, 147:268-282).
We had to experiment to find the optimum staining times for KMnO4, Pal's bleach washing, and lead stain needed to saturate contrast without "digesting" or extracting structures out of the sections. We learned by deliberately over-treating sections with each step. 60-90 min will remove Z-bands and (maybe) myofilaments from 100-150 nm Araldite sections, leaving a negative image, actually a cavity in the resin. Staining saturatees slower in Araldite than in Epon sections. 100 nm sections become stain-saturated slower than 25 nm sections. Cross-sections of muscle saturate much faster than longitudinal sections. 2-10 min in MnO4, 10-30s washing in Pal's bleach 1:100, and 1-5 min on Sato's Pb stain works for 25 nm longitudinal sections of muscle.
Stain apparently penetrates the section only where the pure embedding medium is modified by the presence of stained structures that intercept the section surface. The stains percolate only through the embedded structures, NOT through blank empty resin regions, as we found when sections had piled up or folded two-deep on one another; the covered section (sandwiched between carbon support film and an overlying surface section) would appear stained only within 0.1-0.2 um laterally from wherever the overlying section contained embedded structures such as filaments or membranes to conduct stain from the solution to the underlying structures in a covered section.
PAL's BLEACH, from the J. D. Robertson 1963 paper cited in my 1965 brief note: Distilled water 100 ml Potassium sulphite 0.5 g Oxalic acid 0.5 g dilute 1:100 for use (3-4 drops of above stock in 10 ml water), and run 10-20 drops over the grid after MnO4 staining. Water wash immediately after MnO4 and after Pal's.
All our staining is done by floating grids on small spherical drops of stain on fresh-washed Parafilm. (I'm not familiar with the vertical-insertion immersion staining of grids mentioned by Dr W. Muss in his helpful reply to you Dec 30.) Keys to clean staining: draw up the stains from under the surface in the stock bottles, discard the first 3-6 drops before putting the drop to be used down on a freshly washed Parafilm surface. Mistrust paranoically all unwashed surfaces on forceps and parafilm and pipette barrels as likely to have finger grease that will get on your sections or support films and become intensely stained contaminants. Wash everything you mistrust that will contact sections and grids with lavish flows and jets of deionized or distilled water. Stock lead stain needs replacing when it starts to "fails" in producing contrast, as often as every 4 weeks. Stock 2% KMnO4 lasts "forever"; just remember to respect and go below its MnO2-dirty surface when filling your pipette.
Around 1970 I switched to a lower-viscosity Araldite 506 mix (with DDSA, DER 732, DMP-30: see MEDIUM VERSION above for recipe and reference) that we have used ever since. It becomes as water-thin in viscosity as Spurr's when heated to 60°C, so we rotate vials on a small tilted platform in the oven for 20' x 3 changes to optimize infiltration, bypassing the 50:50 (etc) intermediate solvent-resin mixtures to go directly from solvent into 100% accelerated resin mix. We soon abandoned some of the other old-time religion as well, giving up propylene oxide completely to go directly from acetone or ethanol, after we tried deliberately curing 10-15 ml vials of accelerated resin with 10-20% solvent thoroughly blended in; the PO additive produced a much "cheesier" final cured resin than equivalent percentage of acetone or EtOH. (However, see my Dec 6 2005 on-listserver message RE: LR White contrast, calling attention to SW Brorson's use of propylene oxide and/or high [accelerator] to improve exposure of antigens in epoxy sections to immunolabeling). We also explored the epoxy-anhydride ratio over a wide and quite heretical range (30-70% v/v anhydride) to finally settle on a "nicer" final cured resin (less brittle during block-trimming than the gospel ratio) because we found acceptable cutting, staining, and structural preservation over the whole range we tried.
Some issues I consider still unfinished: 1) Can shorter or colder (etc) staining of Epon(replacement)-DDSA or Spurr sections eliminate the nano-granularity to approach the finer-grained stain we routinely get in Araldite? 2) Does our Mn-Pb section stain perhaps "clog" or "fill in" some fine structures in the 5 nm and smaller range that are more delicately and clearly brought out by the Ur-Pb staining sequence? I've yet to re-check my impression that the 4.8 nm axial repeat on thick filaments routinely showed up 30 years ago on optical diffraction of insect muscle section EMs, before we started using the MnPb exclusively; and that it never shows up now that we no longer use the weaker contrasting UrPb sequence.
-mike reedy-
At 9:28 PM -0600 12/29/05, susan.vanhorn-at-stonybrook.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
The easiest way to get a transmitted light image on the 510 is to just click on ChD, which is a transmitted light detector. This will collect all the light that passes through your sample AT THE SAME TIME you are collecting confocal fluorescent images. So no need to double-expose your sample. You will still need to set the condenser and neutral density filters, etc., for Koehler.
I am not sure if it is a bug that the HAL light goes off when scanning - I think it is probably a protection (safety) so the PMTs do not get blasted.
-Holly __________________ Holly L. Aaron CRL Molecular Imaging Center http://imaging.berkeley.edu
-----Original Message----- X-from: lubo-at-berkeley.edu [mailto:lubo-at-berkeley.edu] Sent: Wednesday, December 28, 2005 6:46 AM To: hollya-at-berkeley.edu
Thanks guys, I really apreciate it. I will try some of the mentioned methods and I will update you with the results and I will tell you which method(s) worked with me.
Best Regards
Hany
On 12/30/05, ramadanhany-at-gmail.com {ramadanhany-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 } ---------------------------------------------------------------------------- } } Good morning, } } I grow tantalum oxide electrochemically on tantalum substrates, the } issue is that using SEM I can say that the surface of tantalum oxide } is full of defects "pin holes". I just use mechanical polishing of } tantalum before growing oxide, so I think that these defects result } from impurities of polishing sand papers. Is there any other way I can } polish tantalum to the finest grade avoiding these impurities? } } I appreciate your responses. } } Thanks } -- } ********************************************************** } Hany Ramadan } Graduate student } Chemistry department } McMaster university, Hamilton, Ontario, Canada } 905-525-9140 x: 26322 } elsayeh-at-mcmaster.ca } ********************************************************** } } } ==============================Original Headers============================== } 5, 23 -- From ramadanhany-at-gmail.com Fri Dec 30 10:10:44 2005 } 5, 23 -- Received: from zproxy.gmail.com (zproxy.gmail.com [64.233.162.201]) } 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id jBUGAiMs015284 } 5, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 30 Dec 2005 10:10:44 -0600 } 5, 23 -- Received: by zproxy.gmail.com with SMTP id o37so2448119nzf } 5, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 30 Dec 2005 08:10:44 -0800 (PST) } 5, 23 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 5, 23 -- s=beta; d=gmail.com; } 5, 23 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; } 5, 23 -- b=PYN5psgGfgI0WLNifAGGkP00f9GUSHXDkWrIi+bDVwtHrgVUiPHtTSNzH8/qCj4iWo1PnzfQMBQZCsCEzGDhrrwV0seSVvIrg57GrNfpfp4NrW/hMtWkQ+jXcNmfpDmh6jp5Cvjb4kwfB1DUnbR47y1HRX3kvEdsyqmEtn0yvqE= } 5, 23 -- Received: by 10.65.231.8 with SMTP id i8mr4207763qbr; } 5, 23 -- Fri, 30 Dec 2005 08:10:44 -0800 (PST) } 5, 23 -- Received: by 10.65.156.18 with HTTP; Fri, 30 Dec 2005 08:10:44 -0800 (PST) } 5, 23 -- Message-ID: {8d8ce5a30512300810o3ce7e534tc4e1ac19820b4cd4-at-mail.gmail.com} } 5, 23 -- Date: Fri, 30 Dec 2005 11:10:44 -0500 } 5, 23 -- From: Hany Ramadan {ramadanhany-at-gmail.com} } 5, 23 -- To: Microscopy-at-microscopy.com } 5, 23 -- Subject: Tantalum polishing } 5, 23 -- MIME-Version: 1.0 } 5, 23 -- Content-Type: text/plain; charset=ISO-8859-1 } 5, 23 -- Content-Disposition: inline } 5, 23 -- Content-Transfer-Encoding: 8bit } 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id jBUGAiMs015284 } ==============================End of - Headers============================== }
-- ********************************************************** Hany Ramadan Graduate student Chemistry department McMaster university, Hamilton, Ontario, Canada 905-525-9140 x: 26322 elsayeh-at-mcmaster.ca **********************************************************
==============================Original Headers============================== 7, 25 -- From ramadanhany-at-gmail.com Tue Jan 3 18:28:12 2006 7, 25 -- Received: from zproxy.gmail.com (zproxy.gmail.com [64.233.162.193]) 7, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k040SCVw032542 7, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 3 Jan 2006 18:28:12 -0600 7, 25 -- Received: by zproxy.gmail.com with SMTP id n29so4190029nzf 7, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 03 Jan 2006 16:28:12 -0800 (PST) 7, 25 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 25 -- s=beta; d=gmail.com; 7, 25 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 7, 25 -- b=jqvEITCtrHYGz22VW5v2TzDwTUPG6E7u8wxIz49Ct4T5ydsjuRU2DFVPLQemUrhx6KfdA71OlVgOpsJXvgrRMzHZ4Kd65rnXBquoj10M2tR4eawDB8YJdyC2xyIerEuZ1tzCcVsmxF6E6l/77PdQGmNx/sh55aJ32NRuNqyfplM= 7, 25 -- Received: by 10.65.177.14 with SMTP id e14mr4536714qbp; 7, 25 -- Tue, 03 Jan 2006 16:28:11 -0800 (PST) 7, 25 -- Received: by 10.65.156.18 with HTTP; Tue, 3 Jan 2006 16:28:11 -0800 (PST) 7, 25 -- Message-ID: {8d8ce5a30601031628o1cb70a8bre850d04cfd3b18d0-at-mail.gmail.com} 7, 25 -- Date: Tue, 3 Jan 2006 19:28:11 -0500 7, 25 -- From: Hany Ramadan {ramadanhany-at-gmail.com} 7, 25 -- To: Microscopy-at-microscopy.com 7, 25 -- Subject: Re: [Microscopy] Tantalum polishing 7, 25 -- In-Reply-To: {200512301648.jBUGmvLq022871-at-ns.microscopy.com} 7, 25 -- MIME-Version: 1.0 7, 25 -- Content-Type: text/plain; charset=ISO-8859-1 7, 25 -- Content-Disposition: inline 7, 25 -- References: {200512301648.jBUGmvLq022871-at-ns.microscopy.com} 7, 25 -- Content-Transfer-Encoding: 8bit 7, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k040SCVw032542 ==============================End of - Headers==============================
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Title-Subject: [Filtered] SEMS annual meeting for 2006
Question: This year the annual meeting for the Southeastern Microscopy Society (SEMS) will be held jointly with the Association of Southeastern Biologists (ASB). The meeting will be March 29-April 1 in Gatlinburg, Tennessee at the Gatlinburg Convention Center. This event brings together approximately 800 biologists from across the southeastern United States. Interests are diverse, but range from genetics and molecular biology, to physiology and population biology, to community and ecosystem ecology.
Registration deadline is January 17, 2006 and information is available at the SEMS website:
http://www.semicroscopy.org/index.html or http://www.semicroscopy.org/meetings/call-for-papers.html
Cynthia S. Goldsmith Secretary, Southeastern Microscopy Society (SEMS) Mailstop G32 Centers for Disease Control and Prevention (CDC) Atlanta, GA 30333
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Title-Subject: [Filtered] Comparing cryo- and cryo-immuno diamond knives
Question: I would appreciate an advice from people who use cryo-immuno diamond knife from Diatome for the Tokuyasu technique. It is a newer model than their cryo diamond knife. Both can be seen at the following web site. http://www.emsdiasum.com/diatome/knife/immunocytochemistry.htm Is the newer model indeed better than the older one? Is there somebody in San Diego area who uses the cryo immuno knife? Thank you very much, Halina
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Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers Institute
Title-Subject: [Filtered] glue to make ribbon's stick together
Question: Hello, I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions. Thanks in adance. Rhonda Allen rra-at-stowers-institute.org
I seem to recall that diluted rubber cement can be used
rra-at-stowers-institute.org wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } 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 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] glue to make ribbon's stick together } } Question: Hello, } I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions. } Thanks in adance. } Rhonda Allen } rra-at-stowers-institute.org } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Tue Jan 3 20:11:11 2006 } 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 6, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k042BADB010462 } 6, 12 -- for {microscopy-at-microscopy.com} ; Tue, 3 Jan 2006 20:11:10 -0600 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110402bfe0df2a3482-at-[206.69.208.22]} } 6, 12 -- Date: Tue, 3 Jan 2006 20:11:08 -0600 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: rra-at-stowers-institute.org (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: glue to make ribbon's stick together } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
-- Greg Erdos 5410 SE 185th Ave. Micanopy, FL 32667
==============================Original Headers============================== 3, 24 -- From gwe-at-ufl.edu Wed Jan 4 16:03:58 2006 3, 24 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 3, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k04M3v0Z007902 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 4 Jan 2006 16:03:58 -0600 3, 24 -- Received: from [192.168.1.97] (adsl-152-56-36.gnv.bellsouth.net [70.152.56.36]) 3, 24 -- (authenticated bits=0) 3, 24 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k04M3tuh1663198; 3, 24 -- Wed, 4 Jan 2006 17:03:56 -0500 3, 24 -- Message-ID: {43BC4650.1010103-at-ufl.edu} 3, 24 -- Date: Wed, 04 Jan 2006 17:04:00 -0500 3, 24 -- From: greg erdos {gwe-at-ufl.edu} 3, 24 -- Reply-To: gwe-at-ufl.edu 3, 24 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 3, 24 -- MIME-Version: 1.0 3, 24 -- To: rra-at-stowers-institute.org, microscopy-at-microscopy.com 3, 24 -- Subject: Re: [Microscopy] viaWWW: glue to make ribbon's stick together 3, 24 -- References: {200601040223.k042NquA014469-at-ns.microscopy.com} 3, 24 -- In-Reply-To: {200601040223.k042NquA014469-at-ns.microscopy.com} 3, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 24 -- Content-Transfer-Encoding: 7bit 3, 24 -- X-Spam-Status: hits=-1.524, required=5, tests=BAYES_01 3, 24 -- X-UFL-Spam-Status: hits=-1.524, required=5, tests=BAYES_01 3, 24 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 24 -- 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 --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both tsoumt-at-ilan.tpg.gov.tw as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: tsoumt-at-ilan.tpg.gov.tw Name: Jack Tsou
Organization: I-lan Hospital, Taiwan
Title-Subject: [Filtered] Has anyone successfully mounted Canon pro1 onto trinocular port of Olympus BX51?
Question: I'm quite interested in the digital camera Canon pro1 and would like to know the detail if anybody had ever successfully mounted it onto the trinocular port of BX51. Thanks in advance!
If you haven't committed to the Canon Powershot Pro1 yet, I strongly recommend you move up to a camera with a removable lens like the Canon EOS 350D or The Nikon D50 - and use the Microscope as the lens.
On 5 Jan 2006, at 19:57, tsoumt-at-ilan.tpg.gov.tw 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 } ---------------------------------------------------------------------- } ----- Remember this posting is most likely not from a Subscriber, so } when replying please copy both tsoumt-at-ilan.tpg.gov.tw as well as } the MIcroscopy Listserver } ---------------------------------------------------------------------- } ----- } } Email: tsoumt-at-ilan.tpg.gov.tw } Name: Jack Tsou } } Organization: I-lan Hospital, Taiwan } } Title-Subject: [Filtered] Has anyone successfully mounted Canon pro1 } onto trinocular port of Olympus BX51? } } Question: I'm quite interested in the digital camera Canon pro1 and } would like to know the detail if anybody had ever successfully mounted } it onto the trinocular port of BX51. Thanks in advance! } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== 6, 12 -- From } zaluzec-at-microscopy.com Thu Jan 5 17:20:05 2006 6, 12 -- Received: } from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 12 -- by } ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k05NJrdL005222 6, 12 } -- for {microscopy-at-microscopy.com} ; Thu, 5 Jan 2006 17:19:53 -0600 6, } 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- } Message-Id: {p06110400bfe35a0001a2-at-[206.69.208.22]} 6, 12 -- Date: } Thu, 5 Jan 2006 17:19:47 -0600 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: tsoumt-at-ilan.tpg.gov.tw (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: Canon pro1 onto trinocular port of Olympus } BX51 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
==============================Original Headers============================== 10, 25 -- From edelmare-at-muohio.edu Fri Jan 6 08:33:25 2006 10, 25 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 10, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06EXIIR006521 10, 25 -- for {microscopy-at-Microscopy.com} ; Fri, 6 Jan 2006 08:33:19 -0600 10, 25 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 10, 25 -- by mulnx12.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k06EWWHB001896; 10, 25 -- Fri, 6 Jan 2006 09:32:32 -0500 10, 25 -- Received: from emf03 ([134.53.14.97]) 10, 25 -- by mulnx24.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k06EWWSt016030; 10, 25 -- Fri, 6 Jan 2006 09:32:32 -0500 10, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 10, 25 -- To: tsoumt-at-ilan.tpg.gov.tw 10, 25 -- Date: Fri, 6 Jan 2006 09:33:29 -0500 10, 25 -- MIME-Version: 1.0 10, 25 -- Content-type: text/plain; charset=US-ASCII 10, 25 -- Content-transfer-encoding: 7BIT 10, 25 -- Subject: Re: [Microscopy] viaWWW: Canon pro1 onto trinocular port of Olympus BX51 10, 25 -- CC: microscopy-at-Microscopy.com 10, 25 -- Message-ID: {43BE3969.5908.ECA3945-at-localhost} 10, 25 -- Priority: normal 10, 25 -- In-reply-to: {200601060157.k061vebc022853-at-ns.microscopy.com} 10, 25 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 10, 25 -- X-Real-ConnectIP: 134.53.14.97 10, 25 -- X-Scanned-By: MIMEDefang 2.45 10, 25 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 ==============================End of - Headers==============================
Hello everybody, One of our EM lab users wants to buy triangular tungsten carbide knives for serial sectioning. Is there any advantage in buying that kind of knife? What is a life span (how many blocks/sections can be cut before it becomes dull)? Can they be resharpen? Are they good for EM resins (Epon, Spurrs ect.)? Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Fri Jan 6 11:00:05 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06H014x010343 1, 22 -- for {microscopy-at-microscopy.com} ; Fri, 6 Jan 2006 11:00:02 -0600 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1EuuwX-00070Q-01 1, 22 -- for {microscopy-at-microscopy.com} ; Fri, 06 Jan 2006 12:59:57 -0400 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 6 Jan 06 12:59:57 -0400 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 6 Jan 06 12:59:36 -0400 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Fri, 6 Jan 2006 12:10:46 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: LM tungsten carbide knives 1, 22 -- Message-ID: {43BE5E46.13266.E4BB6B-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
For glue to get section ribbons to stick together, I like to use Weldwood Contact Cement (avialable at your local hardware store) diluted 1:1 in xylene. I picked up this tip somewhere a few years ago, but don't remember where. Maybe it was from Microscopy!
I assume you have trimed the block to be a 4 sided pyramid with a flat top being the blockface from which sections will be cut. To apply glue, use a small end of a toothpick, dip in diluted glue, and dab onto one of the 4 sides of the block face, either top or bottom relative to orientation of block during sectioning. I usually tilt the block face so that the side I choose is nearly horizontal so the glue doesn't run down the block side, or over the actual face of the block. Be careful so that glue does not get onto the blockface itself. Also, work quickly as the glue will dry fast. Then reorient the block for sectioning.
There is also a product out there called Tacky Wax, available from EM supply companies, which you dab onto the side of the block face. You may or may not need to apply gentle heat near it to get it to spread evenly over the side of the blockface. Perhaps others may wish to comment on the use of Tacky Wax.
Hope this helps you!
Gib ---- Gib Ahlstrand Imaging Center University of Minbnesota St. Paul, MN 55108 ahlst007-at-umn.eud http://www.cbs.umn.edu/ic
rra-at-stowers-institute.org wrote: } Email: rra-at-stowers-institute.org } Name: Rhonda Allen } } Organization: Stowers Institute } } Title-Subject: [Filtered] glue to make ribbon's stick together } } Question: Hello, } I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions. } Thanks in adance. } Rhonda Allen } rra-at-stowers-institute.org
==============================Original Headers============================== 7, 20 -- From ahlst007-at-umn.edu Fri Jan 6 11:02:56 2006 7, 20 -- Received: from mtaout-m.tc.umn.edu (mtaout-m.tc.umn.edu [160.94.23.21]) 7, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06H2oln010411 7, 20 -- for {Microscopy-at-Microscopy.com} ; Fri, 6 Jan 2006 11:02:55 -0600 7, 20 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) by mtaout-m.tc.umn.edu with ESMTP; Fri, 6 Jan 2006 11:02:44 -0600 (CST) 7, 20 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 7, 20 -- Message-ID: {43BEA2AF.7030808-at-umn.edu} 7, 20 -- Date: Fri, 06 Jan 2006 11:02:39 -0600 7, 20 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 7, 20 -- Reply-To: ahlst007-at-umn.edu 7, 20 -- Organization: Imaging Center UM 7, 20 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 7, 20 -- X-Accept-Language: en-us, en 7, 20 -- MIME-Version: 1.0 7, 20 -- To: rra-at-stowers-institute.org, Microscopy-at-Microscopy.com 7, 20 -- Subject: Re: [Microscopy] viaWWW: glue to make ribbon's stick together 7, 20 -- References: {200601040322.k043ML3t002256-at-ns.microscopy.com} 7, 20 -- In-Reply-To: {200601040322.k043ML3t002256-at-ns.microscopy.com} 7, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Job Opening #B005608; Lab Technician/Engineer at IBM
There is an opening at the Lab Technician/Engineer level within the Materials Characterization and Analysis Group in the Research Division of IBM at the Almaden Research Center in San Jose, California. The job is a long term (three years) supplemental position with benefits.This position involves technical support in an advanced electron microscopy laboratory which emphasizes transmission electron microscopy. The technician works as a team member with professional (Ph.D.) microscopists and other scientists in a research laboratory. Duties involve primarily sample preparation methods including the conventional grinding, polishing, and ion milling techniques, focused ion beam (FIB ) method, as well as Microtome, etc. In addition, the candidate must be able to operate and perform routine maintenance on specimen preparation equipment, interact with customers (professional scientists), and prepare reports. The candidate must be able to work independently within set priorities, to keep abreast of new advances, and to interact smoothly within a team.
Experience with sample preparation and basic TEM imaging is a plus.
IBM is an equal opportunity employer. Women and Minorities are encouraged to apply. Information on the Almaden Research Center can be found at www.almaden.ibm.com
Interested parties may call or send resumes to me at the following address:
Dr. Philip M. Rice IBM Research Division Almaden Research Center 650 Harry Road, K19B/D1 tel: (408) 927-1442 fax: (408) 927-2100 email: pmrice-at-almaden.ibm.com
==============================Original Headers============================== 10, 26 -- From pmrice-at-almaden.ibm.com Fri Jan 6 11:23:44 2006 10, 26 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 10, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06HMhYt011005 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 6 Jan 2006 11:23:10 -0600 10, 26 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 10, 26 -- by e2.ny.us.ibm.com (8.12.11/8.12.11) with ESMTP id k06HMdHj029131 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 6 Jan 2006 12:22:39 -0500 10, 26 -- Received: from d01av01.pok.ibm.com (d01av01.pok.ibm.com [9.56.224.215]) 10, 26 -- by d01relay02.pok.ibm.com (8.12.10/NCO/VERS6.8) with ESMTP id k06HMd87122936 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 6 Jan 2006 12:22:39 -0500 10, 26 -- Received: from d01av01.pok.ibm.com (loopback [127.0.0.1]) 10, 26 -- by d01av01.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k06HMdBn004984 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 6 Jan 2006 12:22:39 -0500 10, 26 -- Received: from d01ml605.pok.ibm.com (d01ml605.pok.ibm.com [9.56.227.91]) 10, 26 -- by d01av01.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k06HMck9004981 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 6 Jan 2006 12:22:38 -0500 10, 26 -- Subject: TEM - Job Opening for TEM Specimen Preparation Lab Technician 10, 26 -- To: microscopy-at-microscopy.com 10, 26 -- X-Mailer: Lotus Notes Release 6.0.2CF1 June 9, 2003 10, 26 -- Message-ID: {OFBC79C14C.36D1DACC-ON882570EE.005F03ED-882570EE.005F72CD-at-us.ibm.com} 10, 26 -- From: Philip Rice {pmrice-at-almaden.ibm.com} 10, 26 -- Date: Fri, 6 Jan 2006 09:22:33 -0800 10, 26 -- X-MIMETrack: Serialize by Router on D01ML605/01/M/IBM(Release 7.0HF90 | November 16, 2005) at 10, 26 -- 01/06/2006 12:22:38 10, 26 -- MIME-Version: 1.0 10, 26 -- 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.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 for Electron Tomography 2006
Question: **************** First Announcement *************** The Fourth International Congress on Electron Tomography, (4ICET), will be held at Paradise Point, San Diego November 5-8, 2006.
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:
ď Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ď Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Univ. ofUltrect, Netherlands)
ď 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); TBA
ď Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ď Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ď 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.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 for Electron Tomography 2006
Question: **************** First Announcement *************** The Fourth International Congress on Electron Tomography, (4ICET), will be held at Paradise Point, San Diego November 5-8, 2006.
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:
ď Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ď Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Univ. ofUltrect, Netherlands)
ď 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); TBA
ď Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ď Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ď 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)
Year 10 was last year: I hope you can join us as the UBC Course enters its second decade.
Speaking for myself, and I am sure for our faculty, it is a source of great satisfaction to see how many of the contributors to this list are UBC Course Alumnae.
Thank you all.
Jim P.
Eleventh Annual INTERNATIONAL 12-Day Short Course on 3D Microscopy of Living Cells, June 10 - 22, 2006 (Pre-course: June 10)
Tenth, Post-course Workshop on 3D Image Processing, June 24 -26, 2006
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with the Departments of Pharmacology and Physiology and the Brain Research Centre, University of British Columbia, Vancouver, BC, Canada
DATES
Applications must be received by Tuesday, March 15, 2006 Deposit due Friday, April 15, 2006 Registration 5:00 - 7:00 PM Saturday, June 10, 2006 First Lecture 7:30 PM Saturday, June 10, 2006 Live-cell Course ends, noon Thursday, June 22, 2006 3D Image Processing Course, Saturday, June 24 - Monday, 26, 2006
APPLICATIONS DUE BY MARCH 15, 2006
APPLICATIONS Applicants must complete a questionnaire on the web to assess knowledge level, field of interest and proposed personal, live-cell, project. But don't let this put you off: if you plan to use 3D microscopy on living cells, we can usually find a way to make it work.
Enrollment will be limited to about 32 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms may be down-loaded from the WWW site at
http://www.3dcourse.ubc.ca/application.htm , or obtained from:
Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 250 N. Mills Street, Madison, WI, 53706 JBPAWLEY-at-WISC.EDU
Additional information is available from: http://www.3dcourse.ubc.ca/brochure.htm and links.
We expect to have at least 11, 3D microscope workstations for student use and there will be an international faculty of 20.
Application deadlines:
Application forms should be received for screening by March 15, 2006. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2006. In general, refunds of the deposit will only be possible if someone on the waiting list can take the place but this has not been a problem in previous years. The remaining balance is due before Registration.
Pre-Course Tuition (1/2 day Basic Optical principles) $125 (US) 3D Live-cell Course Tuition (includes lunches, snacks, 3 dinners, incl. the NEW Third Edition of the Handbook of Biological Confocal Microscopy): $2,750 (US) Workshop Tuition (includes lunches and snacks): $1,100 (US)
Room/board about $40/day (US) depending on room type.
I hope that this includes all of the information that you need, but if not, please get back to me.
Jim Pawley
-- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-262-9083 250 N. Mills St., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
==============================Original Headers============================== 27, 24 -- From jbpawley-at-wisc.edu Fri Jan 6 12:54:20 2006 27, 24 -- Received: from smtp4.wiscmail.wisc.edu (hermes.doit.wisc.edu [144.92.197.190]) 27, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06IrMoO013779 27, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 6 Jan 2006 12:53:44 -0600 27, 24 -- Received: from avs-daemon.smtp4.wiscmail.wisc.edu by smtp4.wiscmail.wisc.edu 27, 24 -- (iPlanet Messaging Server 5.2 HotFix 2.08 (built Sep 22 2005)) 27, 24 -- id {0ISO0090SPSXW6-at-smtp4.wiscmail.wisc.edu} for Microscopy-at-Microscopy.Com; 27, 24 -- Fri, 06 Jan 2006 12:53:21 -0600 (CST) 27, 24 -- Received: from [144.92.238.207] by smtp4.wiscmail.wisc.edu 27, 24 -- (iPlanet Messaging Server 5.2 HotFix 2.08 (built Sep 22 2005)) 27, 24 -- with ESMTPSA id {0ISO00LB4PSRMA-at-smtp4.wiscmail.wisc.edu} for 27, 24 -- Microscopy-at-Microscopy.Com; Fri, 06 Jan 2006 12:53:16 -0600 (CST) 27, 24 -- Date: Fri, 06 Jan 2006 12:53:14 -0600 27, 24 -- From: James Pawley {jbpawley-at-wisc.edu} 27, 24 -- Subject: First announcement: Eleventh International UBC Live-cell Course. 27, 24 -- X-Sender: jbpawley-at-wiscmail.wisc.edu 27, 24 -- To: "ListServer-at-MSA.Microscopy.Com" {Microscopy-at-Microscopy.Com} 27, 24 -- Message-id: {p06110415bfe46d18adc1-at-[144.92.238.207]} 27, 24 -- MIME-version: 1.0 27, 24 -- Content-type: text/plain; format=flowed; charset=us-ascii 27, 24 -- Content-transfer-encoding: 7BIT 27, 24 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=0.0.0.0 27, 24 -- X-Spam-PmxInfo: Server=avs-8, Version=5.1.1.222179, Antispam-Engine: 2.1.0.0, 27, 24 -- Antispam-Data: 2006.1.6.19, SenderIP=0.0.0.0 ==============================End of - Headers==============================
In our lab, there was a person using a thin coat of dental wax successfully on bottom side of a block. Melt dental in a beaker and apply a tiny bit to a shaped block with a warm spatula. You may have to trim out excess wax.
I have Tackiwax also. It came in a tiny bottle. It seems to me that it is repackaged dental wax without color. On the other hand, I may be wrong this material.
Ann Fook Yang EM Unit/ Unite EM AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 yanga-at-agr.gc.ca Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: ahlst007-at-umn.edu [mailto:ahlst007-at-umn.edu] Sent: Friday, January 06, 2006 5:35 PM To: Yang, Ann-Fook
Rhonda,
For glue to get section ribbons to stick together, I like to use Weldwood Contact Cement (avialable at your local hardware store) diluted 1:1 in xylene. I picked up this tip somewhere a few years ago, but don't remember where. Maybe it was from Microscopy!
I assume you have trimed the block to be a 4 sided pyramid with a flat top being the blockface from which sections will be cut. To apply glue, use a small end of a toothpick, dip in diluted glue, and dab onto one of the 4 sides of the block face, either top or bottom relative to orientation of block during sectioning. I usually tilt the block face so that the side I choose is nearly horizontal so the glue doesn't run down the block side, or over the actual face of the block. Be careful so that glue does not get onto the blockface itself. Also, work quickly as the glue will dry fast. Then reorient the block for sectioning.
There is also a product out there called Tacky Wax, available from EM supply companies, which you dab onto the side of the block face. You may or may not need to apply gentle heat near it to get it to spread evenly over the side of the blockface. Perhaps others may wish to comment on the use of Tacky Wax.
Hope this helps you!
Gib ---- Gib Ahlstrand Imaging Center University of Minbnesota St. Paul, MN 55108 ahlst007-at-umn.eud http://www.cbs.umn.edu/ic
rra-at-stowers-institute.org wrote: } Email: rra-at-stowers-institute.org } Name: Rhonda Allen } } Organization: Stowers Institute } } Title-Subject: [Filtered] glue to make ribbon's stick together } } Question: Hello, } I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions. } Thanks in adance. } Rhonda Allen } rra-at-stowers-institute.org
==============================Original Headers============================== 7, 20 -- From ahlst007-at-umn.edu Fri Jan 6 11:02:56 2006 7, 20 -- Received: from mtaout-m.tc.umn.edu (mtaout-m.tc.umn.edu [160.94.23.21]) 7, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06H2oln010411 7, 20 -- for {Microscopy-at-Microscopy.com} ; Fri, 6 Jan 2006 11:02:55 -0600 7, 20 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) by mtaout-m.tc.umn.edu with ESMTP; Fri, 6 Jan 2006 11:02:44 -0600 (CST) 7, 20 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 7, 20 -- Message-ID: {43BEA2AF.7030808-at-umn.edu} 7, 20 -- Date: Fri, 06 Jan 2006 11:02:39 -0600 7, 20 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 7, 20 -- Reply-To: ahlst007-at-umn.edu 7, 20 -- Organization: Imaging Center UM 7, 20 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 7, 20 -- X-Accept-Language: en-us, en 7, 20 -- MIME-Version: 1.0 7, 20 -- To: rra-at-stowers-institute.org, Microscopy-at-Microscopy.com 7, 20 -- Subject: Re: [Microscopy] viaWWW: glue to make ribbon's stick together 7, 20 -- References: {200601040322.k043ML3t002256-at-ns.microscopy.com} 7, 20 -- In-Reply-To: {200601040322.k043ML3t002256-at-ns.microscopy.com} 7, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 16, 29 -- From YANGA-at-AGR.GC.CA Mon Jan 9 09:25:19 2006 16, 29 -- Received: from agroutsmtp1.agr.gc.ca (agroutsmtp1.agr.gc.ca [192.197.71.175]) 16, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k09FPHkv029897 16, 29 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 09:25:18 -0600 16, 29 -- Received: from agrin1-old.agr.gc.ca (agrgate.agr.ca [192.197.71.189]) 16, 29 -- by agroutsmtp1.agr.gc.ca (8.12.8/8.12.8) with ESMTP id k09FPDHp017974 16, 29 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 10:25:13 -0500 16, 29 -- Received: from onncrxcn1.AGR.GC.CA ([10.117.15.130]) 16, 29 -- by agrin1-old.agr.gc.ca (8.12.8/8.12.8) with ESMTP id k09FQiUY023252 16, 29 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 10:26:45 -0500 16, 29 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn1.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 16, 29 -- Mon, 9 Jan 2006 10:25:08 -0500 16, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 16, 29 -- content-class: urn:content-classes:message 16, 29 -- MIME-Version: 1.0 16, 29 -- Content-Type: text/plain; 16, 29 -- charset="iso-8859-1" 16, 29 -- Subject: RE: [Microscopy] glue to make ribbon's stick together 16, 29 -- Date: Mon, 9 Jan 2006 10:25:06 -0500 16, 29 -- Message-ID: {E035A9C87303AE4AB9BA10FD8324DFB10243345A-at-onncrxms3.agr.gc.ca} 16, 29 -- X-MS-Has-Attach: 16, 29 -- X-MS-TNEF-Correlator: 16, 29 -- Thread-Topic: [Microscopy] glue to make ribbon's stick together 16, 29 -- Thread-Index: AcYT2iOC0AiPsPOzQuGBu5lpBYTxtgBUcMiw 16, 29 -- From: "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 16, 29 -- To: {microscopy-at-microscopy.com} 16, 29 -- X-OriginalArrivalTime: 09 Jan 2006 15:25:08.0115 (UTC) FILETIME=[DFA16E30:01C61530] 16, 29 -- Content-Transfer-Encoding: 8bit 16, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k09FPHkv029897 ==============================End of - Headers==============================
Does anyone know if the various glues suggested vaporise in the TEM? If so does this cause any problems?
Dave
-----Original Message----- X-from: YANGA-at-AGR.GC.CA [mailto:YANGA-at-AGR.GC.CA] Sent: 09 January 2006 15:28 To: David Patton
In our lab, there was a person using a thin coat of dental wax successfully on bottom side of a block. Melt dental in a beaker and apply a tiny bit to a shaped block with a warm spatula. You may have to trim out excess wax.
I have Tackiwax also. It came in a tiny bottle. It seems to me that it is repackaged dental wax without color. On the other hand, I may be wrong this material.
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: ahlst007-at-umn.edu [mailto:ahlst007-at-umn.edu] Sent: Friday, January 06, 2006 5:35 PM To: Yang, Ann-Fook
Rhonda,
For glue to get section ribbons to stick together, I like to use Weldwood Contact Cement (avialable at your local hardware store) diluted 1:1 in xylene. I picked up this tip somewhere a few years ago, but don't remember where. Maybe it was from Microscopy!
I assume you have trimed the block to be a 4 sided pyramid with a flat top being the blockface from which sections will be cut. To apply glue, use a small end of a toothpick, dip in diluted glue, and dab onto one of the 4 sides of the block face, either top or bottom relative to orientation of block during sectioning. I usually tilt the block face so that the side I choose is nearly horizontal so the glue doesn't run down the block side, or over the actual face of the block. Be careful so that glue does not get onto the blockface itself. Also, work quickly as the glue will dry fast. Then reorient the block for sectioning.
There is also a product out there called Tacky Wax, available from EM supply companies, which you dab onto the side of the block face. You may or may not need to apply gentle heat near it to get it to spread evenly over the side of the blockface. Perhaps others may wish to comment on the use of Tacky Wax.
Hope this helps you!
Gib ---- Gib Ahlstrand Imaging Center University of Minbnesota St. Paul, MN 55108 ahlst007-at-umn.eud http://www.cbs.umn.edu/ic
rra-at-stowers-institute.org wrote: } Email: rra-at-stowers-institute.org } Name: Rhonda Allen } } Organization: Stowers Institute } } Title-Subject: [Filtered] glue to make ribbon's stick together } } Question: Hello, } I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions. } Thanks in adance. } Rhonda Allen } rra-at-stowers-institute.org
==============================Original Headers============================== 7, 20 -- From ahlst007-at-umn.edu Fri Jan 6 11:02:56 2006 7, 20 -- Received: from mtaout-m.tc.umn.edu (mtaout-m.tc.umn.edu [160.94.23.21]) 7, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k06H2oln010411 7, 20 -- for {Microscopy-at-Microscopy.com} ; Fri, 6 Jan 2006 11:02:55 -0600 7, 20 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) by mtaout-m.tc.umn.edu with ESMTP; Fri, 6 Jan 2006 11:02:44 -0600 (CST) 7, 20 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 7, 20 -- Message-ID: {43BEA2AF.7030808-at-umn.edu} 7, 20 -- Date: Fri, 06 Jan 2006 11:02:39 -0600 7, 20 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 7, 20 -- Reply-To: ahlst007-at-umn.edu 7, 20 -- Organization: Imaging Center UM 7, 20 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 7, 20 -- X-Accept-Language: en-us, en 7, 20 -- MIME-Version: 1.0 7, 20 -- To: rra-at-stowers-institute.org, Microscopy-at-Microscopy.com 7, 20 -- Subject: Re: [Microscopy] viaWWW: glue to make ribbon's stick together 7, 20 -- References: {200601040322.k043ML3t002256-at-ns.microscopy.com} 7, 20 -- In-Reply-To: {200601040322.k043ML3t002256-at-ns.microscopy.com} 7, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 16, 29 -- From YANGA-at-AGR.GC.CA Mon Jan 9 09:25:19 2006 16, 29 -- Received: from agroutsmtp1.agr.gc.ca (agroutsmtp1.agr.gc.ca [192.197.71.175]) 16, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k09FPHkv029897 16, 29 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 09:25:18 -0600 16, 29 -- Received: from agrin1-old.agr.gc.ca (agrgate.agr.ca [192.197.71.189]) 16, 29 -- by agroutsmtp1.agr.gc.ca (8.12.8/8.12.8) with ESMTP id k09FPDHp017974 16, 29 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 10:25:13 -0500 16, 29 -- Received: from onncrxcn1.AGR.GC.CA ([10.117.15.130]) 16, 29 -- by agrin1-old.agr.gc.ca (8.12.8/8.12.8) with ESMTP id k09FQiUY023252 16, 29 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 10:26:45 -0500 16, 29 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn1.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 16, 29 -- Mon, 9 Jan 2006 10:25:08 -0500 16, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 16, 29 -- content-class: urn:content-classes:message 16, 29 -- MIME-Version: 1.0 16, 29 -- Content-Type: text/plain; 16, 29 -- charset="iso-8859-1" 16, 29 -- Subject: RE: [Microscopy] glue to make ribbon's stick together 16, 29 -- Date: Mon, 9 Jan 2006 10:25:06 -0500 16, 29 -- Message-ID: {E035A9C87303AE4AB9BA10FD8324DFB10243345A-at-onncrxms3.agr.gc.ca} 16, 29 -- X-MS-Has-Attach: 16, 29 -- X-MS-TNEF-Correlator: 16, 29 -- Thread-Topic: [Microscopy] glue to make ribbon's stick together 16, 29 -- Thread-Index: AcYT2iOC0AiPsPOzQuGBu5lpBYTxtgBUcMiw 16, 29 -- From: "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 16, 29 -- To: {microscopy-at-microscopy.com} 16, 29 -- X-OriginalArrivalTime: 09 Jan 2006 15:25:08.0115 (UTC) FILETIME=[DFA16E30:01C61530] 16, 29 -- Content-Transfer-Encoding: 8bit 16, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k09FPHkv029897 ==============================End of - Headers==============================
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==============================Original Headers============================== 29, 33 -- From David.Patton-at-uwe.ac.uk Mon Jan 9 09:32:45 2006 29, 33 -- Received: from mailapp02.uwe.ac.uk (mailapp02.uwe.ac.uk [164.11.132.63]) 29, 33 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k09FWhRa002940 29, 33 -- for {Microscopy-at-Microscopy.com} ; Mon, 9 Jan 2006 09:32:44 -0600 29, 33 -- Received: from (164.11.132.62) by mailapp02.uwe.ac.uk via smtp 29, 33 -- id 0461_8549066c_8125_11da_8b14_0002b3c90020; 29, 33 -- Mon, 09 Jan 2006 15:35:11 +0000 29, 33 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 29, 33 -- (egen-uwe01.campus.ads.uwe.ac.uk [164.11.249.121]) 29, 33 -- by mta02.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 29, 33 -- 2005)) with ESMTP id {0ISU00H0T0IHDB-at-mta02.uwe.ac.uk} for 29, 33 -- Microscopy-at-Microscopy.com; Mon, 09 Jan 2006 15:32:41 +0000 (GMT) 29, 33 -- Date: Mon, 09 Jan 2006 15:32:40 +0000 29, 33 -- From: David Patton {David.Patton-at-uwe.ac.uk} 29, 33 -- Subject: RE: [Microscopy] RE: glue to make ribbon's stick together 29, 33 -- To: Microscopy-at-Microscopy.com 29, 33 -- Message-id: {F247F674896BE243AD8263C5280E2BDBF8507D-at-egen-uwe01} 29, 33 -- MIME-version: 1.0 29, 33 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.6944.0 29, 33 -- Content-type: text/plain; 29, 33 -- charset="utf-8" 29, 33 -- Content-class: urn:content-classes:message 29, 33 -- Thread-topic: [Microscopy] RE: glue to make ribbon's stick together 29, 33 -- Thread-index: AcYVMVNhlhbSoF3kQxeNI9avIx8RPAAAF3QQ 29, 33 -- X-MS-Has-Attach: 29, 33 -- X-MS-TNEF-Correlator: 29, 33 -- X-NAI-Spam-Score: -0.3 29, 33 -- X-NAI-Spam-Rules: 1 Rules triggered 29, 33 -- BAYES_30=-0.3 29, 33 -- X-NAIMIME-Disclaimer: 1 29, 33 -- X-NAIMIME-Modified: 1 29, 33 -- Content-Transfer-Encoding: 8bit 29, 33 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k09FWhRa002940 ==============================End of - Headers==============================
I posted this question a couple of weeks ago, but probably the timing was wrong (just before the holidays), no answers at all. So I am posting again:
I would need a tip on a good (commercially available) antibody against His tag for EM - Tokuyasu technique. It should tolerate at least light (0.1-0.2%) glutaraldehyde fixation. A rabbit polyclonal would be preferable, but a working monoclonal would do as well.
Another unrelated question - I would need to label macrophages in sections (again, Tokuyasu). Any idea for a good macrophage marker?
Thanks,
Michal
==============================Original Headers============================== 7, 19 -- From M_Jarnik-at-fccc.edu Mon Jan 9 10:03:32 2006 7, 19 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 7, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k09G3VhJ016139 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 10:03:32 -0600 7, 19 -- Received: from fccc.edu (emf4.fccc.edu [131.249.9.170]) 7, 19 -- by azah.fccc.edu (8.12.11/8.12.11) with ESMTP id k09G3USd021508 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 9 Jan 2006 11:03:30 -0500 (EST) 7, 19 -- Message-ID: {43C28952.4040809-at-fccc.edu} 7, 19 -- Date: Mon, 09 Jan 2006 11:03:30 -0500 7, 19 -- From: Michal Jarnik {M_Jarnik-at-fccc.edu} 7, 19 -- Reply-To: M_Jarnik-at-fccc.edu 7, 19 -- Organization: Fox Chase Cancer Center 7, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 7, 19 -- X-Accept-Language: en,cs 7, 19 -- MIME-Version: 1.0 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- Subject: His Tag antibody for EM 7, 19 -- Content-Type: text/plain; charset=us-ascii; format=flowed 7, 19 -- Content-Transfer-Encoding: 7bit ==============================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 mjo10-at-psu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mjo10-at-psu.edu Name: Matthew Olszta
Organization: Penn State University
Title-Subject: [Filtered] Vanadium oxide thin films
Question: I wanted to know if anyone has had any experience with TEM sample preparation and analysis of vanadium oxide thin films. The reason I ask is that certain vanadium oxide crystal structures (VO2 and V2O5) are soluble or slightly soluble in water, but using water in polishing I have not yet seen any appreciable dissolution in my cross-sections.
If others have worked with these materials, or other water sensitive materials, do you have any suggestions or comments on sample preparation in order to avoid artifacts?
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 mjo10-at-psu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mjo10-at-psu.edu Name: Matthew Olszta
Organization: Penn State University
Title-Subject: [Filtered] Vanadium oxide thin films
Question: I wanted to know if anyone has had any experience with TEM sample preparation and analysis of vanadium oxide thin films. The reason I ask is that certain vanadium oxide crystal structures (VO2 and V2O5) are soluble or slightly soluble in water, but using water in polishing I have not yet seen any appreciable dissolution in my cross-sections.
If others have worked with these materials, or other water sensitive materials, do you have any suggestions or comments on sample preparation in order to avoid artifacts?
I am not sure about vaporization issues, but I have used another glue. Using a sharpened toothpick dipped in acetone, I have dissolved glue from 3M adhesive tape ("Magic Tape") to apply thin amounts of glue to the top surface of the block when trying to obtain serial sections. I have not noted any contamination problems, but clearly you want to refrain from getting any of this solvent on the cutting face.
However, when collecting serial sections, glue should only be your last resort. When properly trimmed with a fresh razor blade (I like the chromium steel blades from "Wilkinson Sword Blades") so that both top edge and bottom edge are parallel to each other and to the diamond knife edge, most Epon-like resins allow serial sections to stick together quite well. I don't know how well this applies to Spurr's in particular, but I expect the same results. The cleaner those edges are, and the more parallel they are to each other and to the diamond, the better the result. Acrylic resins like LR White or LR Gold tend to be much more brittle, and those would be more difficult to handle in series.
Dave Hall
For a more exhaustive discussion of serial sectioning, see Hall (1995) Methods in Cell Biology, C. elegans: Modern Biological Analysis of an Organism. Epstein and Shakes (eds) Academic Press. pp. 395-436. -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
==============================Original Headers============================== 6, 21 -- From hall-at-aecom.yu.edu Tue Jan 10 10:06:01 2006 6, 21 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 6, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0AG61X7031584 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 10:06:01 -0600 6, 21 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 6, 21 -- by mailgw.aecom.yu.edu (8.12.11/8.12.11) with SMTP id k0AG60vS020160 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 11:06:00 -0500 6, 21 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 6, 21 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006011011062608439 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 11:06:26 -0500 6, 21 -- Received: from [129.98.90.160] (worm.aecom.yu.edu [129.98.90.160]) 6, 21 -- by post.aecom.yu.edu (Postfix) with ESMTP id 340A12FC0 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 11:06:00 -0500 (EST) 6, 21 -- Mime-Version: 1.0 6, 21 -- X-Sender: hall-at-mailserver.aecom.yu.edu 6, 21 -- Message-Id: {a05100316bfe98b41a356-at-[129.98.90.160]} 6, 21 -- Date: Tue, 10 Jan 2006 11:05:56 -0500 6, 21 -- To: microscopy-at-microscopy.com 6, 21 -- From: David Hall {hall-at-aecom.yu.edu} 6, 21 -- Subject: glue to make ribbon's stick together 6, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Matt, I don't have experience to work with vanadium oxide, but some superconductive materials which were not working well with water. If you use water only for your grinding procedure for your case, now you can try dry-grinding, at least for the final stage. or you can try leaving thicker for the final ion mill (like 45 micron) if it is not hard to be ion-milled under Ar+ ion beam, but this may cause some beam damage or contamination if there is not cold stage and the milling time is too long. again to try higher rate for the early milling stage and to reduce it at the final may help with this. or you could try methanol or ethanol for your grinding if the "wet" procedure is necessary. also you can try FIB. it is dry, but may consider its beam damage if your material is sensitive to this. Hope it can help
Sincerely Long Li _________________________________________________________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
----- Original Message ----- X-from: {mjo10-at-psu.edu} To: {Longli_tem-at-hotmail.com} Sent: Monday, January 09, 2006 7:39 PM
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Email: mkamrath-at-nalco.com Name: Mike Kamrath
Organization: Nalco Co., Naperville IL
Title-Subject: [Filtered] Colloidal Silica Particle Size Distribution by TEM
Question: I was looking for help with a problem we have related to particle size distributions (PSD) with colloidal silicas as measured by TEM. We normally dilute a colloidal silica solution in an aqueous solution containing a few drops of nonionic surfactant to help disperse the particles (5-100 nm). After placing a drop on a copper grid (Formvar) we dry it at 100 oC for 10 minutes and place in the TEM for PSD. We are trying to automate the process with counting software but the software has problems identifying individual particles when they are touching one another. I have two questions related to this: 1) Does anyone know of software capable of easily separating touching particles and counting individually? We are currently using Image-Pro Plus and I am aware of free software from NIH (Scion) which seems to have similar problems. 2) Does anybody have suggestions for a better solvent system or surfactant that might separate the particles better? Or is the drying process forcing the particles together and would a vacuum drying or other method work better?
A couple of months ago, I repumped in situ my chronically leaky PGT Be-window EDS detector, and ran 50 deg C air into the dewar while re-evacuating using the chamber vacuum via a hose from the sample introduction port of the JEOL 840 on which the detector is mounted.
The re-evacuation worked well, in that the LN2 consumption is now back to normal, but after recooling and switching on the bias again, there was no appreciable low-energy response -- no visible Cu L peak at all!
I realised that grease from the O-ring sliding seal through which the tube of the detector enters the chamber had melted and run down onto the Be window. Cleaning of the window with Freon restored the Cu L intensity, and all was well, until it became obvious that the window was re-oiling again at a much faster rate than ever before.
Because we do quantitative mineral analysis, I keep a good handle on the window condition, evidenced by both Cu L and Na K responses. I have been using this detector for about five years and not found it neccesary to clean the window before, but now the Na response halves in a few weeks. Recleaning with Freon restores the response again.
In an effort to fix this, I have:-
- changed the rotary pump oil - installed an alumina-pellet foreline trap - changed the DP oil (Santovac, but the old charge was still light-colored)
but still the darned window is oiling up.
I'm running out of ideas. It seems that either the chamber atmosphere has become markedly oilier than before, or the detector window is now running colder than before.
My money is on the latter, as it seems too much of a coincidence for some change in the back-streaming performance of the 840 to have occurred at the same time as the repumping of the detector, but I can't see what might have happened.
Anyone got any suggestions?
Happy New Year
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 16, 26 -- From r.sims-at-auckland.ac.nz Tue Jan 10 19:00:56 2006 16, 26 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) 16, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0B10td7001103 16, 26 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 19:00:56 -0600 16, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 16, 26 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id F369034958 16, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 14:00:53 +1300 (NZDT) 16, 26 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) 16, 26 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 16, 26 -- with ESMTP id 31308-14 for {microscopy-at-microscopy.com} ; 16, 26 -- Wed, 11 Jan 2006 14:00:46 +1300 (NZDT) 16, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 16, 26 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 29EF235637 16, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 13:59:05 +1300 (NZDT) 16, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 16, 26 -- To: microscopy-at-microscopy.com 16, 26 -- Date: Wed, 11 Jan 2006 13:59:04 +1300 16, 26 -- MIME-Version: 1.0 16, 26 -- Subject: Oil on detector window 16, 26 -- Message-ID: {43C50F28.11970.13D9DFF-at-localhost} 16, 26 -- Priority: normal 16, 26 -- X-mailer: Pegasus Mail for Windows (4.21c) 16, 26 -- Content-type: text/plain; charset=US-ASCII 16, 26 -- Content-transfer-encoding: 7BIT 16, 26 -- Content-description: Mail message body 16, 26 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
First, may I ask what you have done to ensure that your support films are hydrophilic? Carbon coated polyvinylformal supports become increasingly hydrophobic with age. We get good results by treating with spectro-grade acetone for 30 min prior to use. We have also used a short plasma-ash in air to improve wettability. Such treatments improves dispersion and reduces drying artifacts.
We use the analySIS Five software from Soft Imaging System for image analysis. This program has a useful separator based upon a watershed algorithm. I have also written a custom watershed module for some applications. We also use a guard frame to eliminate bias from the higher probability that larger particles touch the image boundaries compared to smaller particles. We prefer this approach to the Miles-Lantejoul algorithm. Using an extension to the analySIS Automater we can queue up images from several samples from any of our microscopes and let them run unattended.
We have compared results obtained by simply deleting agglomerates with the results obtained by separation. Even if we use a separator, we typically measure the maximum intrusion distance [this is a custom module we wrote] for each particle and flag those which are not convex as agglomerates. We always keep track of the area fraction of agglomerates and reject analyses where this is too high. Usually we can find preparation conditions to keep the area fraction of agglomerates below 10%.
One of the strengths of the analySIS program is that the macro language is almost a full subset of C and if one needs a computationally-intensive function (like the maximum intrusion distance) it is easy to import a C function from a custom DLL compiled with a good optimizing compiler, like Visual C++.
Disclaimer: I have no financial interest in Soft Imaging System, writing only as a satisfied user.
John R. Minter Eastman Kodak Co. Research Laboratories Foundation Science and Technology Center john.r.minter-at-kodak.com (work) jrminter-at-rochester.rr.com (home)
-----Original Message----- X-from: mkamrath-at-nalco.com [mailto:mkamrath-at-nalco.com] Sent: Tuesday, January 10, 2006 7:28 PM To: jrminter-at-rochester.rr.com
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please copy both mkamrath-at-nalco.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mkamrath-at-nalco.com Name: Mike Kamrath
Organization: Nalco Co., Naperville IL
Title-Subject: [Filtered] Colloidal Silica Particle Size Distribution by TEM
Question: I was looking for help with a problem we have related to particle size distributions (PSD) with colloidal silicas as measured by TEM. We normally dilute a colloidal silica solution in an aqueous solution containing a few drops of nonionic surfactant to help disperse the particles (5-100 nm). After placing a drop on a copper grid (Formvar) we dry it at 100 oC for 10 minutes and place in the TEM for PSD. We are trying to automate the process with counting software but the software has problems identifying individual particles when they are touching one another. I have two questions related to this: 1) Does anyone know of software capable of easily separating touching particles and counting individually? We are currently using Image-Pro Plus and I am aware of free software from NIH (Scion) which seems to have similar problems. 2) Does anybody have suggestions for a better solvent system or surfactant that might separate the particles better? Or is the drying p! rocess forcing the particles together and would a vacuum drying or other method work better?
I am looking for a simple probe-type sonicator that will be used to make holey films.
We have used an old one in the past that is no longer available. It is used to aerate a formvar/glycerol solution sufficiently to make very small bubbles. When a slide is dipped into the solution, the resulting film has holes of a size dependent on the length of time used for the sonication.
Any recommendations welcomed but hopefully I can find one on the less expensive side.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 6, 21 -- From dsherman-at-purdue.edu Wed Jan 11 08:11:13 2006 6, 21 -- Received: from exchange.purdue.edu (1061exfe03.adpc.purdue.edu [128.210.63.225]) 6, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0BEBDF0028321 6, 21 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 08:11:13 -0600 6, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.235]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Wed, 11 Jan 2006 09:10:23 -0500 6, 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 ; 6, 21 -- Wed, 11 Jan 2006 14:10:15 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 6, 21 -- Date: Wed, 11 Jan 2006 09:11:05 -0500 6, 21 -- Subject: Sonicator 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {BFEA7C29.C52D%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Sonicator 6, 21 -- Thread-Index: AcYWuNwnGsctLIKsEdqMVgARJN08Mg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 11 Jan 2006 14:10:23.0149 (UTC) FILETIME=[C3356DD0:01C616B8] ==============================End of - Headers==============================
sue- For what it's worth, and to get in synch with anyone out there trying tests such as i described, last week I repeated my 40 year old MnO4 tests in water on all the liquid resin components, adding 1-2 small drops to 2.5 ml dilute transparent purple MnO4 sol'n (2.5 ml water, 2 drops 2% KMnO4). I found that NMA instantly turns the sol'n to clear brown (guess I'd misremembered that it made a precipitate before), and DMP-30 does so almost as fast. All the other components, including Araldite 506, do the same thing over periods of 5-200 minutes, Araldite 506 clearly slower than Epon 812. Some make a precipitate, some don't. By next day, nothing retains any purple, all test sol'ns are brown. So the lesson is, MnO4 can act eventually destructively on any sections or their components, given time enough. -mike-
} hi mike...thanks for all your help!!! } sue
-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
I am looking into acquiring a new field emission SEM which will be used mainly for failure analysis of metals, but also plastics, paper, an occasional drop of grease, and anything else that wanders through the lab (but rarely biological samples). The field emission SEMs that I am considering are the JEOL JSM-7000F, the ZEISS Supra 40 (VP), the Hitachi S-4300SE/N, and the FEI Quanta 600 FEG. For microanalysis systems, I am considering the newest versions from Thermoelectron, Oxford and EDAX. I welcome any comments that you can offer as to the function (or malfunction) of the above instruments and anaylsis systems, as well as possible electro-magnetic interference issues, user interface issues, interface problems between the SEM and microanalysis systems, and the reliability/response time of the service representatives from the above companies.
Thank you very much for your time!
Sincerely,
Robin Moresi General Dynamics - Advanced Information Systems 100 Plastics Ave. Pittsfield, MA 01201
Tel. (413) 662-2889
==============================Original Headers============================== 8, 26 -- From Robin.Moresi-at-gd-ais.com Wed Jan 11 11:21:00 2006 8, 26 -- Received: from mnblin01.mnb.gd-ais.com (mnblin01.mnb.gd-ais.com [206.11.149.28]) 8, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0BHKxPT012197 8, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 11:20:59 -0600 8, 26 -- Received: from mnbm01-fes01.ad.gd-ais.com (mnbm01-fes01.mnb.gd-ais.com [160.207.224.15]) 8, 26 -- by mnblin01.mnb.gd-ais.com (8.12.11/8.12.11) with SMTP id k0BHMC0b006944 8, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 11:22:12 -0600 8, 26 -- Received: from MAPF01-MAIL01.ad.gd-ais.com ([166.16.220.104]) by mnbm01-fes01.ad.gd-ais.com with Microsoft SMTPSVC(6.0.3790.1830); 8, 26 -- Wed, 11 Jan 2006 11:21:06 -0600 8, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 26 -- Content-class: urn:content-classes:message 8, 26 -- MIME-Version: 1.0 8, 26 -- Content-Type: text/plain; 8, 26 -- charset="us-ascii" 8, 26 -- Subject: Field Emission SEM question 8, 26 -- Date: Wed, 11 Jan 2006 12:20:47 -0500 8, 26 -- Message-ID: {5DC7DC21AC968A46AEF8265C6D623C15957A70-at-MAPF01-MAIL01.ad.gd-ais.com} 8, 26 -- X-MS-Has-Attach: 8, 26 -- X-MS-TNEF-Correlator: 8, 26 -- Thread-Topic: Field Emission SEM question 8, 26 -- Thread-Index: AcYWzfPqVEoK4BtUSPGMgcimUwgWZgAADOoAAAEzsbA= 8, 26 -- From: "Moresi, Robin T." {Robin.Moresi-at-gd-ais.com} 8, 26 -- To: {microscopy-at-microscopy.com} 8, 26 -- X-OriginalArrivalTime: 11 Jan 2006 17:21:06.0964 (UTC) FILETIME=[68410540:01C616D3] 8, 26 -- Content-Transfer-Encoding: 8bit 8, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0BHKxPT012197 ==============================End of - Headers==============================
I am looking for a manual for a SORVALL MT2-B ultra microtome. I will be happy to pay for copying and shipping costs, but would greatly appreciate if someone would be willing to copy their manual for me.
Thanks, Elke
Elke Buschbeck Biological Sciences University of Cincinnati Cincinnati, OH elke.buschbeck-at-uc.edu
==============================Original Headers============================== 6, 17 -- From elke.buschbeck-at-uc.edu Wed Jan 11 13:22:43 2006 6, 17 -- Received: from ms-smtp-02-eri0.ohiordc.rr.com (ms-smtp-02-smtplb.ohiordc.rr.com [65.24.5.136]) 6, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0BJMeEv026815 6, 17 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 13:22:42 -0600 6, 17 -- Received: from Mainbrain.uc.edu (cpe-65-27-235-87.cinci.res.rr.com [65.27.235.87]) 6, 17 -- by ms-smtp-02-eri0.ohiordc.rr.com (8.12.10/8.12.7) with ESMTP id k0BJMZg6002796 6, 17 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 14:22:37 -0500 (EST) 6, 17 -- Message-Id: {6.1.2.0.2.20060111141640.08deb160-at-ucmail8.uc.edu} 6, 17 -- X-Sender: buschbek-at-ucmail8.uc.edu 6, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.2.0 6, 17 -- Date: Wed, 11 Jan 2006 14:22:35 -0500 6, 17 -- To: Microscopy-at-microscopy.com 6, 17 -- From: Elke Buschbeck {elke.buschbeck-at-uc.edu} 6, 17 -- Subject: Need manual for Sorvall MT2-B ultra microtome 6, 17 -- Mime-Version: 1.0 6, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 17 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
What has happened to you has happened to many before you!
Once you reach the point of contaminating your window with RP fluid it will have contaminated the complete vacuum system. Two solutions. EITHER take everything apart and clean it, yes ALL the vacuum lines and column liners. OR use a hair drier to bake the pumping lines to drive off the contamination as best as possible, as well as cleaning the parts you are easily able to reach.
Many times in my service technician career I have been forced to take this route. It puts you out of action for a day or so whilst the system recovers but in the main it seems to be a cure. BUT if you do not tackle the fore line trap idea it will happen all over again - our chemical test said it was RP fluid by the way.
Good luck
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: {r.sims-at-auckland.ac.nz} To: {protrain-at-emcourses.com} Sent: Wednesday, January 11, 2006 1:02 AM
Will the person that wanted the MT2-B manual please contact me. I found the one I was looking for after I deleted your post.
Mannie Steglich msteglic-at-mdanderson.org
==============================Original Headers============================== 2, 15 -- From msteglic-at-mdanderson.org Wed Jan 11 15:43:11 2006 2, 15 -- Received: from UTM-MAIL04A.mdacc.tmc.edu (utmlnmail04nt.mdacc.tmc.edu [143.111.84.152]) 2, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0BLhB5n014618 2, 15 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Jan 2006 15:43:11 -0600 2, 15 -- To: Microscopy-at-MSA.Microscopy.Com 2, 15 -- Subject: MT2-B Manual 2, 15 -- MIME-Version: 1.0 2, 15 -- X-Mailer: Lotus Notes Release 5.0.11 July 24, 2002 2, 15 -- Message-ID: {OF72D353B1.75A3682D-ON862570F3.007713B0-862570F3.00774DE6-at-mdacc.tmc.edu} 2, 15 -- From: msteglic-at-mdanderson.org 2, 15 -- Date: Wed, 11 Jan 2006 15:42:54 -0600 2, 15 -- X-MIMETrack: Serialize by Router on UTM-MAIL04A/HOU/UTMDACC(Release 5.0.11 |July 24, 2002) at 2, 15 -- 01/11/2006 03:42:58 PM, 2, 15 -- Serialize complete at 01/11/2006 03:42:58 PM 2, 15 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Many thanks for the replies and suggestions so far, however, please note that this is a new condition.
For the past five years the oiling has not been a problem, and it has started to occur only since the repumping operation described below in tedious detail.
There have been no changes to the system apart from the three steps taken, one at a time, in an attempt to fix the problem (changing the RP oil; installing the foreline trap; and changing the DP oil), and they have made no difference.
I think it would be too much of a coincidence for the cause of the oiling to be unrelated to the repumping operation, so I think that complex cures such as installing LN2 traps, plasma cleaning, complete stripping and cleaning, etc, do not address the question, which is "what has changed to accelerate the detector window oiling so greatly, and how can I get back to the previous state?"
I had thought that perhaps a drop of melted grease might have dripped into the DP, and be decomposing in the Santovac, which propelled me to change the Santovac, but I now realise that this is very unlikely to be the case, as the DP is not directly beneath the sample chamber, and even if it were, such a drop of molten grease would not make it through the water-cooled baffle above the DP. Plus, of course, the fact that very thoroughly cleaning both of the DPs and both of their water-cooled baffles has made no difference at all to the problem.
Incidentally, the Santovac, after five years' continuous operation, was only a pale straw color, boy, that stuff certainly is stable, isn't it?
I feel fairly sure that there is a simple solution to my problem, waiting out there to be noticed, so please keep the suggestions coming.
cheers
rtch
On 10 Jan 2006 at 19:02, r.sims-at-auckland.ac.nz wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Here's a problem for the New Year: } } A couple of months ago, I repumped in situ my chronically leaky PGT Be-window EDS } detector, and ran 50 deg C air into the dewar while re-evacuating using the chamber } vacuum via a hose from the sample introduction port of the JEOL 840 on which the } detector is mounted. } } The re-evacuation worked well, in that the LN2 consumption is now back to normal, but } after recooling and switching on the bias again, there was no appreciable low-energy } response -- no visible Cu L peak at all! } } I realised that grease from the O-ring sliding seal through which the tube of the detector } enters the chamber had melted and run down onto the Be window. Cleaning of the } window with Freon restored the Cu L intensity, and all was well, until it became obvious } that the window was re-oiling again at a much faster rate than ever before. } } Because we do quantitative mineral analysis, I keep a good handle on the window } condition, evidenced by both Cu L and Na K responses. I have been using this detector } for about five years and not found it neccesary to clean the window before, but now the } Na response halves in a few weeks. Recleaning with Freon restores the response } again. } } In an effort to fix this, I have:- } } - changed the rotary pump oil } - installed an alumina-pellet foreline trap } - changed the DP oil (Santovac, but the old charge was still light-colored) } } but still the darned window is oiling up. } } I'm running out of ideas. It seems that either the chamber atmosphere has become } markedly oilier than before, or the detector window is now running colder than before. } } My money is on the latter, as it seems too much of a coincidence for some change in } the back-streaming performance of the 840 to have occurred at the same time as the } repumping of the detector, but I can't see what might have happened. } } Anyone got any suggestions? } } Happy New Year } } rtch } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand } } } ==============================Original Headers============================== } 16, 26 -- From r.sims-at-auckland.ac.nz Tue Jan 10 19:00:56 2006 } 16, 26 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) } 16, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0B10td7001103 } 16, 26 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 19:00:56 -0600 } 16, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) } 16, 26 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id F369034958 } 16, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 14:00:53 +1300 (NZDT) } 16, 26 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) } 16, 26 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) } 16, 26 -- with ESMTP id 31308-14 for {microscopy-at-microscopy.com} ; } 16, 26 -- Wed, 11 Jan 2006 14:00:46 +1300 (NZDT) } 16, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) } 16, 26 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 29EF235637 } 16, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 13:59:05 +1300 (NZDT) } 16, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } 16, 26 -- To: microscopy-at-microscopy.com } 16, 26 -- Date: Wed, 11 Jan 2006 13:59:04 +1300 } 16, 26 -- MIME-Version: 1.0 } 16, 26 -- Subject: Oil on detector window } 16, 26 -- Message-ID: {43C50F28.11970.13D9DFF-at-localhost} } 16, 26 -- Priority: normal } 16, 26 -- X-mailer: Pegasus Mail for Windows (4.21c) } 16, 26 -- Content-type: text/plain; charset=US-ASCII } 16, 26 -- Content-transfer-encoding: 7BIT } 16, 26 -- Content-description: Mail message body } 16, 26 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz } ==============================End of - Headers==============================
==============================Original Headers============================== 15, 27 -- From r.sims-at-auckland.ac.nz Wed Jan 11 17:09:46 2006 15, 27 -- Received: from groucho.itss.auckland.ac.nz (groucho.itss.auckland.ac.nz [130.216.190.11]) 15, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0BN9jBt024828 15, 27 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 17:09:46 -0600 15, 27 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) 15, 27 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 3333B352E7 15, 27 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 12:09:44 +1300 (NZDT) 15, 27 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) 15, 27 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 15, 27 -- with ESMTP id 13091-10 for {microscopy-at-microscopy.com} ; 15, 27 -- Thu, 12 Jan 2006 12:09:44 +1300 (NZDT) 15, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 15, 27 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 0E039343ED 15, 27 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 12:09:44 +1300 (NZDT) 15, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 15, 27 -- To: microscopy-at-microscopy.com 15, 27 -- Date: Thu, 12 Jan 2006 12:12:02 +1300 15, 27 -- MIME-Version: 1.0 15, 27 -- Subject: More about oil on detector window 15, 27 -- Message-ID: {43C64792.29976.BC74A1-at-localhost} 15, 27 -- Priority: normal 15, 27 -- In-reply-to: {200601110102.k0B12645002985-at-ns.microscopy.com} 15, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 15, 27 -- Content-type: text/plain; charset=US-ASCII 15, 27 -- Content-transfer-encoding: 7BIT 15, 27 -- Content-description: Mail message body 15, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
Dear Ritch, Does the snout of the EDS detector feel cool to the touch, when you vent the chamber? (Use the back of your hand to test it.) I had one detector that oiled very badly and it felt cool to the touch, so it acted as an oil trap for the chamber. I sent that detector to be completely rebuilt as a light element, high resolution detector and that problem has vanished, so it was a condition of the detector, not the SEM, since the SEM was not touched. It may be that the insulation between the liquid nitrogen or the cooled components and the snout of the detector has changed with your warm-up and re-pump. I am wondering if the copper braid that conducts the cool from the dewar to the nose of the detector has moved or is touching something. None of this helps you in any way. You can clean the Be-window detector with a trickle of pure ethanol or iso-propanol in the SEM, if you can get at it. ----- Original Message ----- X-from: {r.sims-at-auckland.ac.nz} To: {mager-at-interchange.ubc.ca} Sent: Wednesday, January 11, 2006 3:13 PM
Ritch, This is a long shot and you probably would have noticed the symptoms, but here goes:
I had one 840 that kept being "burped" first thing in the morning, but not after that. The key to finding the problem was that the LEDs on the vacuum control all cycled correctly, but it took me a while (since it only happened once a day) to notice that the actual valve operation for V2 (at the base of the second DP) was delayed just long enough to burp the DPs. Turns out the solenoid valve that operated V2 had Apiezon in it (a lot) and after sitting overnight would stick for about 2 seconds, only milliseconds longer than the delay for V5 to rough the load lock. This problem had been intermittent for years before I took on the service contract, but things work fine, now.
Perhaps you have a similar timing problem. Of course, the most obvious thing was that the DPs dumped and you don't seem to have had anything that severe happening.
Another thought: when you cleaned the DPs, did you also remove and clean the ballast tank located between them? Perhaps it has become too contaminated and is now backstreaming since there is only one DP between it and the chamber.
Do you have any additional gauging to monitor the actual pressure in the chamber? There might be some clues there, also. I'm thinking along the lines of a slightly leaky V4.
One last thought: Is the water flowing in the correct direction? An accidental reversal puts warm water running through the water baffles and virtually always results in oil on the EDS detector.
Best of luck, Ken Converse
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: r.sims-at-auckland.ac.nz [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, January 11, 2006 6:12 PM To: kenconverse-at-qualityimages.biz
Many thanks for the replies and suggestions so far, however, please note that this is a new condition.
For the past five years the oiling has not been a problem, and it has started to occur only since the repumping operation described below in tedious detail.
There have been no changes to the system apart from the three steps taken, one at a time, in an attempt to fix the problem (changing the RP oil; installing the foreline trap; and changing the DP oil), and they have made no difference.
I think it would be too much of a coincidence for the cause of the oiling to be unrelated to the repumping operation, so I think that complex cures such as installing LN2 traps, plasma cleaning, complete stripping and cleaning, etc, do not address the question, which is "what has changed to accelerate the detector window oiling so greatly, and how can I get back to the previous state?"
I had thought that perhaps a drop of melted grease might have dripped into the DP, and be decomposing in the Santovac, which propelled me to change the Santovac, but I now realise that this is very unlikely to be the case, as the DP is not directly beneath the sample chamber, and even if it were, such a drop of molten grease would not make it through the water-cooled baffle above the DP. Plus, of course, the fact that very thoroughly cleaning both of the DPs and both of their water-cooled baffles has made no difference at all to the problem.
Incidentally, the Santovac, after five years' continuous operation, was only a pale straw color, boy, that stuff certainly is stable, isn't it?
I feel fairly sure that there is a simple solution to my problem, waiting out there to be noticed, so please keep the suggestions coming.
cheers
rtch
On 10 Jan 2006 at 19:02, r.sims-at-auckland.ac.nz wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Here's a problem for the New Year: } } A couple of months ago, I repumped in situ my chronically leaky PGT Be-window EDS } detector, and ran 50 deg C air into the dewar while re-evacuating using the chamber } vacuum via a hose from the sample introduction port of the JEOL 840 on which the } detector is mounted. } } The re-evacuation worked well, in that the LN2 consumption is now back to normal, but } after recooling and switching on the bias again, there was no appreciable low-energy } response -- no visible Cu L peak at all! } } I realised that grease from the O-ring sliding seal through which the tube of the detector } enters the chamber had melted and run down onto the Be window. Cleaning of the } window with Freon restored the Cu L intensity, and all was well, until it became obvious } that the window was re-oiling again at a much faster rate than ever before. } } Because we do quantitative mineral analysis, I keep a good handle on the window } condition, evidenced by both Cu L and Na K responses. I have been using this detector } for about five years and not found it neccesary to clean the window before, but now the } Na response halves in a few weeks. Recleaning with Freon restores the response } again. } } In an effort to fix this, I have:- } } - changed the rotary pump oil } - installed an alumina-pellet foreline trap } - changed the DP oil (Santovac, but the old charge was still light-colored) } } but still the darned window is oiling up. } } I'm running out of ideas. It seems that either the chamber atmosphere has become } markedly oilier than before, or the detector window is now running colder than before. } } My money is on the latter, as it seems too much of a coincidence for some change in } the back-streaming performance of the 840 to have occurred at the same time as the } repumping of the detector, but I can't see what might have happened. } } Anyone got any suggestions? } } Happy New Year } } rtch } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand } } } ==============================Original Headers============================== } 16, 26 -- From r.sims-at-auckland.ac.nz Tue Jan 10 19:00:56 2006 } 16, 26 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) } 16, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0B10td7001103 } 16, 26 -- for {microscopy-at-microscopy.com} ; Tue, 10 Jan 2006 19:00:56 -0600 } 16, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) } 16, 26 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id F369034958 } 16, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 14:00:53 +1300 (NZDT) } 16, 26 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) } 16, 26 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) } 16, 26 -- with ESMTP id 31308-14 for {microscopy-at-microscopy.com} ; } 16, 26 -- Wed, 11 Jan 2006 14:00:46 +1300 (NZDT) } 16, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) } 16, 26 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 29EF235637 } 16, 26 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 13:59:05 +1300 (NZDT) } 16, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } 16, 26 -- To: microscopy-at-microscopy.com } 16, 26 -- Date: Wed, 11 Jan 2006 13:59:04 +1300 } 16, 26 -- MIME-Version: 1.0 } 16, 26 -- Subject: Oil on detector window } 16, 26 -- Message-ID: {43C50F28.11970.13D9DFF-at-localhost} } 16, 26 -- Priority: normal } 16, 26 -- X-mailer: Pegasus Mail for Windows (4.21c) } 16, 26 -- Content-type: text/plain; charset=US-ASCII } 16, 26 -- Content-transfer-encoding: 7BIT } 16, 26 -- Content-description: Mail message body } 16, 26 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz } ==============================End of - Headers==============================
==============================Original Headers============================== 15, 27 -- From r.sims-at-auckland.ac.nz Wed Jan 11 17:09:46 2006 15, 27 -- Received: from groucho.itss.auckland.ac.nz (groucho.itss.auckland.ac.nz [130.216.190.11]) 15, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0BN9jBt024828 15, 27 -- for {microscopy-at-microscopy.com} ; Wed, 11 Jan 2006 17:09:46 -0600 15, 27 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) 15, 27 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 3333B352E7 15, 27 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 12:09:44 +1300 (NZDT) 15, 27 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) 15, 27 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 15, 27 -- with ESMTP id 13091-10 for {microscopy-at-microscopy.com} ; 15, 27 -- Thu, 12 Jan 2006 12:09:44 +1300 (NZDT) 15, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 15, 27 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 0E039343ED 15, 27 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 12:09:44 +1300 (NZDT) 15, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 15, 27 -- To: microscopy-at-microscopy.com 15, 27 -- Date: Thu, 12 Jan 2006 12:12:02 +1300 15, 27 -- MIME-Version: 1.0 15, 27 -- Subject: More about oil on detector window 15, 27 -- Message-ID: {43C64792.29976.BC74A1-at-localhost} 15, 27 -- Priority: normal 15, 27 -- In-reply-to: {200601110102.k0B12645002985-at-ns.microscopy.com} 15, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 15, 27 -- Content-type: text/plain; charset=US-ASCII 15, 27 -- Content-transfer-encoding: 7BIT 15, 27 -- Content-description: Mail message body 15, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
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==============================Original Headers============================== 36, 23 -- From kenconverse-at-qualityimages.biz Thu Jan 12 09:30:50 2006 36, 23 -- Received: from qualityimages.biz (dpmail16.doteasy.com [65.61.209.16]) 36, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0CFUnmn022474 36, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Jan 2006 09:30:49 -0600 36, 23 -- Received: from Ken [68.64.242.101] by qualityimages.biz with ESMTP 36, 23 -- (SMTPD32-8.05) id A61691000C4; Thu, 12 Jan 2006 07:30:30 -0800 36, 23 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 36, 23 -- To: {r.sims-at-auckland.ac.nz} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 36, 23 -- Subject: RE: [Microscopy] More about oil on detector window 36, 23 -- Date: Thu, 12 Jan 2006 10:30:26 -0500 36, 23 -- Message-ID: {003701c6178d$2273cd80$6501a8c0-at-Ken} 36, 23 -- MIME-Version: 1.0 36, 23 -- Content-Type: text/plain; 36, 23 -- charset="us-ascii" 36, 23 -- X-Priority: 3 (Normal) 36, 23 -- X-MSMail-Priority: Normal 36, 23 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 36, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 36, 23 -- In-Reply-To: {200601112312.k0BNCQVD030081-at-ns.microscopy.com} 36, 23 -- Importance: Normal 36, 23 -- X-IMSTrailer: __IMail_7__ 36, 23 -- Content-Transfer-Encoding: 8bit 36, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0CFUnmn022474 ==============================End of - Headers==============================
Transmission Electron Microscopy (http://www.collegeofmicroscopy.com/courses/17.asp)
Both are hands-on, introductory level courses taught using the latest equipment available. Please follow the links provided for further details and registration information.
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
==============================Original Headers============================== 11, 27 -- From eschumacher-at-mccrone.com Thu Jan 12 14:13:16 2006 11, 27 -- Received: from pgp.mccrone.com (McCrone-Associates-1051626.cust-rtr.ameritech.net [68.23.63.122]) 11, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0CKDGdt003847 11, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Jan 2006 14:13:16 -0600 11, 27 -- Received: from pgp.mccrone.com (localhost [127.0.0.1]) 11, 27 -- by pgp.mccrone.com (Postfix) with ESMTP id 4FC601A800B 11, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Jan 2006 14:13:17 -0600 (CST) 11, 27 -- Received: from MCCRONEMSG.tmg.mccrone.com ([192.168.101.20]) 11, 27 -- by pgp.mccrone.com (PGP Universal service); 11, 27 -- Thu, 12 Jan 2006 14:13:17 -0600 11, 27 -- X-PGP-Universal: processed 11, 27 -- Content-class: urn:content-classes:message 11, 27 -- MIME-Version: 1.0 11, 27 -- Content-Type: text/plain; 11, 27 -- charset="US-ASCII" 11, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.6944.0 11, 27 -- Subject: Short Course Announcement: Scientific Imaging, Transmission Electron Microscopy 11, 27 -- Date: Thu, 12 Jan 2006 14:13:06 -0600 11, 27 -- Message-ID: {2A27CE0EC2A5C748974EA513DFB285A74C30F7-at-MCCRONEMSG.tmg.mccrone.com} 11, 27 -- X-MS-Has-Attach: 11, 27 -- X-MS-TNEF-Correlator: 11, 27 -- Thread-Topic: Short Course Announcement: Scientific Imaging, Transmission Electron Microscopy 11, 27 -- Thread-Index: AcYXtJk/0FDZxb4QTf2pOACZsYbNJQ== 11, 27 -- From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} 11, 27 -- To: {Microscopy-at-MSA.Microscopy.Com} 11, 27 -- Content-Transfer-Encoding: 8bit 11, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0CKDGdt003847 ==============================End of - Headers==============================
Does anyone know of a reasonable alternative to the venerable Zero Stat static taming guns? We use them often but there's just got to be a better way. I used to smuggle squirt guns onto the school bus that were made better (the good old days), and they didn't cost over $100 for about $1 worth of components.
Thanks and Happy New Year to all.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 9, 23 -- From TindallR-at-missouri.edu Thu Jan 12 16:59:22 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 (8.12.11/8.12.8) with ESMTP id k0CMxMkH015190 9, 23 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 16:59:22 -0600 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 -- Thu, 12 Jan 2006 16:59:22 -0600 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: Static guns? 9, 23 -- Date: Thu, 12 Jan 2006 16:59:21 -0600 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849CB7-at-UM-EMAIL09.um.umsystem.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Static guns? 9, 23 -- Thread-Index: AcYXy9ND13Q0Mk2eRQC7DwCisMtGXA== 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 12 Jan 2006 22:59:22.0766 (UTC) FILETIME=[D3E862E0:01C617CB] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0CMxMkH015190 ==============================End of - Headers==============================
Job Opening #B005608; Lab Technician/Engineer at IBM
There is an opening at the Lab Technician/Engineer level within the Materials Characterization and Analysis Group in the Research Division of IBM at the Almaden Research Center in San Jose, California. The job is a long term (three years) supplemental position with benefits. A B.S. in material science or equivalent fields, or successful completion of a two year college program plus experience, is required.
This position involves technical support in an advanced electron microscopy laboratory which emphasizes transmission electron microscopy. The technician works as a team member with professional (Ph.D.) microscopists and other scientists in a research laboratory. Duties involve primarily sample preparation methods including the conventional grinding, polishing, and ion milling techniques, focused ion beam (FIB ) method, as well as Microtome, etc. In addition, the candidate must be able to operate and perform routine maintenance on specimen preparation equipment, interact with customers (professional scientists), and prepare reports. The candidate must be able to work independently within set priorities, to keep abreast of new advances, and to interact smoothly within a team.
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Dr. Philip M. Rice IBM Research Division Almaden Research Center 650 Harry Road, K19B/D1 tel: (408) 927-1442 fax: (408) 927-2100 email: pmrice-at-almaden.ibm.com
Dr. Philip M. Rice IBM Research Division Almaden Research Center 650 Harry Road, K19B/D1 San Jose, CA 95120-6099 tel: (408) 927-1442 fax: (408) 927-2100 e-mail: pmrice-at-almaden.ibm.com
==============================Original Headers============================== 10, 26 -- From pmrice-at-almaden.ibm.com Thu Jan 12 17:19:37 2006 10, 26 -- Received: from e6.ny.us.ibm.com (e6.ny.us.ibm.com [32.97.182.146]) 10, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0CNJaZ0024520 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 17:19:36 -0600 10, 26 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 10, 26 -- by e6.ny.us.ibm.com (8.12.11/8.12.11) with ESMTP id k0CNJZhi023925 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 18:19:35 -0500 10, 26 -- Received: from d01av03.pok.ibm.com (d01av03.pok.ibm.com [9.56.224.217]) 10, 26 -- by d01relay04.pok.ibm.com (8.12.10/NCO/VERS6.8) with ESMTP id k0CNJZBB128016 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 18:19:35 -0500 10, 26 -- Received: from d01av03.pok.ibm.com (loopback [127.0.0.1]) 10, 26 -- by d01av03.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k0CNJZ9b030875 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 18:19:35 -0500 10, 26 -- Received: from d01ml605.pok.ibm.com (d01ml605.pok.ibm.com [9.56.227.91]) 10, 26 -- by d01av03.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k0CNJZ8m030872 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 12 Jan 2006 18:19:35 -0500 10, 26 -- Subject: TEM - Job opening for a TEM Specimen Preparation Lab Technician 10, 26 -- To: microscopy-at-microscopy.com 10, 26 -- X-Mailer: Lotus Notes Release 6.0.2CF1 June 9, 2003 10, 26 -- Message-ID: {OF50DE3E5B.2E1B3545-ON882570F4.007FA844-882570F4.008020F8-at-us.ibm.com} 10, 26 -- From: Philip Rice {pmrice-at-almaden.ibm.com} 10, 26 -- Date: Thu, 12 Jan 2006 15:19:30 -0800 10, 26 -- X-MIMETrack: Serialize by Router on D01ML605/01/M/IBM(Release 7.0HF90 | November 16, 2005) at 10, 26 -- 01/12/2006 18:19:34 10, 26 -- MIME-Version: 1.0 10, 26 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I am trying to permeabilize some Haemophilus influenzae fixed in 2% paraformaldehyde + 0.2% glutaraldehyde so I can immunostain an intracellular protein. My first attempts to permeabilize with Triton X-100 were surprising unsuccessful. I tried both literature searches and googling this topic but a search of "bacteria" + "permeabilization" pulls up thousands on non-germane citations. I am interested in comments from anybody with expertise in permeabilization, especially in regard to the bacterial target. Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
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Email: aetmicro-at-optonline.net Name: Andrew Thelian
Title-Subject: [Filtered] Philips EM 300 beam issue
Question: Hi All,
I have hit yet another bump in the road...
I was having a brightness issue last week that i thought might be resolved with cleaning the wehnelt assembly...
Now when i energize the filament i get a very small circle on the view screen...it doesnt appear green it looks like a dim reddish color...it also seems to respond to turning the filament knob...
the condenser and deflection knobs dont cause any change...
I have removed all apertures... to see if there was a beam any where ... but no luck...
This Question was submitted to Ask-A-Microscopist by (dustin.adams-at-xerox.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, January 12, 2006 at 17:11:24 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both dustin.adams-at-xerox.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Hello, I am going to be training soon to become a SEM microscopist for my company, and was curious to see what the salary amounts are for a typical person in my position. I have tried to look for this info online, but wasn't really able to find anything. If someone could provide that information or point me in the right direction; I would greatly appreciate it.
What you see is the light from filament, not e-beam. Whenever electron gun was disturbed, in your case for Wehnelt cleaning, gun tilt must be re-aligned, and perhaps gun shift too. Both these alignments are mechanical on Phil. EM300. Users manual will help if TEM is functioning properly.
Green light or not, you must be able to tell whether HT is present by watching emission current, and emission meter behavior: a) when HT turned on/off; b) emission setting changed while HT is on; c) and HV setting changed while HT is on.
If HT is present, if filament glows, and if emission current behaves normally, do the following: a) select lowest magnification - I believe it is SC (scan) position; b) move all apertures from the beam pass; c) if no beam yet, turn lens currents off one by one and you must see the beam. Be careful not to burn the screen (keep filament under-saturated). Just turning off C1 and C2 will likely bring the beam back, and then work from that point on.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: {aetmicro-at-optonline.net} To: {vitalylazar-at-att.net} Sent: Thursday, January 12, 2006 9:23 PM
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Email: secr-at-eurmicsoc.org Name: Nick Schryvers
Organization: EMS
Title-Subject: [Filtered] EMS scholarships for IMC16 Msg 2006004
Question: The European Microscopy Society is pleased to be able to offer 4 scholarships of 500 Euro each to young researchers in support of attending the 16th International Microscopy Congress (http://www.imc16.jp/) in Sapporo, Japan, from Sep. 3 till Sep. 8, 2006. Applications should be sent to the EMS secretary (secr-at-eurmicsoc.org) and include a CV and list of publications and copies of the submitted abstract(s) at IMC16. Proof of acceptance of the abstract(s) should follow as soon as these are available.
Deadline of the applications is February 1, 2006 (same date as abstract submission for IMC16)
Only EMS members are eligible and priority will be given to people under the age of 35.
This support was made possible through a generous offer from JEOL Europe (http://www.jeoleuro.com/).
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both secr-at-eurmicsoc.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: secr-at-eurmicsoc.org Name: Nick Schryvers
Organization: EMS
Title-Subject: [Filtered] Royal Microscopical Society ( RMS) Spring School in Electron Microscopy
Royal Microscopical Society ( RMS) Spring School in Electron Microscopy 27-31 March 2006 University of Leeds, UK. Course Organiser: Prof. Rik Brydson
The RMS School consists of a number of parallel courses covering most aspects of electron microscopy in both the physical and life sciences. Students can attend either 4 or 5 days - the 5 day course includes a day of principles of EM lectures for beginners or as a refresher. During the week, students will have the opportunity to choose to attend lectures and workshops from either course, to tailor it to their specific needs.
'All those involved in the organisation of this event are to be congratulated. The complexity of running a school of electron microscopy covering both life and materials sciences has been surpassed with remarkable success with participants, speakers and organisers equally satisfied with the end results.'
Pedro Costa - Participant Spring School in EM 2005
For more information or to book for this course please visit the RMS website: {http://www.rms.org.uk/event_em_school.shtml} http://www.rms.org.uk/event_em_school.shtml
Victoria Lee Conference Manager Royal Microscopical Society 37/38 St Clements, Oxford OX4 1AJ
I always used a sealed Polonium-210 beta source for static problems - Nuclepore Corp used to make a Polonium-210 (sealed in beads) metal strip that had an open ladder arrangement at the top that you simply placed the Nuclepore filters on. It really reduced static. The only downside was the half-life of a few months that meant after a year or so you had to buy another. We also had Zero-Stat gun, but that wasn't used as it simply didn't work. The Nuclepore device seems to have been dropped now, but you can buy a brush that has Polonium-210 trapped within the bristles that should work on similar lines (NRD - StaticMaster Anti-Static Brush - 1" or 3") :
I know that photographers use them to reduce static on film. I've never tried this specific brush though as I have now moved out of this field. Our histologist used something similar on microtome surfaces when sectioning I remember.
Also have a look at ionisers like : http://www.etherton.co.uk/index.html http://www.2spi.com/catalog/photo/antistat.html
Alternatively you can spray a conducting coating onto the plastic to eliminate static (e.g. the Duron anti-static spray sold by Agar Scientific that "even allows uncoated samples to be viewed under 'low resolution' in an SEM").
There's also the anti-static wrist-strap (or just touching the metal earth of a plugged in PC case or wall socket screw head). Plus breathing on the plastic really removes the static (or earthing the material if it conducts), but that suits plastic record player Perspex covers more than scientific specimens. Keeping the room %RH constant and not going too low will also help. Some labs have a full antistatic grounding system with furniture, mats etc.. all 'grounded' with controlled %RH and temperatures, but we got by with antistatic mats, higher humidity and the polonium-210. We avoided using gloves as much as possible, using fine metal forceps for handling filters (I don't think we bothered with 'antistatic' forceps). Antistatic gloves weren't much use as we needed disposable ones. We used very thin sensitive clear plastic gloves that were probably something like Polyethylene, although they probably did have some static problems. Our Nuclepore filter samples were generally glassfibres recovered from lungs (fibre durability + toxicology studies) or aerosol particles, and obviously when using aqueous samples static was only a problem with the clean filter prior to filtration or after drying.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {TindallR-at-missouri.edu} To: {keith.morris-at-ucl.ac.uk} Sent: Thursday, January 12, 2006 11:04 PM
I'll send you a PDF of the salary survey article that appeared in Microscopy Today last year. Also, if you are going to be a microscopist (congratulations!) you should sign up for a free subscription to Microscopy Today at http://www.microscopy-today.com.
Ron Anderson, Editor
dustin.adams-at-xerox.com wrote:
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==============================Original Headers============================== 6, 20 -- From microscopytoday-at-tampabay.rr.com Fri Jan 13 08:32:19 2006 6, 20 -- Received: from ms-smtp-03.tampabay.rr.com (ms-smtp-03-smtplb.tampabay.rr.com [65.32.5.133]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DEWIuY013660 6, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Jan 2006 08:32:18 -0600 6, 20 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 6, 20 -- by ms-smtp-03.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k0DEWDul022696; 6, 20 -- Fri, 13 Jan 2006 09:32:17 -0500 (EST) 6, 20 -- Message-ID: {43C7B9EC.8030201-at-tampabay.rr.com} 6, 20 -- Date: Fri, 13 Jan 2006 09:32:12 -0500 6, 20 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 6, 20 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 6, 20 -- X-Accept-Language: en-us, en 6, 20 -- MIME-Version: 1.0 6, 20 -- To: dustin.adams-at-xerox.com, Listserver {Microscopy-at-MSA.Microscopy.Com} 6, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: SEM microscopist salaries 6, 20 -- References: {200601130224.k0D2Oc6V017874-at-ns.microscopy.com} 6, 20 -- In-Reply-To: {200601130224.k0D2Oc6V017874-at-ns.microscopy.com} 6, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 20 -- Content-Transfer-Encoding: 7bit 6, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Thanks for all the suggestions on static control! I learned about some new things to try for general use. I had forgotten about the polonium strip StaticMaster brushes from my days locked in a photo darkroom, but this may be a good alternative to these outrageously overpriced Zero Stat guns that don't work nearly as well as they did a few years ago.
Those StaticMaster brushes used to strike fear into us when we read the disposal instructions, which were something like "seal this strip inside a deep, deep mine and don't go anywhere near it for 5000 years", although we could use them in the darkroom with no special protection. They did work, though, except you couldn't spark your co-workers with them and make them squeal.
By the way, our most common use for these guns is somewhat trivial: getting grids down off the lid of the petri dish, where static electricity glues them, especially in winter. (Tamara, I agree with you about the static in New Mexico---I still have scars from getting zapped every time I'd get out of my car in Las Cruces!). We use the Diatome de-ionizer for microtomy, which works well.
Thanks again!
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 11, 23 -- From TindallR-at-missouri.edu Fri Jan 13 08:50:02 2006 11, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 11, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DEo1kl023115 11, 23 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 08:50:01 -0600 11, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 11, 23 -- Fri, 13 Jan 2006 08:49:59 -0600 11, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 11, 23 -- Content-class: urn:content-classes:message 11, 23 -- MIME-Version: 1.0 11, 23 -- Content-Type: text/plain; 11, 23 -- charset="us-ascii" 11, 23 -- Subject: Static! 11, 23 -- Date: Fri, 13 Jan 2006 08:49:58 -0600 11, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849CB9-at-UM-EMAIL09.um.umsystem.edu} 11, 23 -- X-MS-Has-Attach: 11, 23 -- X-MS-TNEF-Correlator: 11, 23 -- Thread-Topic: Static! 11, 23 -- Thread-Index: AcYYUKGubaVRxOz9TI+TEu+84a1Tdg== 11, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 11, 23 -- To: {microscopy-at-microscopy.com} 11, 23 -- X-OriginalArrivalTime: 13 Jan 2006 14:49:59.0341 (UTC) FILETIME=[A05B0DD0:01C61850] 11, 23 -- Content-Transfer-Encoding: 8bit 11, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0DEo1kl023115 ==============================End of - Headers==============================
You want static??!!! Try Manitoba in February. When it's -30C there's no such thing as moisture in the air. But then, not even Phil was willing to try Manitoba in June/July, even for real beer ;-) .
Great time to make formvar grids, though.
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 } Phone:204-789-3313 } Pager:204-931-9354 } Cell:204-781-1502 } Fax:204-789-3926
==============================Original Headers============================== 12, 20 -- From paul_hazelton-at-umanitoba.ca Fri Jan 13 09:59:12 2006 12, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 12, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DFwxMP001111 12, 20 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 09:59:01 -0600 12, 20 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 12, 20 -- (authenticated bits=0) 12, 20 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k0DFwtYb015625; 12, 20 -- Fri, 13 Jan 2006 09:58:56 -0600 (CST) 12, 20 -- Message-ID: {43C7CE3B.4080709-at-umanitoba.ca} 12, 20 -- Date: Fri, 13 Jan 2006 09:58:51 -0600 12, 20 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 12, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 12, 20 -- X-Accept-Language: en-us, en 12, 20 -- MIME-Version: 1.0 12, 20 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 12, 20 -- Subject: Re: [Microscopy] Static! 12, 20 -- References: {200601131453.k0DErNj8026940-at-ns.microscopy.com} 12, 20 -- In-Reply-To: {200601131453.k0DErNj8026940-at-ns.microscopy.com} 12, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed 12, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Definition of milli-second is the amount of time between hitting send and realizing there is an error, da? Of course I meant phospholipid, not lipoprotein when mentioning the outer and cytoplasmic membranes. Mark it up to trying to do e-mail when the brain is turned off. Should Ron Anderson decide that this is a train worth following for Microscopy Today and want to use my response to you, perhaps he could make the correction, please??!!
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 } Phone:204-789-3313 } Pager:204-931-9354 } Cell:204-781-1502 } Fax:204-789-3926
==============================Original Headers============================== 10, 20 -- From paul_hazelton-at-umanitoba.ca Fri Jan 13 09:59:41 2006 10, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DFxdMb001278 10, 20 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 09:59:40 -0600 10, 20 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 10, 20 -- (authenticated bits=0) 10, 20 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k0DFxcsx015957 10, 20 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 09:59:38 -0600 (CST) 10, 20 -- Message-ID: {43C7CE65.7000006-at-umanitoba.ca} 10, 20 -- Date: Fri, 13 Jan 2006 09:59:33 -0600 10, 20 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 10, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 10, 20 -- X-Accept-Language: en-us, en 10, 20 -- MIME-Version: 1.0 10, 20 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 10, 20 -- Subject: Re: [Microscopy] bacterial permeabilization - further 10, 20 -- References: {200601130008.k0D08WwZ005845-at-ns.microscopy.com} 10, 20 -- In-Reply-To: {200601130008.k0D08WwZ005845-at-ns.microscopy.com} 10, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed 10, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Very interesting question. Made more interesting to me by the fact that I was discussing this same topic this afternoon, only more generically, and certainly not in reference to H. influenza. The problem you are faced with is bacterial structure. H. influenza is a Gram negative microorganism. Gram negative cells have two lipoprotein layers - the outer and cytoplasmic membranes. In addition, you have a layer of peptidoglycan between the outer and cytoplasmic membranes. To make it even more fun, there is frequently a lypopolysaccharide layer around the whole thing. You have to penetrate the LPS, permeabilize the outer membrane, traverse the periplasmic space (the volume between the outer and cytoplasmic membranes), cross the peptidoglycan layer and then penetrate the inner membrane, all with gentle treatment to allow the membranes to reorganize after removal of the detergent you use for permeabilization. What it adds up to is that permeabilization will be difficult at best, and most likely impossible. Probably the only way to get something reliably into the cytoplasm would be by active transport.
Having said that, knowing what I do about the structure of prokaryotic cells, I confess to having tried to permeabilize and do pre-embedding labeling on E. coli expressing a protein of interest (another gram negative bacterium). Didn't expect it, but tried anyway. Hey, dogma schmogma, give it a try, right? That's how the serendipitous findings occur. Anyway, used triton X, used saponin. Ugly. Did not look good. No luck either way. So logic and dogma were right this time.
At the same time I did LR white embedding on cells from the same broth culture. Included osmium fixation. Went back with metaperiodate followed by hydrogen peroxide etching and then did indirect IEM with 12nm gold for the label. Beautiful preservation. Got highly significant labeling of the expressed protein in the E. coli and in the wild type bacterium from which the protein of interest had been cloned. In fact, the protein of interest was hypothesized to be membrane inserted on the basis of amino acid sequence and the labeling of the wild type cells was primarily associated with the cytoplasmic membrane. In short, I did minor modification to the standard LR white embedding protocol and it worked great. I would give that a try.
I would like to give a reference for the work, but we are still waiting for the student to finish his thesis, not to mention the paper. And he has already left for his post-doc. If you have any questions, call. I would be glad to discuss the procedure I used further.
Paul
==============================Original Headers============================== 9, 21 -- From paul_hazelton-at-umanitoba.ca Fri Jan 13 10:12:07 2006 9, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 9, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DGC78e019513 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 10:12:07 -0600 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 k0DGC684025968; 9, 21 -- Fri, 13 Jan 2006 10:12:06 -0600 (CST) 9, 21 -- Message-ID: {43C7D151.2010503-at-umanitoba.ca} 9, 21 -- Date: Fri, 13 Jan 2006 10:12:01 -0600 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: Microscopy Listserver {microscopy-at-microscopy.com} 9, 21 -- Subject: Re: [Microscopy] bacterial permeabilization 9, 21 -- References: {200601130008.k0D08WwZ005845-at-ns.microscopy.com} 9, 21 -- In-Reply-To: {200601130008.k0D08WwZ005845-at-ns.microscopy.com} 9, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 21 -- Content-Transfer-Encoding: 8bit 9, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0DGC78e019513 ==============================End of - Headers==============================
Hey! I would've been happy to go to Manitoba, even for real beer. Just kind of hard when I was busy fending off predatory administraitors. Gave up on that and just moved.
Phil
Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576
Hi, Dustin
We did a salary survey for the society in conjunction with Microscopy Today late last year. Contact Ron Anderson for details. (see email above).
Hope this is helpful, 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 08:25 PM 1/12/2006, dustin.adams-at-xerox.com wrote:
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==============================Original Headers============================== 13, 18 -- From bfoster-at-mme1.com Fri Jan 13 10:57:55 2006 13, 18 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 13, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DGvtFJ005902 13, 18 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 10:57:55 -0600 13, 18 -- Received: (qmail 27399 invoked from network); 13 Jan 2006 11:59:33 -0500 13, 18 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 13, 18 -- by enterprise.5starpro.com with SMTP; 13 Jan 2006 11:59:33 -0500 13, 18 -- Message-Id: {6.1.2.0.0.20060113105646.0273a880-at-mail.mme1.com} 13, 18 -- X-Sender: bfoster-at-mail.mme1.com 13, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.2.0 13, 18 -- Date: Fri, 13 Jan 2006 10:57:56 -0600 13, 18 -- To: dustin.adams-at-xerox.com, microscopy-at-microscopy.com 13, 18 -- From: Barbara Foster {bfoster-at-mme1.com} 13, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: SEM microscopist salaries 13, 18 -- In-Reply-To: {200601130225.k0D2PxKo021933-at-ns.microscopy.com} 13, 18 -- References: {200601130225.k0D2PxKo021933-at-ns.microscopy.com} 13, 18 -- Mime-Version: 1.0 13, 18 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
At 08:25 PM 1/12/2006, dustin.adams-at-xerox.com wrote:
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==============================Original Headers============================== 6, 18 -- From bfoster-at-mme1.com Fri Jan 13 11:03:21 2006 6, 18 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DH3KI6015095 6, 18 -- for {microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 11:03:20 -0600 6, 18 -- Received: (qmail 27830 invoked from network); 13 Jan 2006 12:04:58 -0500 6, 18 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 6, 18 -- by enterprise.5starpro.com with SMTP; 13 Jan 2006 12:04:58 -0500 6, 18 -- Message-Id: {6.1.2.0.0.20060113105606.026f6358-at-mail.mme1.com} 6, 18 -- X-Sender: bfoster-at-mail.mme1.com 6, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.2.0 6, 18 -- Date: Fri, 13 Jan 2006 11:03:21 -0600 6, 18 -- To: dustin.adams-at-xerox.com, microscopy-at-microscopy.com 6, 18 -- From: Barbara Foster {bfoster-at-mme1.com} 6, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: SEM microscopist salaries 6, 18 -- In-Reply-To: {200601130225.k0D2PxKo021933-at-ns.microscopy.com} 6, 18 -- References: {200601130225.k0D2PxKo021933-at-ns.microscopy.com} 6, 18 -- Mime-Version: 1.0 6, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I am looking at getting a deeply cooled (-30 C) high sensitivity, high resolution digital camera. I am considering the following cameras : Hamamatsu Orca-AG Retiga SRV Photometrics CoolSnap HQ or K4
Any comments (public or private) on your experiences would be appreciated.
One source has told me that they see little practical difference in the Orca (cooled to -30) vs the Retiga EXi (cooled to 25 below ambient). Comments about this statement are also welcome.
Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
McCrone Associates, Inc., an innovative modern analytical and research laboratory, seeks a highly motivated electron microscopy technician with training in ultramicrotomy for materials science. The successful candidate will support our Electron Optics Group in sample preparation for SEM and TEM, as well as instrument maintenance. Ability to carry out routine SEM and TEM imaging and x-ray analysis is also desired. For more information on The McCrone Group, please visit www.mccrone.com.
The ideal candidate will have a B.S. degree with coursework in electron microscopy and microtomy, or comparable experience. Compensation is commensurate with education, experience and responsibilities.
Applicant selected must be a U.S. Citizen, will be subject to a government security investigation, and must meet eligibility requirements for access to classified information. McCrone is an Equal Opportunity Employer.
If you have microtomy experience and are looking for an interesting and unique career opportunity in materials analysis, send your resume with a letter describing your experience and interests to:
Careers McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, Illinois 60559
FAX: 630-887-7417 - Attn: Human Resources E-mail: careers-at-mccrone.com (e-mail is preferred)
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
==============================Original Headers============================== 11, 27 -- From eschumacher-at-mccrone.com Fri Jan 13 12:14:59 2006 11, 27 -- Received: from pgp.mccrone.com (McCrone-Associates-1051626.cust-rtr.ameritech.net [68.23.63.122]) 11, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DIEwln001747 11, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Jan 2006 12:14:59 -0600 11, 27 -- Received: from pgp.mccrone.com (localhost [127.0.0.1]) 11, 27 -- by pgp.mccrone.com (Postfix) with ESMTP id DA9E61A800B 11, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Jan 2006 12:14:58 -0600 (CST) 11, 27 -- Received: from MCCRONEMSG.tmg.mccrone.com ([192.168.101.20]) 11, 27 -- by pgp.mccrone.com (PGP Universal service); 11, 27 -- Fri, 13 Jan 2006 12:14:58 -0600 11, 27 -- X-PGP-Universal: processed 11, 27 -- Content-class: urn:content-classes:message 11, 27 -- MIME-Version: 1.0 11, 27 -- Content-Type: text/plain; 11, 27 -- charset="US-ASCII" 11, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.6944.0 11, 27 -- Subject: Open Position: Electron Microscopy Technician 11, 27 -- Date: Fri, 13 Jan 2006 12:14:48 -0600 11, 27 -- Message-ID: {2A27CE0EC2A5C748974EA513DFB285A74C3103-at-MCCRONEMSG.tmg.mccrone.com} 11, 27 -- X-MS-Has-Attach: 11, 27 -- X-MS-TNEF-Correlator: 11, 27 -- Thread-Topic: Open Position: Electron Microscopy Technician 11, 27 -- Thread-Index: AcYYbT0QYMn4lwyUQBa7kWOedjOuMA== 11, 27 -- From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} 11, 27 -- To: {Microscopy-at-MSA.Microscopy.Com} 11, 27 -- Content-Transfer-Encoding: 8bit 11, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0DIEwln001747 ==============================End of - Headers==============================
What you are seeing is the projected image of the hot filament (light). This means that although you have a hole down the column the electron gun is out of alignment. May I suggest the following.
At no more than 60kV switch off all the lenses and look for a very bright electron beam by adjusting the gun alignment controls. Once you find the beam switch each lens on one at a time, stepping back if the beam disappears.
Good luck
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: {aetmicro-at-optonline.net} To: {protrain-at-emcourses.com} Sent: Friday, January 13, 2006 2:22 AM
Group,
Our University in an effort of conservation of energy is making changes as to the HVAC conditions they can provide us. Requests as to equipment shuts downs during non use hours, shuting off computers every time not in use, and increasing lab temperatures beyound 78F are being asked for. Looking for documented case studies pro or con and anyones experience they would be willing to share offline on this issue. Would also like to hear from vendors as to desired environments for SEM and related equipment as well as stored chemicals.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 7, 20 -- From rbeavers-at-mail.smu.edu Fri Jan 13 15:12:52 2006 7, 20 -- Received: from s31xe5.systems.smu.edu (s31xe5.systems.smu.edu [129.119.70.74]) 7, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0DLCpEo022529 7, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Jan 2006 15:12:51 -0600 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 7, 20 -- Content-class: urn:content-classes:message 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="iso-8859-1" 7, 20 -- Subject: Lab Facilities Temperature 7, 20 -- Date: Fri, 13 Jan 2006 15:12:51 -0600 7, 20 -- Message-ID: {3A9F6E299461A44483961A57175598A703402D0F-at-s31xe5.systems.smu.edu} 7, 20 -- X-MS-Has-Attach: 7, 20 -- X-MS-TNEF-Correlator: 7, 20 -- Thread-Topic: Lab Facilities Temperature 7, 20 -- Thread-Index: AcYYhhysHdUwLzATSNSFiQlctfoZlA== 7, 20 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 7, 20 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 7, 20 -- Content-Transfer-Encoding: 8bit 7, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0DLCpEo022529 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mswift-at-bunham.org) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, January 13, 2006 at 18:18:43 ---------------------------------------------------------------------------
Email: mswift-at-bunham.org Name: Mark Swift
Organization: Burnham Institute
Education: Graduate College
Location: San Diego, CA
Question: We have a Tecnai 12 from FEI, and we would like to know: How do we ensure that in Low Dose our Focus Substates 1 and 2 are on the tilt axis when Alpha angle is not 0 degrees?
I am seeking advice on obtaining a low cost polisher for SEM sample preparation. The samples will be coated thin metal cross-sections embedded in epoxy. The cross-sections will be analyzed by SEM to determine coating thickness. I have a diamond wet saw but I need a polisher but have very little funds available. I found a used Buehler Minimet polisher in my price range ~$1K. I am not familiar with this unit. I have used bigger units like the Buehler Powerpro and the Leco SS-2000. Is the difference mainly a matter of speed and sample size? I plan on doing only about one sample a week. Any advice will be appreciated. Thanks,
Scott LeMaster PPG Industries Inc. lemaster-at-ppg.com
==============================Original Headers============================== 4, 23 -- From lemaster-at-ppg.com Mon Jan 16 09:31:25 2006 4, 23 -- Received: from SGOFSMTP02.nac.ppg.com (smtpout.ppg.com [141.189.251.4]) 4, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0GFVMxY018188 4, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Jan 2006 09:31:24 -0600 4, 23 -- Received: from SGOFUSR24.nac.ppg.com ([10.49.120.133]) by SGOFSMTP02.nac.ppg.com with Microsoft SMTPSVC(5.0.2195.6713); 4, 23 -- Mon, 16 Jan 2006 10:31:20 -0500 4, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 4, 23 -- Content-Class: urn:content-classes:message 4, 23 -- MIME-Version: 1.0 4, 23 -- Content-Type: text/plain; 4, 23 -- charset="us-ascii" 4, 23 -- Subject: SEM advice on low cost polisher 4, 23 -- Date: Mon, 16 Jan 2006 10:31:20 -0500 4, 23 -- Message-ID: {B92A132E126C4E45B3AAC00B5623D3E7015AF8CA-at-sgofusr24.nac.ppg.com} 4, 23 -- X-MS-Has-Attach: 4, 23 -- X-MS-TNEF-Correlator: 4, 23 -- Thread-Topic: SEM advice on low cost polisher 4, 23 -- thread-index: AcYaseX2g8/b/IiKQ069aQDRzXKGcA== 4, 23 -- From: "LeMaster, J. Scott" {lemaster-at-ppg.com} 4, 23 -- To: {Microscopy-at-microscopy.com} 4, 23 -- X-OriginalArrivalTime: 16 Jan 2006 15:31:20.0666 (UTC) FILETIME=[E69473A0:01C61AB1] 4, 23 -- Content-Transfer-Encoding: 8bit 4, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0GFVMxY018188 ==============================End of - Headers==============================
Scott: on the MiniMet, the sample moves and the polishing media stays still. This tool can take up to a 2 inch potted sample and uses 4-inch PSA grinding/polishing media on glass discs. It was designed to do rough and fine polish and it does that pretty well. On this tool, you drill a small blind hole in the top of your sample. This is where a spring-loaded arm fits to move the sample in an random orbital motion. You have some control over the pressure of the sample into the media and the speed of rotation. It can run unattended. This tool might be just what you need for one sample a week, if you can cut almost to the area of interest and then polish the rest of the way. On used tools, the spring force is usually the first thing to go due to rust, so be aware.
lemaster-at-ppg.com wrote:
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-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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============================== 5, 23 -- From r-holdford-at-ti.com Mon Jan 16 10:43:49 2006 5, 23 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0GGhmBh028324 5, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Jan 2006 10:43:48 -0600 5, 23 -- Received: from dlep30.itg.ti.com ([157.170.170.32]) 5, 23 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k0GGh3mi021873; 5, 23 -- Mon, 16 Jan 2006 10:43:32 -0600 (CST) 5, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 5, 23 -- by dlep30.itg.ti.com (8.12.11/8.12.11) with ESMTP id k0GGf6pY012416; 5, 23 -- Mon, 16 Jan 2006 10:41:07 -0600 (CST) 5, 23 -- Message-ID: {43CBCCA2.2060902-at-ti.com} 5, 23 -- Date: Mon, 16 Jan 2006 10:41:06 -0600 5, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 5, 23 -- Organization: SC Packaging Development -- FA Development 5, 23 -- User-Agent: Mozilla Thunderbird 0.8 (Windows/20040913) 5, 23 -- X-Accept-Language: en-us, en 5, 23 -- MIME-Version: 1.0 5, 23 -- To: lemaster-at-ppg.com, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 23 -- Subject: Re: [Microscopy] SEM advice on low cost polisher 5, 23 -- References: {200601161533.k0GFXXfg018559-at-ns.microscopy.com} 5, 23 -- In-Reply-To: {200601161533.k0GFXXfg018559-at-ns.microscopy.com} 5, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
The College of Microscopy will be offering a short course, Scanning Electron Microscopy, March 27-31, 2006, at our Westmont Facility. In addition to lectures, the course emphasizes hands-on training using five scanning electron microscopes and electron microprobe analyzers, and gives students the opportunity to work on their own samples. For further details and registration information, please follow the link below.
www.collegeofmicroscopy.com
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
==============================Original Headers============================== 8, 27 -- From eschumacher-at-mccrone.com Mon Jan 16 11:56:32 2006 8, 27 -- Received: from pgp.mccrone.com (McCrone-Associates-1051626.cust-rtr.ameritech.net [68.23.63.122]) 8, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0GHuW0p005961 8, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Jan 2006 11:56:32 -0600 8, 27 -- Received: from pgp.mccrone.com (localhost [127.0.0.1]) 8, 27 -- by pgp.mccrone.com (Postfix) with ESMTP id 797C11A800B 8, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Jan 2006 11:56:34 -0600 (CST) 8, 27 -- Received: from MCCRONEMSG.tmg.mccrone.com ([192.168.101.20]) 8, 27 -- by pgp.mccrone.com (PGP Universal service); 8, 27 -- Mon, 16 Jan 2006 11:56:34 -0600 8, 27 -- X-PGP-Universal: processed 8, 27 -- MIME-Version: 1.0 8, 27 -- Content-Type: text/plain; 8, 27 -- charset="US-ASCII" 8, 27 -- Content-class: urn:content-classes:message 8, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.6944.0 8, 27 -- Subject: Short Course Announcement: Scanning Electron Microscopy 8, 27 -- Date: Mon, 16 Jan 2006 11:56:21 -0600 8, 27 -- Message-ID: {2A27CE0EC2A5C748974EA513DFB285A74C310A-at-MCCRONEMSG.tmg.mccrone.com} 8, 27 -- X-MS-Has-Attach: 8, 27 -- X-MS-TNEF-Correlator: 8, 27 -- Thread-Topic: Short Course Announcement: Scanning Electron Microscopy 8, 27 -- thread-index: AcYaxijJDPWYg/gFR2u07q3UCQ75Vw== 8, 27 -- From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} 8, 27 -- To: {Microscopy-at-MSA.Microscopy.Com} 8, 27 -- Content-Transfer-Encoding: 8bit 8, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0GHuW0p005961 ==============================End of - Headers==============================
DISCLAIMER: South Bay Technology produces equipment and supplies as described below and, therefore, has a vested interest in promoting their use.
If you are mounting the sample in AcryliMet™ cold mount or something similar and can hold your sample by hand, then something like our Model 900 Grinder/Polisher would be ideal. A new Model 900 falls into about the same price range as the used Minimet you referred to. This system is often used for low volume polishing requirements - although usually for higher volume than just 1 sample per week. If you have unencapsulated samples or simply want more control over the polishing process, you can use the Model 900 together with something like our Tripod Polisher® Cross Section Polisher, our BiPod™ Cross Section Polisher or one of our other lapping and polishing fixtures. You can find information on the Model 900 Polisher at http://southbaytech.com/products_index.cfm?main_action=product_detail&ProductID=42.
If you do end up with the Minimet Polisher, I do have a large lot of Minimet Consumables available at a huge discount. If you have an interest in those, please let me know and I will send you the listing.
Best regards-
David
lemaster-at-ppg.com wrote:
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==============================Original Headers============================== 20, 21 -- From henriks-at-southbaytech.com Mon Jan 16 13:06:35 2006 20, 21 -- Received: from ylpvm25.prodigy.net (ylpvm25-ext.prodigy.net [207.115.57.56]) 20, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0GJ6ZDe016034 20, 21 -- for {microscopy-at-msa.microscopy.com} ; Mon, 16 Jan 2006 13:06:35 -0600 20, 21 -- Received: from southbaytech.com (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 20, 21 -- (authenticated bits=0) 20, 21 -- by ylpvm25.prodigy.net (smtpauth/dk/flock 8.13.4/8.13.4) with ESMTP id k0GJ68Ik027256; 20, 21 -- Mon, 16 Jan 2006 14:06:10 -0500 20, 21 -- Message-ID: {43CBEEC8.3020505-at-southbaytech.com} 20, 21 -- Date: Mon, 16 Jan 2006 11:06:48 -0800 20, 21 -- From: David Henriks {henriks-at-southbaytech.com} 20, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 20, 21 -- X-Accept-Language: en-us, en 20, 21 -- MIME-Version: 1.0 20, 21 -- To: lemaster-at-ppg.com 20, 21 -- CC: Microscopy Listerver {microscopy-at-msa.microscopy.com} 20, 21 -- Subject: Re: [Microscopy] SEM advice on low cost polisher 20, 21 -- References: {200601161548.k0GFmtuG021248-at-ns.microscopy.com} 20, 21 -- In-Reply-To: {200601161548.k0GFmtuG021248-at-ns.microscopy.com} 20, 21 -- Content-Type: text/plain; charset=windows-1252; format=flowed 20, 21 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I just returned from a trip out of the country, and found an inquiry about starting up ion pumps in among 1200 other assorted e-mail messages (mostly spam), and so I apologize for being so delayed in answering.
I don't know the exact details of the arrangement of the vacuum system on a JEOL 2010, but in general there should be no requirement for having cooling water running while starting up or running a sputter ion pump. In fact, one of the principle advantages of these pumps is the fact that they do not need cooling of any sort for normal operation.
What you would need to do is to set the valves in the vacuum system so that the backing pump evacuates the region of the vacuum system served by the ion pumps, then evacuate this region to a vacuum in the low 10-3 torr range. Then, follow the start-up procedure recommended for your ion pumps. You want to minimize the time the backing pumps are operated in the low end of this pressure range to reduce the possibility of contaminating the system by back streaming of oil from them, otherwise, there should be no problem starting up the ion pumps. As soon as they do start you want to close whatever valves are available to isolate the part of the system they serve from the part evacuated by the other pumps. Once the supply of cooling water is restored, use the diffusion pump to evacuate the rest of the system to an operating level, and then open the valves to the ion-pumped region and resume normal operation.
If you can get hold of a copy of my book on Vacuum Methods in Electron Microscopy (ISBN 1 85578 053 4) you can find the matter of backstreaming from rotary vane pumps discussed in Section 4.1.5 on p. 144. The general operating characteristics of sputter ion pumps is discussed in Sections 7.1.6, p 290 and 7.5, p. 322. I believe the operating procedure for the vacuum system described in Section 9.3 on p. 392 is very similar to that for the 2010.
Good luck, Wil Bigelow
-- 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============================== 7, 14 -- From bigelow-at-engin.umich.edu Mon Jan 16 14:34:37 2006 7, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 7, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0GKYZh8026672 7, 14 -- for {microscopy-at-microscopy.com} ; Mon, 16 Jan 2006 14:34:36 -0600 7, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 7, 14 -- by srvr22.engin.umich.edu (8.12.10/8.12.10) with ESMTP id k0GKYYGU013312 7, 14 -- for {microscopy-at-microscopy.com} ; Mon, 16 Jan 2006 15:34:34 -0500 (EST) 7, 14 -- Mime-Version: 1.0 7, 14 -- Message-Id: {p06210201bff1b09494cf-at-[141.212.131.221]} 7, 14 -- Date: Mon, 16 Jan 2006 15:34:33 -0500 7, 14 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 7, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 7, 14 -- Subject: RE:[Microscopy] Starting Ion Pumps without cooling water 7, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
I too am not aware of any water cooling for ion pumps. when first pumping, they do get warm. But their controller will shut them off for a cool down.
The main problem I have had in the past is getting them to start pumping. Initially, no pump current. This is fixed by heating the pump with a hair dryer. Then they fire and that is that. This is typical of the little 1-5L/m gun chamber pumps for LaB6. The larger 30L/m pumps don't seem to have this problem. This was with Varian pumps.
gary g.
At 12:53 PM 1/16/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Mon Jan 16 15:40:09 2006 11, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0GLe9od004433 11, 20 -- for {microscopy-at-microscopy.com} ; Mon, 16 Jan 2006 15:40:09 -0600 11, 20 -- Received: (qmail 5325 invoked from network); 16 Jan 2006 13:40:08 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 5322, pid: 5323, t: 0.1787s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp2 with SMTP; 16 Jan 2006 13:40:08 -0800 11, 20 -- Message-Id: {6.2.3.4.2.20060116133653.0206f088-at-mail.calweb.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 11, 20 -- Date: Mon, 16 Jan 2006 13:40:08 -0800 11, 20 -- To: bigelow-at-engin.umich.edu 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] RE: Starting Ion Pumps without cooling water 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200601162053.k0GKrXr6030172-at-ns.microscopy.com} 11, 20 -- References: {200601162053.k0GKrXr6030172-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I have a multi-function gauge on my T300 that reads from 0-1. It is used with a rotating switch to measure all the voltages etc in the system. When its at full vacuum it reads 0.42 and the manual says it should be 6v which I assume is 0.6. I was wondering if anyone out there knows what this translates to in terms of Torr? Please reply off list if this is not of general interest.
Thanks in advance!
Tom Kaye
==============================Original Headers============================== 5, 20 -- From tom-at-tomkaye.com Mon Jan 16 20:44:11 2006 5, 20 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0H2iAoG017269 5, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Jan 2006 20:44:10 -0600 5, 20 -- Received: from tkdell [67.165.188.91] by tomkaye.com with ESMTP 5, 20 -- (SMTPD32-8.14) id A9F8638023E; Mon, 16 Jan 2006 20:44:08 -0600 5, 20 -- From: "Tom" {tom-at-tomkaye.com} 5, 20 -- To: {Microscopy-at-microscopy.com} 5, 20 -- Subject: Question about vacuum gauge on JEOL T300 5, 20 -- Date: Mon, 16 Jan 2006 20:44:09 -0600 5, 20 -- Message-ID: {021901c61b0f$e4b081b0$030a1aac-at-tkdell} 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="iso-8859-1" 5, 20 -- Content-Transfer-Encoding: 7bit 5, 20 -- X-Priority: 3 (Normal) 5, 20 -- X-MSMail-Priority: Normal 5, 20 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2910.0) 5, 20 -- Importance: Normal 5, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
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Email: cumings-at-umd.edu Name: John Cumings
Organization: University of Maryland
Title-Subject: [Filtered] Job Opening
Question: Dear Microscopy Community,
I would like to bring your attention to an open position in electron microscopy as the lab manager of a newly-created facility here at the University of Maryland. The position is part of the growing nanoscience center on campus, the Maryland Center for Integrated Nano Science and Engineering (M-CINSE).
More information can be found at http://mse.umd.edu/dept/positions/LabManagerUniversityMicroscopyFacility.htm
and applications can be submitted at https://apra.umd.edu/search.jsp?ID=ENMA000003
Please note that the "best consideration" deadline of Jan. 15th has just passed, so please apply soon if you are interested.
Best regards,
-John Cumings
-- John Cumings cumings-at-umd.edu Assistant Professor Department of Materials Science and Engineering University of Maryland College Park, MD 20742-2115
office (301) 405-0789 (1246 Kim Building) fax (301) 314-8164 --
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Email: nrvrctr-at-adelphia.net Name: Ed Beckett
Organization: MRI Virginia
Title-Subject: [Filtered] SEM
Question: I am searching for a Test engineer in Chicago with experience in SEM as well as other electrical and test abilities. Salary is about $60K. Contact me if you are aware of anyone or self interested. Thanks Ed Beckett 540-980-3100
I am interested to study magnetic structure (type II magnetic contrast) by SEM. Our SEM equipped with a Four Quadrant Backscattered Electron Detector. I will appreciate if you have experience using this type of detector for magnetic domain structure to guide us with the recommended working parameters as combinations of quadrant setting, working distance, tilt and etc.
Thank you in advanced,
Yossi.
==============================Original Headers============================== 5, 30 -- From yezer-at-cc.hut.fi Tue Jan 17 02:22:24 2006 5, 30 -- Received: from smtp-3.hut.fi (smtp-3.hut.fi [130.233.228.93]) 5, 30 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0H8MN4R016682 5, 30 -- for {Microscopy-at-Microscopy.Com} ; Tue, 17 Jan 2006 02:22:24 -0600 5, 30 -- Received: from localhost (putosiko.hut.fi [130.233.228.114]) 5, 30 -- by smtp-3.hut.fi (8.12.10/8.12.10) with ESMTP id k0H8MM6h013963 5, 30 -- for {Microscopy-at-Microscopy.Com} ; Tue, 17 Jan 2006 10:22:22 +0200 5, 30 -- Received: from smtp-3.hut.fi ([130.233.228.93]) 5, 30 -- by localhost (putosiko.hut.fi [130.233.228.114]) (amavisd-new, port 10024) 5, 30 -- with LMTP id 22195-17-9 for {Microscopy-at-Microscopy.Com} ; 5, 30 -- Tue, 17 Jan 2006 10:22:22 +0200 (EET) 5, 30 -- Received: from baresi.hut.fi (baresi-m.hut.fi [130.233.228.121]) 5, 30 -- by smtp-3.hut.fi (8.12.10/8.12.10) with ESMTP id k0H8LuRu013837 5, 30 -- for {Microscopy-at-Microscopy.Com} ; Tue, 17 Jan 2006 10:21:56 +0200 5, 30 -- Received: (from apache-at-localhost) 5, 30 -- by baresi.hut.fi (8.12.10/8.12.6/Submit) id k0H8LtkJ029142 5, 30 -- for Microscopy-at-Microscopy.Com; Tue, 17 Jan 2006 10:21:55 +0200 5, 30 -- To: Microscopy-at-Microscopy.Com 5, 30 -- Subject: Magnetic contrast with 4QBSED 5, 30 -- Message-ID: {1137486115.43cca9239de7c-at-webmail1.hut.fi} 5, 30 -- Date: Tue, 17 Jan 2006 10:21:55 +0200 (EET) 5, 30 -- From: yezer-at-cc.hut.fi 5, 30 -- MIME-Version: 1.0 5, 30 -- Content-Type: text/plain; charset=ISO-8859-1 5, 30 -- Content-Transfer-Encoding: 8bit 5, 30 -- User-Agent: HUT webmail, IMP 2.2.6 5, 30 -- X-Authenticated-Sender: yezer-at-cc.hut.fi 5, 30 -- X-Originating-IP: 130.233.108.169 5, 30 -- X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on putosiko.hut.fi 5, 30 -- X-TKK-Virus-Scanned: by amavisd-new-2.1.2-hutcc at putosiko.hut.fi ==============================End of - Headers==============================
The Naval Research Laboratory has a current opening for a Postdoctoral Fellow in the Joining and Transformations Section. The ideal candidate would have a strong background in both electron microscopy and materials science. The successful candidate will study the transformations that occur during joining processes and relate the microstructural features to observed mechanical properties. Current research interests center on the precipitation, dynamic recrystallization, and texture evolution that occur during the friction stir welding process.
The Naval Research Laboratory has an excellent array of microscopy facilities available. There are three transmission electron microscopes (a Phillips CM-30 analytical TEM, a Hitachi H-9000 high-resolution TEM, and a new, state-of-the-art JEOL 2200 energy filtered STEM/TEM), two scanning electron microscopes (a Leo 1550 SEM with electron backscattered diffraction analysis and energy dispersive spectrometry and a Hitachi FEG-SEM), and a dual-beam focused ion beam with EBSD that are available for use by the successful candidate. Additional facilities that may be used include a quenching and deformation dilatometer, a Gleeble thermomechanical simulator, x-ray diffractometers, and an automated microhardness tester.
Please note that this position is only open to US citizens or Green Card holders.
Please contact me by e-mail (Fonda-at-anvil.nrl.navy.mil) or phone (202-767-2622) for further details about this research program.
Sinceerely,
Richard Fonda -- ________________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 ________________________________________________________________
==============================Original Headers============================== 6, 14 -- From fonda-at-anvil.nrl.navy.mil Tue Jan 17 06:21:09 2006 6, 14 -- Received: from anvil.nrl.navy.mil (anvil.nrl.navy.mil [132.250.127.188]) 6, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HCL8kO029509 6, 14 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 06:21:09 -0600 6, 14 -- Received: from [132.250.127.172] (rw-fonda.nrl.navy.mil [132.250.127.172]) 6, 14 -- by anvil.nrl.navy.mil (8.12.11/8.12.11) with ESMTP id k0HCL5eT022874 6, 14 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 07:21:06 -0500 (EST) 6, 14 -- Mime-Version: 1.0 6, 14 -- Message-Id: {p06230901bff2920cbcd0-at-[132.250.127.172]} 6, 14 -- Date: Tue, 17 Jan 2006 07:22:41 -0500 6, 14 -- To: Microscopy-at-microscopy.com 6, 14 -- From: "Richard W. Fonda" {fonda-at-anvil.nrl.navy.mil} 6, 14 -- Subject: Postdoctoral Fellowship opening at NRL 6, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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Email: lleroux-at-csir.co.za Name: Lukas le Roux
Organization: CSIR
Title-Subject: [Filtered] Edwards Sputter Coater
Question: I am trying to find an instruction manual for a model S150B Sputter Coater. Is there a website from where I can download it?
Is there a company that can provide room survey services in the Dallas, Texas area? Environmental testing for mechanical vibration and stray EMF parameters are desired.
Please respond directly off list.
Bob Roberts EM Lab Services, Inc. 449 NW 62nd St. Topeka, Kansas 66617-1780 785.246.1232 voice 785.246.0168 fax www.emlabservices.com
==============================Original Headers============================== 6, 23 -- From emlabservices-at-cox.net Tue Jan 17 07:35:50 2006 6, 23 -- Received: from centrmmtao06.cox.net (centrmmtao06.cox.net [70.168.83.78]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HDZm75007409 6, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 07:35:49 -0600 6, 23 -- Received: from EMLabServices ([24.255.213.54]) by centrmmtao06.cox.net 6, 23 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with SMTP 6, 23 -- id {20060117133551.RBDG4002.centrmmtao06.cox.net-at-EMLabServices} 6, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 08:35:51 -0500 6, 23 -- Message-ID: {008601c61b6a$f0ad2440$6400a8c0-at-EMLabServices} 6, 23 -- From: "EM Lab Services" {emlabservices-at-cox.net} 6, 23 -- To: {Microscopy-at-microscopy.com} 6, 23 -- Subject: TEM Room Survey Company 6, 23 -- Date: Tue, 17 Jan 2006 06:35:53 -0700 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- format=flowed; 6, 23 -- charset="iso-8859-1"; 6, 23 -- reply-type=original 6, 23 -- Content-Transfer-Encoding: 7bit 6, 23 -- X-Priority: 3 6, 23 -- X-MSMail-Priority: Normal 6, 23 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 6, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
"I am discussing proposals to install an imaging system here. It is one that can capture both FISH and brown/ H&E stains and then allow an operator, using selected software, to analyse them. The favoured system at present is the Applied Imaging one as it is now undergoing a major facelift.
I am just wondering if there are any other systems on the market right now that can load up 50 slides automatically overnight, scan & store so that an operator can then analyse the images captured the following morning. None of the other major suppliers here in the UK, such as ChromaVision, Meta-systems & Aperio can offer both overnight loading and scanning of both FISH and brown/ blue & pink stains at present."
Gareth Morgan MPhil MSc FIBMS, Department of Histo/cytopathology, Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
==============================Original Headers============================== 14, 18 -- From Gareth.Morgan-at-ki.se Tue Jan 17 10:10:41 2006 14, 18 -- Received: from humle.it.ki.se (humle.it.ki.se [130.237.101.252]) 14, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HGAeFM027877 14, 18 -- for {microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 10:10:41 -0600 14, 18 -- Received: from lgtestdator.ki.se (hsg01.hs.se [193.10.76.5]) 14, 18 -- by humle.it.ki.se (8.13.1/8.13.1) with ESMTP id k0HGAeo4003655 14, 18 -- for {microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 17:10:40 +0100 (MET) 14, 18 -- Message-Id: {7.0.0.16.0.20060117171215.020160a8-at-ki.se} 14, 18 -- Message-Id: {7.0.0.16.0.20060116170224.020ac968-at-ki.se} 14, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.0.16 14, 18 -- Date: Tue, 17 Jan 2006 17:12:17 +0100 14, 18 -- To: microscopy-at-microscopy.com 14, 18 -- From: Gareth Morgan {Gareth.Morgan-at-ki.se} 14, 18 -- Subject: Image analysis 14, 18 -- Mime-Version: 1.0 14, 18 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 14, 18 -- Content-Transfer-Encoding: 8bit 14, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0HGAeFM027877 ==============================End of - Headers==============================
We're having trouble with bright spots appearing on our tissue when viewing our grids in our new TEM. These spots may be described as bleaching, etching, burning or clearing. The bright spots appears after using high magnification to view an area or to focus. When we go back to a low magnification, we see the bright area.
This started happening with the installation of a new TEM. We haven't changed processing procedures and we are experienced instrument operators. The service engineer has determined that the TEM is working properly. The problem is not due to prolonged exposure to the electron beam. The spots occur within seconds. The filament is positioned correctly. Often there is variability in the intensity of the spotting even on the same grid. In other words, sometime the spot is very apparent and distinctive and other times it's more diffuse. We've tried using liquid nitrogen to improve the vacuum and adding a second 100% resin infiltration step to ensure removal of water from the specimen. The problem occurs with cells and tissues embedded in epoxy. The vendor has suggested working at 120kV and we're starting to do that now. It doesn't entirely correct the problem, but the spots are less focal.
Has anyone worked with a particular epoxy that stands up well or better than others under the electron beam? You can reply off list. Has anyone had this problem and been able find a solution?
Thank you in advance for your time
Marcia
Marcia Pitzenberger Pathology Laboratories Merck & Co., Inc. P.O. Box 4, WP45-104 West Point, PA 19486-0004 Tel 215 652-9767 Fax 215 652-7758 marcia_pitzenberger-at-merck.com
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==============================Original Headers============================== 13, 24 -- From marcia_pitzenberger-at-merck.com Tue Jan 17 11:12:37 2006 13, 24 -- Received: from usryim09.merck.com (taz.merck.com [155.91.6.20]) 13, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HHCaxL008406 13, 24 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Jan 2006 11:12:36 -0600 13, 24 -- Received: from 155.91.2.6 by usryim09.merck.com with ESMTP (SMTP Relay); 13, 24 -- Tue, 17 Jan 2006 12:12:27 -0500 13, 24 -- X-Server-Uuid: 078441B8-8B94-4128-83AA-0F28411313D9 13, 24 -- Received: from 54.3.102.166 by USRYTW32.merck.com with ESMTP (Tumbleweed 13, 24 -- Email Firewall SMTP Relay (Email Firewall v6.1.1)); Tue, 17 Jan 2006 13, 24 -- 12:12:16 -0500 13, 24 -- X-Server-Uuid: 05354929-4888-4C60-B5F7-73C306CFF21D 13, 24 -- Received: by usrygw30.merck.com with Internet Mail Service (5.5.2658.27) 13, 24 -- id {C9J1FVY2} ; Tue, 17 Jan 2006 12:12:16 -0500 13, 24 -- Message-ID: {4F1089C79438304CB5A97E982A11AC1F05AF89-at-usctmx1118.merck.com} 13, 24 -- From: "Pitzenberger, Marcia H." {marcia_pitzenberger-at-merck.com} 13, 24 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-msa.microscopy.com} 13, 24 -- Subject: TEM - Electron Beam Clearing 13, 24 -- Date: Tue, 17 Jan 2006 12:12:07 -0500 13, 24 -- MIME-Version: 1.0 13, 24 -- X-Mailer: Internet Mail Service (5.5.2658.27) 13, 24 -- X-WSS-ID: 6FD3FAFA1X466977-01-01 13, 24 -- X-WSS-ID: 6FD3FAF12C419349-01-01 13, 24 -- Content-Type: text/plain 13, 24 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
On Jan 14, 2006, at 5:35 AM, mswift-at-bunham.org wrote:
} Question: We have a Tecnai 12 from FEI, and we would like to know: How } do we ensure that in Low Dose our Focus Substates 1 and 2 are on the } tilt axis when Alpha angle is not 0 degrees? } Dear Mark, If you set the focus angles for the substates to 0 and 180 degrees, the displacements will be along the tilt axis. You can verify this by burning a small hole in the ice on a cryogrid in each of the focus substates, then going to a lower mag, where the holes are both visible in the exposure state, and tilting the grid. 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 Jan 17 12:14:06 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HIE5Ii019810 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Jan 2006 12:14:05 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 9B83B109E08 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Jan 2006 10:14:03 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id B68A633C9D 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Jan 2006 10:14:02 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200601141335.k0EDZ0gB029586-at-ns.microscopy.com} 4, 22 -- References: {200601141335.k0EDZ0gB029586-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {b7eeb9658bb9479e350b822656f74027-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] AskAMicroscopist: Tecnai 12 Focus Substates 4, 22 -- Date: Tue, 17 Jan 2006 10:20:58 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
I speak from a position of ignorance of your type of specimen, however I have picked up on 'This started happening with the installation of a new TEM'.
Are you using a much higher beam current at high mags than previously? Modern TEMs have much better condenser optics than the older ranges available in the 70s and 80s with less use of apertures to reduce spot size and better use of lenses. Have you changed from tungsten to LaB6 or FEG?
These changes can result in more electrons getting to the specimen. Can you compare the beam current that you use at high mag on the two TEMs? If you know typical exposure times of the two instruments do you have to spread the beam more on the new instument to get the same exposure time? Try using a smaller spot size or smaller condenser aperture and see if the effect improves.
Good luck, Ron
In message {200601171752.k0HHqH72018071-at-ns.microscopy.com} marcia_pitzenberger-at-merck.com writes: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } TEM User's - } } } We're having trouble with bright spots appearing on our tissue when viewing } our grids in our new TEM. These spots may be described as bleaching, } etching, burning or clearing. The bright spots appears after using high } magnification to view an area or to focus. When we go back to a low } magnification, we see the bright area. } } This started happening with the installation of a new TEM. We haven't } changed processing procedures and we are experienced instrument operators. } The service engineer has determined that the TEM is working properly. The } problem is not due to prolonged exposure to the electron beam. The spots } occur within seconds. The filament is positioned correctly. Often there is } variability in the intensity of the spotting even on the same grid. In } other words, sometime the spot is very apparent and distinctive and other } times it's more diffuse. We've tried using liquid nitrogen to improve the } vacuum and adding a second 100% resin infiltration step to ensure removal of } water from the specimen. The problem occurs with cells and tissues } embedded in epoxy. The vendor has suggested working at 120kV and we're } starting to do that now. It doesn't entirely correct the problem, but the } spots are less focal. } } Has anyone worked with a particular epoxy that stands up well or better than } others under the electron beam? You can reply off list. Has anyone had } this problem and been able find a solution? } } Thank you in advance for your time } } Marcia } } } } Marcia Pitzenberger } Pathology Laboratories } Merck & Co., Inc. } P.O. Box 4, WP45-104 } West Point, PA 19486-0004 } Tel 215 652-9767 } Fax 215 652-7758 } marcia_pitzenberger-at-merck.com } } } } } } ----------------------------------------------------------------------------- - } Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. } ----------------------------------------------------------------------------- - } } ==============================Original Headers============================== } 13, 24 -- From marcia_pitzenberger-at-merck.com Tue Jan 17 11:12:37 2006 } 13, 24 -- Received: from usryim09.merck.com (taz.merck.com [155.91.6.20]) } 13, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HHCaxL008406 } 13, 24 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Jan 2006 11:12:36 -0600 } 13, 24 -- Received: from 155.91.2.6 by usryim09.merck.com with ESMTP (SMTP Relay); } 13, 24 -- Tue, 17 Jan 2006 12:12:27 -0500 } 13, 24 -- X-Server-Uuid: 078441B8-8B94-4128-83AA-0F28411313D9 } 13, 24 -- Received: from 54.3.102.166 by USRYTW32.merck.com with ESMTP (Tumbleweed } 13, 24 -- Email Firewall SMTP Relay (Email Firewall v6.1.1)); Tue, 17 Jan 2006 } 13, 24 -- 12:12:16 -0500 } 13, 24 -- X-Server-Uuid: 05354929-4888-4C60-B5F7-73C306CFF21D } 13, 24 -- Received: by usrygw30.merck.com with Internet Mail Service (5.5.2658.27) } 13, 24 -- id {C9J1FVY2} ; Tue, 17 Jan 2006 12:12:16 -0500 } 13, 24 -- Message-ID: {4F1089C79438304CB5A97E982A11AC1F05AF89-at-usctmx1118.merck.com} } 13, 24 -- From: "Pitzenberger, Marcia H." {marcia_pitzenberger-at-merck.com} } 13, 24 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-msa.microscopy.com} } 13, 24 -- Subject: TEM - Electron Beam Clearing } 13, 24 -- Date: Tue, 17 Jan 2006 12:12:07 -0500 } 13, 24 -- MIME-Version: 1.0 } 13, 24 -- X-Mailer: Internet Mail Service (5.5.2658.27) } 13, 24 -- X-WSS-ID: 6FD3FAFA1X466977-01-01 } 13, 24 -- X-WSS-ID: 6FD3FAF12C419349-01-01 } 13, 24 -- Content-Type: text/plain } 13, 24 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
-- Mr. Ron Doole Department of Materials Senior Instrumentation Engineer. University of Oxford. Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
==============================Original Headers============================== 7, 27 -- From ron.doole-at-materials.ox.ac.uk Tue Jan 17 12:26:00 2006 7, 27 -- Received: from relay9.mail.ox.ac.uk (relay9.mail.ox.ac.uk [163.1.2.169]) 7, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HIPxqX022315 7, 27 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Jan 2006 12:25:59 -0600 7, 27 -- Received: from smtp1.herald.ox.ac.uk ([163.1.0.247]) 7, 27 -- by relay9.mail.ox.ac.uk with esmtp (Exim 4.54) 7, 27 -- id 1EyvWo-0007nk-WD 7, 27 -- for microscopy-at-msa.microscopy.com; Tue, 17 Jan 2006 18:25:59 +0000 7, 27 -- Received: from webmail222.herald.ox.ac.uk ([163.1.0.222]) 7, 27 -- by smtp1.herald.ox.ac.uk with esmtp (Exim 3.36 #1) 7, 27 -- id 1EyvWo-0004Bf-01 7, 27 -- for microscopy-at-msa.microscopy.com; Tue, 17 Jan 2006 18:25:58 +0000 7, 27 -- Received: by webmail222.herald.ox.ac.uk (Postfix, from userid 101) 7, 27 -- id AD78A2A0B8; Tue, 17 Jan 2006 18:25:58 +0000 (GMT) 7, 27 -- Content-Type: text/plain 7, 27 -- Content-Disposition: inline 7, 27 -- Content-Transfer-Encoding: binary 7, 27 -- MIME-Version: 1.0 7, 27 -- X-Mailer: MIME-tools 5.411 (Entity 5.404) 7, 27 -- From: Ron Doole {ron.doole-at-materials.ox.ac.uk} 7, 27 -- Date: Tue, 17 Jan 2006 18:25:58 +0000 (GMT) 7, 27 -- In-Reply-To: {200601171752.k0HHqH72018071-at-ns.microscopy.com} 7, 27 -- To: microscopy-at-msa.microscopy.com 7, 27 -- Subject: Re: [Microscopy] TEM - Electron Beam Clearing 7, 27 -- X-Webmail-Originating-Ip: 192.76.27.49 7, 27 -- X-Webmail-Sender: rdoole 7, 27 -- Message-Id: {20060117182558.AD78A2A0B8-at-webmail222.herald.ox.ac.uk} ==============================End of - Headers==============================
Gary; Hitting ion pumps with a hammer also gets them started. The hammer is also very good at knocking loose whiskers that form in the ion pump.
John Mardinly Intel
The opinion of this author does not necessarily represent the opinion of Intel Corporation.
-----Original Message----- X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com] Sent: Monday, January 16, 2006 1:41 PM To: Mardinly, John
I too am not aware of any water cooling for ion pumps. when first pumping, they do get warm. But their controller will shut them off for a cool down.
The main problem I have had in the past is getting them to start pumping. Initially, no pump current. This is fixed by heating the pump with a hair dryer. Then they fire and that is that. This is typical of the little 1-5L/m gun chamber pumps for LaB6. The larger 30L/m pumps don't seem to have this problem. This was with Varian pumps.
gary g.
At 12:53 PM 1/16/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Mon Jan 16 15:40:09 2006 11, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0GLe9od004433 11, 20 -- for {microscopy-at-microscopy.com} ; Mon, 16 Jan 2006 15:40:09 -0600 11, 20 -- Received: (qmail 5325 invoked from network); 16 Jan 2006 13:40:08 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 5322, pid: 5323, t: 0.1787s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp2 with SMTP; 16 Jan 2006 13:40:08 -0800 11, 20 -- Message-Id: {6.2.3.4.2.20060116133653.0206f088-at-mail.calweb.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 11, 20 -- Date: Mon, 16 Jan 2006 13:40:08 -0800 11, 20 -- To: bigelow-at-engin.umich.edu 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] RE: Starting Ion Pumps without cooling water 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200601162053.k0GKrXr6030172-at-ns.microscopy.com} 11, 20 -- References: {200601162053.k0GKrXr6030172-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 20, 34 -- From john.mardinly-at-intel.com Tue Jan 17 15:16:13 2006 20, 34 -- Received: from scsfmr003.sc.intel.com (fmr23.intel.com [143.183.121.15]) 20, 34 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HLGCFX009840 20, 34 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Jan 2006 15:16:13 -0600 20, 34 -- Received: from scsfmr101.sc.intel.com (scsfmr101.sc.intel.com [10.3.253.10]) 20, 34 -- by scsfmr003.sc.intel.com (8.12.10/8.12.10/d: major-outer.mc,v 1.1 2004/09/17 17:50:56 root Exp $) with ESMTP id k0HLGCK7006008; 20, 34 -- Tue, 17 Jan 2006 21:16:12 GMT 20, 34 -- Received: from scsmsxvs041.sc.intel.com (scsmsxvs041.sc.intel.com [10.3.90.10]) 20, 34 -- by scsfmr101.sc.intel.com (8.12.10/8.12.10/d: major-inner.mc,v 1.2 2004/09/17 18:05:01 root Exp $) with SMTP id k0HLC3jR001619; 20, 34 -- Tue, 17 Jan 2006 21:12:06 GMT 20, 34 -- Received: from scsmsx331.amr.corp.intel.com ([10.3.90.4]) 20, 34 -- by scsmsxvs041.sc.intel.com (SAVSMTP 3.1.7.47) with SMTP id M2006011713160707553 20, 34 -- ; Tue, 17 Jan 2006 13:16:07 -0800 20, 34 -- Received: from scsmsx403.amr.corp.intel.com ([10.3.90.18]) by scsmsx331.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 20, 34 -- Tue, 17 Jan 2006 13:16:07 -0800 20, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 20, 34 -- Content-class: urn:content-classes:message 20, 34 -- MIME-Version: 1.0 20, 34 -- Content-Type: text/plain; 20, 34 -- charset="us-ascii" 20, 34 -- Subject: RE: [Microscopy] Starting Ion Pumps without cooling water 20, 34 -- Date: Tue, 17 Jan 2006 13:16:03 -0800 20, 34 -- Message-ID: {9E1ED6A623D8CB44B4866ACF20ECDB2B08CC4267-at-scsmsx403.amr.corp.intel.com} 20, 34 -- X-MS-Has-Attach: 20, 34 -- X-MS-TNEF-Correlator: 20, 34 -- Thread-Topic: [Microscopy] Starting Ion Pumps without cooling water 20, 34 -- Thread-Index: AcYa5dJSmizC6M7cSkqL66kBhLViXgAxSKoQ 20, 34 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 20, 34 -- To: {gary-at-gaugler.com} 20, 34 -- Cc: {Microscopy-at-MSA.Microscopy.Com} 20, 34 -- X-OriginalArrivalTime: 17 Jan 2006 21:16:07.0634 (UTC) FILETIME=[3B630320:01C61BAB] 20, 34 -- X-Scanned-By: MIMEDefang 2.52 on 10.3.253.10 20, 34 -- Content-Transfer-Encoding: 8bit 20, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0HLGCFX009840 ==============================End of - Headers==============================
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Several glass plates ~5 mm thick and 15 cm square. Polishing clothes stuck to the glass plates. Glass plates inside large tray. Water spray to lubricate. Cost is almost nothing apart from the time. Depends how large the sample is and what finish you are working from. Fine grinding can be done by using aumina directly onto the glass. It's the way I used to prepare 3 mm steel discs for TEM, before a final electropolish. A 3 mm stainless steel disc takes ~15 mins to polish by hand from the alumina grinding.
-- Larry Stoter
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
==============================Original Headers============================== 4, 16 -- From larry-at-cymru.freewire.co.uk Tue Jan 17 15:45:45 2006 4, 16 -- Received: from get.freewire.net (get.freewire.net [195.184.229.250]) 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0HLjhPu015336 4, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Jan 2006 15:45:44 -0600 4, 16 -- Received: from [217.154.248.196] (th1dc-217-154-248-196.dial.mistral.co.uk [217.154.248.196]) 4, 16 -- by get.freewire.net (8.11.6/8.11.6) with ESMTP id k0HLjdC13695; 4, 16 -- Tue, 17 Jan 2006 21:45:40 GMT 4, 16 -- Mime-Version: 1.0 4, 16 -- Message-Id: {p06210200bff31079bb79-at-[217.154.248.193]} 4, 16 -- In-Reply-To: {200601161546.k0GFkBfX020754-at-ns.microscopy.com} 4, 16 -- References: {200601161546.k0GFkBfX020754-at-ns.microscopy.com} 4, 16 -- Date: Tue, 17 Jan 2006 21:33:35 +0000 4, 16 -- To: lemaster-at-ppg.com, Microscopy-at-MSA.Microscopy.Com 4, 16 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 4, 16 -- Subject: Re: [Microscopy] SEM advice on low cost polisher 4, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Oh yeah. I forgot to mention that trick too. I usually use the handle end of a screwdriver. This also works for cold cathode sensors too.
gary g.
At 01:33 PM 1/17/2006, you wrote:
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==============================Original Headers============================== 8, 20 -- From gary-at-gaugler.com Tue Jan 17 17:13:09 2006 8, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 8, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0HND8OF000462 8, 20 -- for {microscopy-at-microscopy.com} ; Tue, 17 Jan 2006 17:13:08 -0600 8, 20 -- Received: (qmail 2929 invoked from network); 17 Jan 2006 15:13:06 -0800 8, 20 -- Received: by simscan 1.1.0 ppid: 2926, pid: 2927, t: 0.2584s 8, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 8, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 8, 20 -- by qsmtp1 with SMTP; 17 Jan 2006 15:13:06 -0800 8, 20 -- Message-Id: {6.2.3.4.2.20060117151146.01febf50-at-mail.calweb.com} 8, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 8, 20 -- Date: Tue, 17 Jan 2006 15:13:07 -0800 8, 20 -- To: john.mardinly-at-intel.com 8, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 8, 20 -- Subject: Re: [Microscopy] RE: Starting Ion Pumps without cooling water 8, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 8, 20 -- In-Reply-To: {200601172133.k0HLXwjI013245-at-ns.microscopy.com} 8, 20 -- References: {200601172133.k0HLXwjI013245-at-ns.microscopy.com} 8, 20 -- Mime-Version: 1.0 8, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
With some degree of simplification- low signal electronic imaging is a race between noise accumulation and signal accumulation.
Some of the noise sources in the CCD are:
a) thermal noise (reduced by factor of 2 for every 6 to 7 degrees C temperature drop for silicone CCD), mostly correctable by dark frame subtraction; b) readout (pixel clock switching) noise also know as bias noise, somewhat reduced by cooling, correctable by bias frame subtraction; c) random (shot) noise - a random part of thermal noise (a) which can not be corrected by dark frame subtraction- this noise gets worse with pixels size shrinking (the larger the pixels the less random this value is)- a crude comparison is a mechanical vibration dumping by increasing weight (anti-vibration platform).
Assuming your signal source is set (optics, specimen, illumination):
a) the colder the sensor- the better S/N ratio is- noise accumulation slows down, while signal stays the same; b) with 30 deg. C difference noise will be different by factor of 32 (30deg. = 6deg.*5; noise drops twice per every 6 degrees, thus 2 power 5 = 32 which is a dramatic noise reduction, but noticeable only if light is dim and exposures are long. c) the larger the pixels- the better S/N ratio is- at the expense of spatial resolution- it could pay to have more smaller pixels and use binning instead when needed.
Type of sensor:
a) definitely CCD over CMOS; b) probably full frame CCD over interline CCD, (full frame has much better S/N ratio for a given pixel size), but interline CCD could be more convenient due to potentially faster frame rate, exposure time allowing; c) if exposure time must be long, interline CCD looses it's potential speed advantage, and it is always noisier than full frame one; d) interline CCD has typically 50% of it's light receiving area occupied by transfer gates, meaning that only half of it is light sensitive- this can be improved by micro lens array - most interline CCDs are available in such configuration), but sensitive area of the pixel is still half of what it could be in full frame CCD with same pixel size, which means higher noise .
Interline CCDs can use frame integration with less or no cooling and achieve decent noise reduction, but this is always inferior to deeply cooled full frame CCD in single frame (long exposure) integration mode, with respect to S/N.
I deliberately used S/N (signal to noise) ratio instead just "noise", because it is precisely ratio that counts. If signal is strong (bright light), then it accumulates much faster than noise. Then exposures are short, and no CCD cooling needed. There is no straight answer without knowing the numbers for a given application.
A specific application could be best served by either a CMOS, or a full frame CCD, or interline CCD sensor, cooled or not- many more variables must be taken into account.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: {phillipst-at-missouri.edu} To: {vitalylazar-at-att.net} Sent: Friday, January 13, 2006 12:54 PM
Overview: Schafer Vallecitos Laboratory is seeking a Senior Engineer /Scientist to add to our mass spectrometry group. SVL performs materials characterization and related analytical services on commercial and government contracts. The activities of the group include chemical and elemental analysis of materials on a production basis, maintenance of several mass spectrometers and ancillary equipment, development of new or improved MS analysis techniques, and quality assurance of analytical data. Schafer is a technically strong firm with a reputation for quality and integrity. Schafer's reputation is a direct result of our dedicated, motivated, talented, and creative staff that is responsible for developing our outstanding business relationships with our customers. Our technical capabilities are vast and growing to provide innovations for the future.
Responsibilities: This position requires an individual who has proven technical abilities to function as an instrument designer, with emphasis on ion optics, electronics, and computer automation. Commercial design experience would be a plus, as the instrument should be maintainable and replicable, not a one-of-a-kind that requires a large staff to maintain. The ideal candidate must have a strong background in materials science or engineering, or a related field of physics, geology, chemistry, or metrology. Experience in design and operation of mass spectrometers, particularly TIMS, and other analytical instruments, is highly recommended. The candidate should have competence in vacuum technology, electronics, mechanical design, ion optics, and project administration.
Qualifications: Experience with analytical instrument control computer hardware and software is required. The candidate must be able to operate independently with occasional supervision.
Other qualifications include:
. Bachelor's degree in physical science or engineering. . Minimum 10 years of technical experience. . Demonstrated ability to solve technical problems. . Expertise in data evaluation and quality control. . Proven ability to write clear scientific reports and proposals. . Must be a US citizen with the ability to obtain government security clearance.
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There are two systems I know of that he may have overlooked:
One is the (dot)Scan, by Soft Imaging Systems The second is the Mirax system, by Carl Zeiss
I know they can physcially do what you require (overnight, automated loading and unloading), but I don't know their abilities in terms of stains.
Hope that helps a little bit! -Chris
--------------------------------- Christopher Hayden SPA Novartis Pharmaceuticals
Hi
Can anyone out there help a colleague?
"I am discussing proposals to install an imaging=20 system here. It is one that can capture both FISH=20 and brown/ H&E stains and then allow an operator,=20 using selected software, to analyse them. The=20 favoured system at present is the Applied Imaging=20 one as it is now undergoing a major facelift.
I am just wondering if there are any other=20 systems on the market right now that can load=20 up 50 slides automatically overnight, scan &=20 store so that an operator can then analyse the=20 images captured the following morning. None of=20 the other major suppliers here in the UK, such as=20 ChromaVision, Meta-systems & Aperio can offer=20 both overnight loading and scanning of both FISH=20 and brown/ blue & pink stains at present."
Dear Yossi, I have not seen a reply to your inquiry, so I will tell you what I know, which isn't much. Of all of the mechanisms for contrast in the back-scattered-electron (BSE) detector, the magnetic domain is the lowest. First is atomic number, second is channeling contrast and third is magnetic domain. This means you must eliminate all others to see the magnetic domain. The sample must be single phase, preferably single crystal and flat, parallel to the BSE detector, well-polished and, perhaps, electropolished. I have not done this, only seen talks about it. It takes a lot of beam current, an optimum working distance for the BSE (about 20 to 15 mm), all quadrants on and contrast on the BSE amplifier as high as practical. And a lot of patience. Good luck. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- X-from: {yezer-at-cc.hut.fi} To: {mager-at-interchange.ubc.ca} Sent: Tuesday, January 17, 2006 1:01 AM
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Email: bucana-at-audumla.mdacc.tmc.edu Name: Corazon D. Bucana
Organization: UT MD Anderson Cancer Center
Title-Subject: [Filtered] Separation of resin from glass tissue culture chamber slides
Question: We have emdedded tissue culture cells grown on tissue culture glass chambered slides using Polybed Epon 812, polymerized at 60C for 2 days. We tried to separate the glass by using liquid nitrogen followed by warm water bath but the glass did not separate from the polymerized resin. Can anyone suggest an alternative procedure to remove the glass from the block? Thanks.
We grow the cells on a coverslip (round coverslips are better and they will fit into the chamber) and remove them by hydrofluoric acid afterwards. Works very well, although the HF is not very pleasant to work with.
Good luck,
M.
-- Michael Jarnik, Ph.D. Electron Microscope Facility Fox Chase Cancer Center 7701 Burholme Ave. Philadelphia, PA 19111 Tel. 215-728-5675 Fax 215-728-2412
Name: Corazon D. Bucana
Organization: UT MD Anderson Cancer Center
Title-Subject: Separation of resin from glass tissue culture chamber slides
Question: We have emdedded tissue culture cells grown on tissue culture glass chambered slides using Polybed Epon 812, polymerized at 60C for 2 days. We tried to separate the glass by using liquid nitrogen followed by warm water bath but the glass did not separate from the polymerized resin. Can anyone suggest an alternative procedure to remove the glass from the block? Thanks.
==============================Original Headers============================== 13, 19 -- From M_Jarnik-at-fccc.edu Thu Jan 19 19:23:54 2006 13, 19 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 13, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0K1Nr1r023799 13, 19 -- for {microscopy-at-microscopy.com} ; Thu, 19 Jan 2006 19:23:53 -0600 13, 19 -- Received: from fccc.edu (emf4.fccc.edu [131.249.9.170]) 13, 19 -- by azah.fccc.edu (8.12.11/8.12.11) with ESMTP id k0K1Nrxr008126 13, 19 -- for {microscopy-at-microscopy.com} ; Thu, 19 Jan 2006 20:23:53 -0500 (EST) 13, 19 -- Message-ID: {43D03BA9.5060405-at-fccc.edu} 13, 19 -- Date: Thu, 19 Jan 2006 20:23:53 -0500 13, 19 -- From: Michal Jarnik {M_Jarnik-at-fccc.edu} 13, 19 -- Reply-To: M_Jarnik-at-fccc.edu 13, 19 -- Organization: Fox Chase Cancer Center 13, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 13, 19 -- X-Accept-Language: en,cs 13, 19 -- MIME-Version: 1.0 13, 19 -- To: microscopy-at-microscopy.com 13, 19 -- Subject: RE: Separation of resin from glass tissue culture chamber slides 13, 19 -- Content-Type: text/plain; charset=us-ascii; format=flowed 13, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hi. We too use concentrated hydrofluoric acid to dissolve glass coverslips. If you use plastic tripour beakers, plastic forceps, and plenty of rinse water, it doesn't cause problems. Since slides are thicker than coverslips, they may need may need longer time in the acid. Coverslips usually dissolve in 20 mins if all the edges are free of resin. To help the acid get at the free edges, file down the edges of the embedded slides using a metal file. Then immerse in acid under the fume hood in a plastic beaker. Check periodically by dipping in water and looking at the surface( wear nitrile gloves and work under the hood) The surface should be smooth and shiny with no patches of undissolved glass( looks like an iceberg melting). Rinse well under running water and dry in the embedding oven. Good luck!
JoAnn Buchanan Stanford University school of Medicine Stanford, CA 94305
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Title-Subject: [Filtered] Immunocytochemistry for TEM
Question: Greetings ListServers,
Does anyone have a general protocol for labelling anti-lucifer yellow using protein-A gold as a bridging antibody in the Locust (Orthopteran) brain? Should I try to immunogold pre-embedding, using a detergent such as Saponin? Should I dissect out the brain before fixation? I am hoping to use the immunogold labelling to help me visualize neuronal connections. Any advice is greatly appreciated.
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Title-Subject: [Filtered] liposomes analyzed by Freeze fracture SEM
Question: Hello,
I work for a company that is interested in getting some liposomes analyzed by Freeze fracture SEM or another SEM type technique. If there is any way you could put me into contact with some of your members who do this type of work or to post my request on a list serve, etc. it would be extremely helpful.
Thank you,
Michael Beverly Sirna Therapeutics, Inc. 303-546-8190
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Title-Subject: [Filtered] NANOBIO summer school in Cargese France
Question: Please find enclosed a flyer on the NANOBIO summer school in Cargese, Corsica, France from July 17th till 29th, 2006.
Watching single biological molecules at work is now possible and the unravelled features of the molecular machinery at the nanoscale are currently changing our view of the cell. Though, bridging the gap between nano- and micro-world is not easy: how to relate single molecule events to biological functions at the cell level? how do molecular structures assemble, coordinate and integrate in the wholeÖ? Thanks to physicists and biologists, we will address these questions and try to give some answers. Speakers will present recent concepts and methods in physics and biology dedicated to single molecule observation and description of the cell at the sub-micron level. Particular emphasis will be given to the optical methods, which allow the observation of live cells in physiological conditions.
This school is part of the Nanosciencestech Marie Curie Series of Events: http://www.france-optique.org/Nanosciencestech.html
The students, including PhDs, post-docs, and young researchers, will be encouraged to present their own work (also in adjacent fields) during poster sessions.
A European and more generally an international audience is targeted, and all lectures in the summer school will be given in English.
Important dates: Pre-registration: Application deadline 28 february 2006 Selected participant will be informed before the end of march
Registration and paying attendees deadline: 30 April 2006
All correspondence should be sent to
SociČtČ FranÁaise d'Optique Nanobio summer school c/o FranÁoise CHAVEL Centre Scientifique d'Orsay - B’t. 503 91403 ORSAY cedex France
next time, you should only polymerize for about 7-8 hours before popping the cells off. I then usually re-embed the pieces that come off and polymerize using standard times.
At 06:43 PM 01/19/06, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Greetings ListServers, does anybody have any experience using TUNEL assay on plants? I use the kit from Roche, nucleotides dyed with TMR, and have a problem that all the seccions (free-hand,fixed,penetrated with Triton and pectinase/cellulase solution), even those treated just with labeling solution without enzymes, are TUNEL positive. Do you know any reasonable explanation how could the nucleotides bind on the nucleus (and slightly on plasmatic membrane) without enzymes added?
Best regards,
Zuzana Lenochova Charles University Prague
==============================Original Headers============================== 7, 21 -- From lenoska1-at-seznam.cz Fri Jan 20 09:10:21 2006 7, 21 -- Received: from mx1.seznam.cz (mx1.seznam.cz [212.80.76.26]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0KFAIIa017250 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Jan 2006 09:10:19 -0600 7, 21 -- Received: (qmail 21368 invoked by uid 0); 20 Jan 2006 15:10:18 -0000 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- Date: Fri, 20 Jan 2006 16:10:16 +0100 (CET) 7, 21 -- Reply-To: =?iso-8859-2?Q?Zuzana=20Lenochov=E1?= {lenoska1-at-seznam.cz} 7, 21 -- From: =?iso-8859-2?Q?Zuzana=20Lenochov=E1?= {lenoska1-at-seznam.cz} 7, 21 -- Received: from [195.113.47.73] 7, 21 -- by email.seznam.cz with HTTP 7, 21 -- for lenoska1-at-seznam.cz; 7, 21 -- Fri, 20 Jan 2006 16:02:18 +0100 (CET) 7, 21 -- Subject: =?us-ascii?Q?fluorescence=20microscopy?= 7, 21 -- Mime-Version: 1.0 7, 21 -- Message-Id: {4290.7421-27152-1432019944-1137769816-at-seznam.cz} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- X-Abuse: helpdesk-at-seznam.cz 7, 21 -- X-Seznam-User: lenoska1-at-seznam.cz ==============================End of - Headers==============================
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I've never done this and I'm trying to remember an illustration from an old JEOL SEM brochure, which described an attachment for magnetic domain imaging.
From what I recall, the experimental setup was similar to that used now for EBSD. Sample at a high tilt with a detector to one side. In this case, a BSE detector. The idea was that the weak effect of the magnetic domains caused a small divergence of BS electrons. By positioning the detector a long way from the sample and at a high angle from the incident beam, the divergence was amplified. -- Larry Stoter
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Even though I am very experienced in Electron Microscopy, there are times when I cut LM sections that are extremely wrinkled, and they look like cracked glass. I normally don't experience this, but when it happens, even considering every possible variable, I'm still not exactly sure what might be at work here to give this sort of result.
I'm looking for ideas here as to what might be causing this effect. What do people think?
Garry Burgess Charge Technologist - Electron Microscopy Health Sciences Centre Winnipeg, Canada.
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==============================Original Headers============================== 6, 20 -- From GBurgess-at-exchange.hsc.mb.ca Fri Jan 20 16:10:02 2006 6, 20 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0KMA1mn017769 6, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Jan 2006 16:10:01 -0600 6, 20 -- Received: from hscxntmx0007.hsc.mb.ca (unverified [172.16.6.179]) by 6, 20 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 6, 20 -- {B0017584293-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Fri, 6, 20 -- 20 Jan 2006 16:09:53 -0600 6, 20 -- Received: by hscxntmx0007.hsc.mb.ca with Internet Mail Service 6, 20 -- (5.5.2653.19)id {T7CLC1WX} ; Fri, 20 Jan 2006 16:10:27 -0600 6, 20 -- Message-ID: 6, 20 -- {00A937989100304A83A058F6C45873FF32A36C-at-hscxntmx0005.hsc.mb.ca} 6, 20 -- Date: Fri, 20 Jan 2006 16:06:48 -0600 6, 20 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 6, 20 -- Subject: Wrinkled LM Sections 6, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 6, 20 -- X-Mailer: Internet Mail Service (5.5.2653.19) 6, 20 -- MIME-Version: 1.0 6, 20 -- Content-Type: text/plain; 6, 20 -- charset="iso-8859-1" ==============================End of - Headers==============================
in my experience (I am familiar with large semithin sections = up to 5 x 5 mm from human diagnostic samples for some 20 years now) your question cannot be answered with one sentence. There are several parameters to be included in a thoroughly checking of causes, starting from the type of tissue, via fixation, (full) dehydration, intermedium (full evaporation/complete exchange by resin), type, quality [polymerisation, hardness] of resin, quality of knife edge, sectioning parameters (knife, cutting angle, speed) and type of transfer of sections from the water trough to the slide, and last but not least, "special treatment" of the floating section on a water drop (i.e., for example, trying to spread sections by xylene vapor [other - more healthier - vapors?] or gentle - longer - warming up by means of a light bulb positioned above the section/slide). So IMO, you have to discriminate the problem(s) from the whole processing schedule.......(;-(.....
You tell us that you face problems not everytime, but sometimes..... Could you give us some examples (tissue type, some hints on chemicals /resin you use?) where you get such results? (not to forget the dimensions of the tissue blocks and type of knife used)....
Perhaps there is a simple solution......who knows,
best regards and have a nice weekend,
Wolfgang Muss
Salzburg, Austria
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
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I'm looking for ideas here as to what might be causing this effect. What do people think?
Garry Burgess Charge Technologist - Electron Microscopy Health Sciences Centre Winnipeg, Canada.
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==============================Original Headers============================== 6, 20 -- From GBurgess-at-exchange.hsc.mb.ca Fri Jan 20 16:10:02 2006 6, 20 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0KMA1mn017769 6, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Jan 2006 16:10:01 -0600 6, 20 -- Received: from hscxntmx0007.hsc.mb.ca (unverified [172.16.6.179]) by 6, 20 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 6, 20 -- {B0017584293-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Fri, 6, 20 -- 20 Jan 2006 16:09:53 -0600 6, 20 -- Received: by hscxntmx0007.hsc.mb.ca with Internet Mail Service 6, 20 -- (5.5.2653.19)id {T7CLC1WX} ; Fri, 20 Jan 2006 16:10:27 -0600 6, 20 -- Message-ID: 6, 20 -- {00A937989100304A83A058F6C45873FF32A36C-at-hscxntmx0005.hsc.mb.ca} 6, 20 -- Date: Fri, 20 Jan 2006 16:06:48 -0600 6, 20 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 6, 20 -- Subject: Wrinkled LM Sections 6, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 6, 20 -- X-Mailer: Internet Mail Service (5.5.2653.19) 6, 20 -- MIME-Version: 1.0 6, 20 -- Content-Type: text/plain; 6, 20 -- charset="iso-8859-1" ==============================End of - Headers==============================
==============================Original Headers============================== 25, 28 -- From W.Muss-at-salk.at Sat Jan 21 04:43:58 2006 25, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 25, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0LAhv9E003479 25, 28 -- for {Microscopy-at-microscopy.com} ; Sat, 21 Jan 2006 04:43:57 -0600 25, 28 -- Received: from localhost (localhost [127.0.0.1]) 25, 28 -- by hermes.lks.at (Postfix) with ESMTP id B185A5A9046; 25, 28 -- Sat, 21 Jan 2006 11:43:51 +0100 (CET) 25, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 25, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 25, 28 -- with ESMTP id 42900-09; Sat, 21 Jan 2006 11:43:51 +0100 (CET) 25, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 25, 28 -- by hermes.lks.at (Postfix) with SMTP id 754D65A9044; 25, 28 -- Sat, 21 Jan 2006 11:43:51 +0100 (CET) 25, 28 -- Received: by localhost with Microsoft MAPI; Sat, 21 Jan 2006 11:43:49 +0100 25, 28 -- Message-ID: {01C61E7F.F216D3E0.W.Muss-at-salk.at} 25, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 25, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 25, 28 -- To: "'GBurgess-at-exchange.hsc.mb.ca'" {GBurgess-at-exchange.hsc.mb.ca} , 25, 28 -- "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 25, 28 -- Subject: [Microscopy] Re: Wrinkled LM Sections 25, 28 -- Date: Sat, 21 Jan 2006 11:43:48 +0100 25, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 25, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 25, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 25, 28 -- MIME-Version: 1.0 25, 28 -- Content-Type: text/plain; charset="us-ascii" 25, 28 -- Content-Transfer-Encoding: 7bit 25, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
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Email: wong-at-msg.ucsf.edu Name: Mei Lie Wong
Title-Subject: [Filtered] formvar
Question: When using formvar in ethylene dichloride, is the any way to make the films slightly thicker. I know when using formvar in chloroform, you can just drain slower or faster to make films thinner or thicker. Is there something like this for the ethylene dichloride formula? I have tried several different times but so far haven't hit on anything totally satisfactory.
Mei Lie Wong wrote: ======================================================== Question: When using formvar in ethylene dichloride, is the any way to make the films slightly thicker. I know when using formvar in chloroform, you can just drain slower or faster to make films thinner or thicker. Is there something like this for the ethylene dichloride formula? I have tried several different times but so far haven't hit on anything totally satisfactory. ======================================================= This is discussed on http://www.2spi.com/catalog/submat/sup_film6.html
Disclaimer: SPI Supplies manufactures Formvar coated grids for customers and we disclose some of the tricks we use for making thicker or thinner films when using ethylene dichloride.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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==============================Original Headers============================== 10, 25 -- From cgarber-at-2spi.com Sat Jan 21 10:41:36 2006 10, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 10, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0LGfaP8025961 10, 25 -- for {microscopy-at-msa.microscopy.com} ; Sat, 21 Jan 2006 10:41:36 -0600 10, 25 -- Received: from ibm1x23g2abfyg ([71.225.72.4]) 10, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k0LGfYOZ012677; 10, 25 -- Sat, 21 Jan 2006 11:41:35 -0500 10, 25 -- X-IDV-FirstRcvd: [71.225.72.4] 10, 25 -- X-IDV-HELO: ibm1x23g2abfyg 10, 25 -- Message-ID: {01e401c61ea9$883ec840$6401a8c0-at-ibm1x23g2abfyg} 10, 25 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 10, 25 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 10, 25 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 10, 25 -- Subject: Formvar filmed grids: controlling thickness 10, 25 -- Date: Sat, 21 Jan 2006 11:41:30 -0500 10, 25 -- MIME-Version: 1.0 10, 25 -- Content-Type: text/plain; 10, 25 -- format=flowed; 10, 25 -- charset="Windows-1252"; 10, 25 -- reply-type=original 10, 25 -- Content-Transfer-Encoding: 7bit 10, 25 -- X-Priority: 3 10, 25 -- X-MSMail-Priority: Normal 10, 25 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 10, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
I was interested in the comments about banging on the sides of a sputter ion pump with a hammer or screw driver to overcome the startup problem that arises when a whisker or flake of titanium has formed and shorted out the path between the cathode and anode of the pump. Another way to treat this problem is discussed on p. 295 of my book 'Vacuum Methods in Electron Microscopy' (available from SPI, Ladd, M.E. Taylor, etc. For a description see: www.2spi.com/catalog/books/book48.html). This involves allowing the pressure in the pump to rise slightly above 1 Pa (10-2 Torr) and then turning on the high voltage power for a few seconds for three or four times. Be careful not to do this long enough to damage the power supply. -- 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============================== 1, 14 -- From bigelow-at-engin.umich.edu Sat Jan 21 11:29:43 2006 1, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 1, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0LHTgo2002820 1, 14 -- for {microscopy-at-microscopy.com} ; Sat, 21 Jan 2006 11:29:42 -0600 1, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- by srvr22.engin.umich.edu (8.13.5/8.13.5) with ESMTP id k0LHTfRg008116 1, 14 -- for {microscopy-at-microscopy.com} ; Sat, 21 Jan 2006 12:29:42 -0500 (EST) 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06210202bff81ae0e465-at-[141.212.131.221]} 1, 14 -- Date: Sat, 21 Jan 2006 12:29:41 -0500 1, 14 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 1, 14 -- Subject: [Microscopy] RE: Starting Ion Pumps 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Yes, that might also work. However, if I just got through with an 8 hour bakeout, why would I want to degrade vacuum in the gun chamber? I would either never get the chamber pumped down or it would take a very long time to reach terminal vacuum.
The idea is to get vacuum as good as possible and then isolate the gun chamber before turning on the pump. Even so, startup current would be high.
I've used the hair dryer most of the time. Then, sometimes whack the pump with the screwdriver handle. This is only for the small pumps. For some reason, they don't start as readily as the larger ones.
gary g.
At 09:48 AM 1/21/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Sat Jan 21 12:09:22 2006 10, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0LI9KuQ010814 10, 20 -- for {microscopy-at-microscopy.com} ; Sat, 21 Jan 2006 12:09:21 -0600 10, 20 -- Received: (qmail 22469 invoked from network); 21 Jan 2006 10:09:20 -0800 10, 20 -- Received: by simscan 1.1.0 ppid: 22466, pid: 22467, t: 0.1656s 10, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 20 -- by qsmtp1 with SMTP; 21 Jan 2006 10:09:20 -0800 10, 20 -- Message-Id: {6.2.3.4.2.20060121100351.020356f8-at-mail.calweb.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 10, 20 -- Date: Sat, 21 Jan 2006 10:09:22 -0800 10, 20 -- To: bigelow-at-engin.umich.edu 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] RE: Starting Ion Pumps 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200601211748.k0LHmBJV006678-at-ns.microscopy.com} 10, 20 -- References: {200601211748.k0LHmBJV006678-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Celebration of life for Delbert E. Philpott September 24, 1923 - December 11, 2005
Yesterday at the Liberty Lodge Masonic Center in Santa Clara, CA, we honored the life and friendship of Dr. Del Philpott, a lifelong microscopist and soldier who truly believed in the perpetuation of peace and worked for that goal his entire life.
His wife, Donna, agreed to our reprinting of a few of his many achievements which she listed in the memorial pamphlet.
Del was born and raised in Omro, Wisconsin. Dr. Philpott was a World War II Veteran who served with the Fighting 69th Infantry Division as they raced across Europe to link up with the Russian 58th Guards at the Elbe River in Germany in April of 1945. The famous link-up photo, by photographer Allan Jackson of 3 Americans (including Del), and 3 Russians shaking hands was printed in newspapers across the United States. The soldiers however didn't see the photo because the war hadn't yet ended. Fellow Infantryman Sam Popkins, who had helped Allan recruit soldiers for the photo, later told Del about it and confirmed that Del was one the soldiers in the photo. Del was awarded the Bronze Star and Purple Heart Medals for his service.
After the war, Del completed his Bachelor's Degree in chemistry at Indiana University in 1948 and his Master's Degree in 1949. As a Research Associate at the University of Illinois Medical School in Chicago from 1949 to 1952, he established the first electron microscope facility for the medical school. He was Head of Electron Microscopy, Marine Biological Lab and Institute for Muscle Research, Woods Hole, MA from 1952 to 1963, where he established the electron microscope facility for the 2 Institutes.
A pioneer in the field of Electron Microscopy, he directed and ran research projects under Nobel Laureate Dr. Albert Szent-Gyorgyi in the winter and for the Marine Biological Lab at Woods Hole during the summer. He built his own ultramicrotome for ultra-thin sectioning.
He published the first paper on ultrastructure of the human heart with Dr. Bruno Kish and published the first electron micrographs of a protein crystal isolated from muscle. He demonstrated that ribosomes travel from the nucleus to the cytoplasm via the nuclear pores. Del got his PhD in cytology in 1963 at Boston University.
While Professor of Biochemistry at the University of Colorado from 1963 to 1965, he established and ran the electron microscope laboratory at the medical school. In 1966, he was Head of the Department of Electron Microscopy and Co-Director of the Institute for Biomedical Research at Mercy Hospital in Denver, CO.
Del came to California in the 60's to work as a Research Scientist as the Head of the Ultrastructure Laboratory at the Moffett Field NASA Ames Research Center. He inspected lunar soil for signs of life and had experiments flown on both American and Russian spacecraft including Apollo 17, Cosmos 736, Cosmos 936, and SL-3. He developed a fixative for space flight that preserves the ultrastructure of tissues for over 4 years without the need for embedding. Del retired from NASA in 1990, but worked under contract with the Lockheed Martin Company at NASA Ames as a Research Associate whose duties included assisting visiting Russian scientists with space related experiments.
During his professional career, Del authored over 230 scientific papers and wrote articles for several books and magazines.
In 1995, Del and his wife Donna co-edited the book "Hands Across The Elbe", stories by American and Russian veterans about the link up that cut Fascist Germany in half in April 1945.
Del was one of 10 veterans chosen to be guests at the Russian government's May 9, 2005, 60th Anniversary Celebration of the end of World War II. He was selected to be seated at the table with the President of China, President Putin, and President and Mrs. Bush at the Kremlin Reception following the Parade e in Red Square.
Del's interests included flying, raising dogs, ham radio, ceramics, spinning, photography, woodcarving, magic, and travel. He held offices in many organizations including several scientific societies, veterans' organizations, and The Unique Boutique at the Sunnyvale Senior Center. He was a member of Liberty Lodge $299, Valley Star Chapter #250 O.E.S., and the San Jose Scottish Rite.
He wanted to be remembered for his part in developing interaction between the American and Russian scientists in space research. He believed that interactions among the citizens of all nations are necessary for the perpetuation of peace and felt that the "Spirit of the Elbe" encourages this vision.
Thank you Donna for putting together such an information-packed bibliography.
I knew Del from the late 60's from the microscopy society. He was a wonderful friend and had an uncanny sense of humor. I would like to share 2 of Del's favorite stories. And for those of you who knew Del, you know he had LOTS of stories.
I can no longer remember the exact year, but it was in the 70's. Someone sent in a bogus abstract for the EMSA meeting with a crazy title and crazy names of authors. Unfortunately I no longer remember those either. However the following year, as you can imagine, EMSA was trying to review the abstracts or at least look more carefully at the titles and authors. That year Del sent in an abstract about research on lunar samples. Well, whoever reviewed his abstract decided that no one could possibly be named Delbert Philpott and certainly no one was looking at moon samples, SO, his paper was rejected!!
Del's sense of humor was ever present. Before he knew Donna, again in the 70's sometime, Del was at the EMSA meeting telling us about a Valentine's present he got for his girlfriend. He bought some dog biscuits (yes dog biscuits), had them chocolate coated, put them in those fancy white wrappers, and put them in a standard Valentine's Candy Box. He gave them to his girlfriend. The next year I saw Del, I asked him how his girlfriend liked her Valentine's gift. He said "Well, I actually haven't seen her since then. I guess she didn't like that brand dog biscuits"!!!!
Del is gone, but his loving sense of humor, and desire for the promotion of peace through interactions among citizens of all nations will always be with us. We love you Del, and will miss you always.
God Bless Us All, Judy Murphy
==============================Original Headers============================== 21, 17 -- From murphyjudy-at-comcast.net Sun Jan 22 17:23:21 2006 21, 17 -- Received: from sccrmhc11.comcast.net (sccrmhc11.comcast.net [63.240.77.81]) 21, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0MNNK1W018752 21, 17 -- for {Microscopy-at-microscopy.com} ; Sun, 22 Jan 2006 17:23:21 -0600 21, 17 -- Received: from [192.168.1.102] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 21, 17 -- by comcast.net (sccrmhc11) with ESMTP 21, 17 -- id {2006012223231901100qjb1ie} ; Sun, 22 Jan 2006 23:23:19 +0000 21, 17 -- Message-ID: {43D413E6.6010102-at-comcast.net} 21, 17 -- Date: Sun, 22 Jan 2006 15:23:18 -0800 21, 17 -- From: Judy Murphy {murphyjudy-at-comcast.net} 21, 17 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 21, 17 -- X-Accept-Language: en-us, en 21, 17 -- MIME-Version: 1.0 21, 17 -- To: Microscopy {Microscopy-at-microscopy.com} 21, 17 -- Subject: We Remember Del Philpott 21, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 21, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Dear Gary, I have been working with LR white and have been facing similar problems. They are more so only when I immuno label those sections. Then again my sections are more than 2 mm in size and they are thin sections. I have managed to reduce the wrinkles by changing the washing techniques between immunolabelling steps but that hasnt completely eliminated the wrinkles. I have observed that LR white section around 1mm wide didnt have so many wrinkles. Unfortunately for me I cant have thicker sections or smaller sections to avoid this problem. I cannot use a different resin since it interfers with my immunolabeling. SO the bottom line being, I have learnt to live with it unfortunately. But I have a feeling that smaller sections do better and the wrinsing the grids in a drop of water/ buffer on a petridish does help reduce the wrinkles greatly if not eliminate it.
Any advice on photoshop micracles of ironing out the wrinkles would be appreciated. I hope facelifting TEM pics will not be frowned upon by the microscopy community.
Looking forward for comments and suggestion.
Regards, Vinod Nair Graduate Student Dept. of Biology New Mexico State University
On 1/21/06, W.Muss-at-salk.at {W.Muss-at-salk.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 } ---------------------------------------------------------------------------- } } Good morning, all, } } Dear Garry, } } in my experience (I am familiar with large semithin sections = up to 5 x 5 } mm from human diagnostic samples for some 20 years now) your question } cannot be answered with one sentence. } There are several parameters to be included in a thoroughly checking of } causes, starting from the type of tissue, via fixation, (full) dehydration, } intermedium (full evaporation/complete exchange by resin), type, quality } [polymerisation, hardness] of resin, quality of knife edge, sectioning } parameters (knife, cutting angle, speed) and type of transfer of sections } from the water trough to the slide, and last but not least, "special } treatment" of the floating section on a water drop (i.e., for example, } trying to spread sections by xylene vapor [other - more healthier - } vapors?] or gentle - longer - warming up by means of a light bulb } positioned above the section/slide). } So IMO, you have to discriminate the problem(s) from the whole processing } schedule.......(;-(..... } } You tell us that you face problems not everytime, but sometimes..... } Could you give us some examples (tissue type, some hints on chemicals } /resin you use?) where you get such results? (not to forget the dimensions } of the tissue blocks and type of knife used).... } } Perhaps there is a simple solution......who knows, } } } best regards and have a nice weekend, } } Wolfgang Muss } } Salzburg, Austria } } Paracelsus Medical Private University (PMU) } Institute of Pathology } Electron Microscopy Lab } Muellner Hauptstrasse 48 } A-5020 SALZBURG, Austria/Europe } Phone work: +43+662+4482+4720 } Mobile phone work:+43+662+4482-57704 } Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o } W.Muss") } E-Mail work: W.Muss-at-SALK.at } } Mobile-phone private: ++43+676+5 369-456 } E-Mail private: wij.Muss-at-aon.at } ------------------------------------------------------------------------ } --------------------------------Information on behalf of } Society for Cutaneous Ultrastructure Research (SCUR) } PLEASE VISIT THE UPDATED WEBSITE of SCUR at } } http://www.scur.org { } ------------------------------------------------------------------------- } Forthcoming Meetings: } 33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland } WEBSITE, containing all FORMS: http://www.scur.org.pl } Additional informations: send an E-Mail } kwoznia-at-amwaw.edu.pl } ---------------------- } 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE } Czech Republic } ---------------------- } 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan } Joint Meeting with the } JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology } } } } } } } ---------- } Von: GBurgess-at-exchange.hsc.mb.ca[SMTP:GBurgess-at-exchange.hsc.mb.ca] } Antwort an: GBurgess-at-exchange.hsc.mb.ca } Gesendet: Freitag, 20. Janner 2006 23:52 } An: W.Muss-at-salk.at } Betreff: [Microscopy] Wrinkled LM Sections } } ------------------------------------------------------------------------ } ---- } 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 } ------------------------------------------------------------------------ } ---- } Even though I am very experienced in Electron Microscopy, there are times } when I cut LM sections that are extremely wrinkled, and they look like } cracked glass. I normally don't experience this, but when it happens, even } considering every possible variable, I'm still not exactly sure what might } be at work here to give this sort of result. } } I'm looking for ideas here as to what might be causing this effect. What } do } people think? } } } Garry Burgess } Charge Technologist - Electron Microscopy } Health Sciences Centre } Winnipeg, Canada. } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. } } ==============================Original } Headers============================== } 6, 20 -- From GBurgess-at-exchange.hsc.mb.ca Fri Jan 20 16:10:02 2006 } 6, 20 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca } [142.233.100.122]) } 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k0KMA1mn017769 } 6, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Jan 2006 16:10:01 -0600 } 6, 20 -- Received: from hscxntmx0007.hsc.mb.ca (unverified [172.16.6.179]) } by } 6, 20 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id } 6, 20 -- {B0017584293-at-hscxntmx0003.hsc.mb.ca} for } {Microscopy-at-microscopy.com} ;Fri, } 6, 20 -- 20 Jan 2006 16:09:53 -0600 } 6, 20 -- Received: by hscxntmx0007.hsc.mb.ca with Internet Mail Service } 6, 20 -- (5.5.2653.19)id {T7CLC1WX} ; Fri, 20 Jan 2006 16:10:27 -0600 } 6, 20 -- Message-ID: } 6, 20 -- {00A937989100304A83A058F6C45873FF32A36C-at-hscxntmx0005.hsc.mb.ca} } 6, 20 -- Date: Fri, 20 Jan 2006 16:06:48 -0600 } 6, 20 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} } 6, 20 -- Subject: Wrinkled LM Sections } 6, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} } 6, 20 -- X-Mailer: Internet Mail Service (5.5.2653.19) } 6, 20 -- MIME-Version: 1.0 } 6, 20 -- Content-Type: text/plain; } 6, 20 -- charset="iso-8859-1" } ==============================End of - } Headers============================== } } } } ==============================Original Headers============================== } 25, 28 -- From W.Muss-at-salk.at Sat Jan 21 04:43:58 2006 } 25, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) } 25, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0LAhv9E003479 } 25, 28 -- for {Microscopy-at-microscopy.com} ; Sat, 21 Jan 2006 04:43:57 -0600 } 25, 28 -- Received: from localhost (localhost [127.0.0.1]) } 25, 28 -- by hermes.lks.at (Postfix) with ESMTP id B185A5A9046; } 25, 28 -- Sat, 21 Jan 2006 11:43:51 +0100 (CET) } 25, 28 -- Received: from hermes.lks.at ([127.0.0.1]) } 25, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) } 25, 28 -- with ESMTP id 42900-09; Sat, 21 Jan 2006 11:43:51 +0100 (CET) } 25, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) } 25, 28 -- by hermes.lks.at (Postfix) with SMTP id 754D65A9044; } 25, 28 -- Sat, 21 Jan 2006 11:43:51 +0100 (CET) } 25, 28 -- Received: by localhost with Microsoft MAPI; Sat, 21 Jan 2006 11:43:49 +0100 } 25, 28 -- Message-ID: {01C61E7F.F216D3E0.W.Muss-at-salk.at} } 25, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} } 25, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} } 25, 28 -- To: "'GBurgess-at-exchange.hsc.mb.ca'" {GBurgess-at-exchange.hsc.mb.ca} , } 25, 28 -- "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} } 25, 28 -- Subject: [Microscopy] Re: Wrinkled LM Sections } 25, 28 -- Date: Sat, 21 Jan 2006 11:43:48 +0100 } 25, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} } 25, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor } 25, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 } 25, 28 -- MIME-Version: 1.0 } 25, 28 -- Content-Type: text/plain; charset="us-ascii" } 25, 28 -- Content-Transfer-Encoding: 7bit } 25, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at } ==============================End of - Headers============================== }
==============================Original Headers============================== 6, 25 -- From nairvinods-at-gmail.com Sun Jan 22 17:45:05 2006 6, 25 -- Received: from zproxy.gmail.com (zproxy.gmail.com [64.233.162.195]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0MNj4D3022636 6, 25 -- for {microscopy-at-microscopy.com} ; Sun, 22 Jan 2006 17:45:04 -0600 6, 25 -- Received: by zproxy.gmail.com with SMTP id 9so805621nzo 6, 25 -- for {microscopy-at-microscopy.com} ; Sun, 22 Jan 2006 15:45:03 -0800 (PST) 6, 25 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 25 -- s=beta; d=gmail.com; 6, 25 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 6, 25 -- b=iwLI/yOhyn8RBFdLsd4GrjjADMj8yG6x8a+cZRddAPXP313KWFTH8mhP3WJuqdwmBf56pLTJ6QHi6KG7BMIsRtxOEYY4fvwY5oFvcx+g2Qe17GhH6Q6gSFUGuaU44HZYNriuBxtZcmHsPNQ9H4Cu/ddrt5xiiMyNw1HrJ33u1+I= 6, 25 -- Received: by 10.64.251.4 with SMTP id y4mr2436900qbh; 6, 25 -- Sun, 22 Jan 2006 15:38:52 -0800 (PST) 6, 25 -- Received: by 10.64.201.14 with HTTP; Sun, 22 Jan 2006 15:38:52 -0800 (PST) 6, 25 -- Message-ID: {ea42a3900601221538n63c2c777pab547602772aa678-at-mail.gmail.com} 6, 25 -- Date: Sun, 22 Jan 2006 16:38:52 -0700 6, 25 -- From: Vinod Nair {nairvinods-at-gmail.com} 6, 25 -- To: microscopy-at-microscopy.com 6, 25 -- Subject: Re: [Microscopy] Re: Wrinkled LM Sections 6, 25 -- In-Reply-To: {200601211126.k0LBQBcT011510-at-ns.microscopy.com} 6, 25 -- MIME-Version: 1.0 6, 25 -- Content-Type: text/plain; charset=UTF-8 6, 25 -- Content-Disposition: inline 6, 25 -- References: {200601211126.k0LBQBcT011510-at-ns.microscopy.com} 6, 25 -- Content-Transfer-Encoding: 8bit 6, 25 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k0MNj4D3022636 ==============================End of - Headers==============================
Mei Lie, Making formvar films in ethylene dichloride We used these films for 30 yrs and controlled the thickness mostly by the percentage of formvar to ethylene dichloride. What percentage are you using?
Judy
Judy Murphy, PhD Microscopy Training, Imaging, and Lab Design Stockton, CA 95219 murphyjudy-at-comcast.net
wong-at-msg.ucsf.edu wrote:
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==============================Original Headers============================== 12, 19 -- From murphyjudy-at-comcast.net Sun Jan 22 22:53:43 2006 12, 19 -- Received: from sccrmhc12.comcast.net (sccrmhc12.comcast.net [63.240.77.82]) 12, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0N4rhWv009022 12, 19 -- for {microscopy-at-microscopy.com} ; Sun, 22 Jan 2006 22:53:43 -0600 12, 19 -- Received: from [192.168.1.102] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 12, 19 -- by comcast.net (sccrmhc12) with ESMTP 12, 19 -- id {20060123045340012005vqqbe} ; Mon, 23 Jan 2006 04:53:41 +0000 12, 19 -- Message-ID: {43D46116.2070908-at-comcast.net} 12, 19 -- Date: Sun, 22 Jan 2006 20:52:38 -0800 12, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 12, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 12, 19 -- X-Accept-Language: en-us, en 12, 19 -- MIME-Version: 1.0 12, 19 -- To: Microscopy List Server {microscopy-at-microscopy.com} 12, 19 -- Subject: Re: [Microscopy] viaWWW: Making formvar thicker 12, 19 -- References: {200601211425.k0LEP0O9019151-at-ns.microscopy.com} 12, 19 -- In-Reply-To: {200601211425.k0LEP0O9019151-at-ns.microscopy.com} 12, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
A web site was brought to my attention that is a collection of photographs by him (and several of him) during Del's time with Albert Szent-Gyorgyi. I found it very interesting and rich in the history of science.
Thank you Peter for sharing the information AND hold on to that signed copy of Crossing the Elbe!
murphyjudy-at-comcast.net wrote:
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==============================Original Headers============================== 8, 19 -- From murphyjudy-at-comcast.net Sun Jan 22 23:08:21 2006 8, 19 -- Received: from sccrmhc14.comcast.net (sccrmhc14.comcast.net [63.240.77.84]) 8, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0N58KXw011747 8, 19 -- for {microscopy-at-microscopy.com} ; Sun, 22 Jan 2006 23:08:20 -0600 8, 19 -- Received: from [192.168.1.102] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 8, 19 -- by comcast.net (sccrmhc14) with ESMTP 8, 19 -- id {2006012305081301400kjrige} ; Mon, 23 Jan 2006 05:08:14 +0000 8, 19 -- Message-ID: {43D464BD.4010206-at-comcast.net} 8, 19 -- Date: Sun, 22 Jan 2006 21:08:13 -0800 8, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 8, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 8, 19 -- X-Accept-Language: en-us, en 8, 19 -- MIME-Version: 1.0 8, 19 -- To: Microscopy List Server {microscopy-at-microscopy.com} 8, 19 -- Subject: Re: [Microscopy] We Remember Del Philpott 8, 19 -- References: {200601222350.k0MNoOo1024081-at-ns.microscopy.com} 8, 19 -- In-Reply-To: {200601222350.k0MNoOo1024081-at-ns.microscopy.com} 8, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hi: I am restoring a Zeiss 109 TEM where the Leybold IZ-80 iongetter pump has been removed, however they left the magnets. The pump does not have to be in working condition. If you have on lying around and are willing to part with it or sell it please let me know. Peter Jordan/EMSI
==============================Original Headers============================== 2, 22 -- From pjordan-at-dslextreme.com Sun Jan 22 23:53:27 2006 2, 22 -- Received: from mail5.dslextreme.com (mail5.dslextreme.com [66.51.199.81]) 2, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0N5rQiS023856 2, 22 -- for {microscopy-at-msa.microscopy.com} ; Sun, 22 Jan 2006 23:53:26 -0600 2, 22 -- Received: (qmail 30488 invoked from network); 23 Jan 2006 05:52:12 -0000 2, 22 -- Received: from unknown (HELO DCPZ5N31) (68.183.94.207) 2, 22 -- by mail5.dslextreme.com with SMTP; Sun, 22 Jan 2006 21:52:12 -0800 2, 22 -- Message-ID: {005401c61fe1$5218af90$cf5eb744-at-DCPZ5N31} 2, 22 -- From: "Peter Jordan" {pjordan-at-dslextreme.com} 2, 22 -- To: "Electron Microscopy" {Microscopy-at-msa.microscopy.com} 2, 22 -- Subject: Iongetter pump for Zeiss 109 2, 22 -- Date: Sun, 22 Jan 2006 21:53:22 -0800 2, 22 -- MIME-Version: 1.0 2, 22 -- Content-Type: text/plain; 2, 22 -- format=flowed; 2, 22 -- charset="iso-8859-1"; 2, 22 -- reply-type=original 2, 22 -- Content-Transfer-Encoding: 7bit 2, 22 -- X-Priority: 3 2, 22 -- X-MSMail-Priority: Normal 2, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 2, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
Dear Gary, Let me begin with apologising for not reading the email correctly. You can see what I have been pondering over from my reply with regards to LR white resin:( Guess it was desperation that made me misread LM as LR and link it to the wrinkeling problems I have been facing. Having said that I feel very foolish now for being hasty and replying to your email query. My sincere apologies.
But then again I am open for suggestions,comments or insight into the problem I am facing. regards, Vinod
==============================Original Headers============================== 3, 25 -- From nairvinods-at-gmail.com Mon Jan 23 00:03:21 2006 3, 25 -- Received: from wproxy.gmail.com (wproxy.gmail.com [64.233.184.196]) 3, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0N63JfS026967 3, 25 -- for {microscopy-at-microscopy.com} ; Mon, 23 Jan 2006 00:03:20 -0600 3, 25 -- Received: by wproxy.gmail.com with SMTP id i30so857046wra 3, 25 -- for {microscopy-at-microscopy.com} ; Sun, 22 Jan 2006 22:03:19 -0800 (PST) 3, 25 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 3, 25 -- s=beta; d=gmail.com; 3, 25 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 3, 25 -- b=qLDCg7ZEROXSfD8aXh3sQ7gV08fkTVY98jzE998c6AOreIritxcbW8wfosf9yDu9vRIfZK7mOZoljeb0to6Wf+TmtvFGfObfREqnJAWvRbc9PTSIm6Ow+fqo0HEEIcqZzzbUyMrn61hqiKideomsMmSKQsDqs0bm8+Hne14//ZM= 3, 25 -- Received: by 10.64.143.19 with SMTP id q19mr271269qbd; 3, 25 -- Sun, 22 Jan 2006 21:57:15 -0800 (PST) 3, 25 -- Received: by 10.64.201.14 with HTTP; Sun, 22 Jan 2006 21:57:15 -0800 (PST) 3, 25 -- Message-ID: {ea42a3900601222157v6fe56138p3a4d08b1e4b0777a-at-mail.gmail.com} 3, 25 -- Date: Sun, 22 Jan 2006 22:57:15 -0700 3, 25 -- From: Vinod Nair {nairvinods-at-gmail.com} 3, 25 -- To: microscopy-at-microscopy.com 3, 25 -- Subject: Re: [Microscopy] Wrinkled LM Sections 3, 25 -- In-Reply-To: {200601230040.k0N0elR9004151-at-ns.microscopy.com} 3, 25 -- MIME-Version: 1.0 3, 25 -- Content-Type: text/plain; charset=UTF-8 3, 25 -- Content-Disposition: inline 3, 25 -- References: {200601230040.k0N0elR9004151-at-ns.microscopy.com} 3, 25 -- Content-Transfer-Encoding: 8bit 3, 25 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k0N63JfS026967 ==============================End of - Headers==============================
On Jan 21, 2006, at 6:03 AM, wong-at-msg.ucsf.edu wrote:
} Question: When using formvar in ethylene dichloride, is the any way to } make the films slightly thicker. I know when using formvar in } chloroform, you can just drain slower or faster to make films thinner } or thicker. Is there something like this for the ethylene dichloride } formula? I have tried several different times but so far haven't hit } on anything totally satisfactory. } Dear Mei Lie, Using a more concentrated formvar solution should do it, and if you only have the pre-disolved formvar, you could try either letting some of the solvent evaporate or double dipping. For finer variations you might try letting the formvar drain at a shallower angle, which will drain it slightly slower. Good luck. 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 Mon Jan 23 11:37:40 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0NHbemg016835 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 23 Jan 2006 11:37:40 -0600 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 7071C109F3B 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 23 Jan 2006 09:37:39 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id E6173109E19 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 23 Jan 2006 09:37:37 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200601211403.k0LE3NrZ014761-at-ns.microscopy.com} 4, 22 -- References: {200601211403.k0LE3NrZ014761-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {6675a89478c5c6f5c36fc2d5e8dcd7a0-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: Making formvar thicker 4, 22 -- Date: Mon, 23 Jan 2006 09:44:45 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Dear All, We have an older Zeiss Opni-1 surgical microscope and we'd like to replace its broken tungsten illuminator with a fiber optic illuminator. this would preserve the large illuminated field diameter and be adjustable in angle. the usual web searches have shown that retrofits were made for this scope, but haven't provided any useful information on performance or options. It is already used in conjunction with bifurcated light guides, but we have users who need the stock style of illumination.
Thanks, Glen
Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
==============================Original Headers============================== 7, 23 -- From glenmac-at-u.washington.edu Mon Jan 23 11:39:38 2006 7, 23 -- Received: from mxout2.cac.washington.edu (mxout2.cac.washington.edu [140.142.33.4]) 7, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0NHdbYl017204 7, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 11:39:37 -0600 7, 23 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 7, 23 -- by mxout2.cac.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k0NHdaNw018071 7, 23 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 7, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 09:39:37 -0800 7, 23 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 7, 23 -- (authenticated authid=glenmac) 7, 23 -- by smtp.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k0NHdac2004853 7, 23 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 7, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 09:39:36 -0800 7, 23 -- Mime-Version: 1.0 (Apple Message framework v746.2) 7, 23 -- Content-Transfer-Encoding: 7bit 7, 23 -- Message-Id: {E3E326B4-892B-4DAE-BA4A-8D7CA81A632C-at-u.washington.edu} 7, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 23 -- To: MSA {Microscopy-at-MSA.Microscopy.Com} 7, 23 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 7, 23 -- Subject: fiber illuminator for Opni-1 7, 23 -- Date: Mon, 23 Jan 2006 09:39:32 -0800 7, 23 -- X-Mailer: Apple Mail (2.746.2) 7, 23 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 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' ==============================End of - Headers==============================
I need to measure thermal conductivity of thin, 50-200 nm film. I would appreciate any suggestions regarding to equipment, techniques, and literature references.
Thank you,
Victoria Fink vfink-at-shaw.ca
==============================Original Headers============================== 7, 28 -- From vfink-at-shaw.ca Mon Jan 23 12:04:15 2006 7, 28 -- Received: from pd5mo1so.prod.shaw.ca (shawidc-mo1.cg.shawcable.net [24.71.223.10]) 7, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0NI4FmR023679 7, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 12:04:15 -0600 7, 28 -- Received: from pd2mr7so.prod.shaw.ca (pd2mr7so-qfe3.prod.shaw.ca [10.0.141.10]) 7, 28 -- by l-daemon (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 15 2004)) 7, 28 -- with ESMTP id {0ITK00H9N4UXI290-at-l-daemon} for Microscopy-at-MSA.Microscopy.Com; 7, 28 -- Mon, 23 Jan 2006 11:04:09 -0700 (MST) 7, 28 -- Received: from pn2ml1so.prod.shaw.ca ([10.0.121.145]) 7, 28 -- by pd2mr7so.prod.shaw.ca (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 7, 28 -- 15 2004)) with ESMTP id {0ITK00JKF4UX7300-at-pd2mr7so.prod.shaw.ca} for 7, 28 -- Microscopy-at-MSA.Microscopy.Com; Mon, 23 Jan 2006 11:04:09 -0700 (MST) 7, 28 -- Received: from homep5whcwsdwx ([70.68.254.15]) 7, 28 -- by l-daemon (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 15 2004)) 7, 28 -- with SMTP id {0ITK001EW4UVGR10-at-l-daemon} for Microscopy-at-MSA.Microscopy.Com; 7, 28 -- Mon, 23 Jan 2006 11:04:09 -0700 (MST) 7, 28 -- Date: Mon, 23 Jan 2006 10:04:11 -0800 7, 28 -- From: Victoria Fink {vfink-at-shaw.ca} 7, 28 -- Subject: thermal conductivity of thin film measurement techniques 7, 28 -- To: MSA listserver {Microscopy-at-MSA.Microscopy.Com} 7, 28 -- Message-id: {042001c62047$69d10270$0ffe4446-at-homep5whcwsdwx} 7, 28 -- MIME-version: 1.0 7, 28 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 7, 28 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 7, 28 -- Content-type: text/plain; format=flowed; charset=iso-8859-1; reply-type=response 7, 28 -- Content-transfer-encoding: 7bit 7, 28 -- X-Priority: 3 7, 28 -- X-MSMail-priority: Normal ==============================End of - Headers==============================
Inked to measure thermal conductivity of thin, 50-200 nm film. I would appreciate any suggestions regarding to equipment, techniques, and literature references. Thank you,
Victoria Fink vfink-at-shaw.ca
==============================Original Headers============================== 6, 29 -- From vfink-at-shaw.ca Mon Jan 23 12:04:23 2006 6, 29 -- Received: from pd4mo1so.prod.shaw.ca (shawidc-mo1.cg.shawcable.net [24.71.223.10]) 6, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0NI4L6j023721 6, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 12:04:22 -0600 6, 29 -- Received: from pd3mr2so.prod.shaw.ca 6, 29 -- (pd3mr2so-qfe3.prod.shaw.ca [10.0.141.178]) by l-daemon 6, 29 -- (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 15 2004)) 6, 29 -- with ESMTP id {0ITK00MFH4TCBF80-at-l-daemon} for Microscopy-at-MSA.Microscopy.Com; 6, 29 -- Mon, 23 Jan 2006 11:03:12 -0700 (MST) 6, 29 -- Received: from pn2ml1so.prod.shaw.ca ([10.0.121.145]) 6, 29 -- by pd3mr2so.prod.shaw.ca (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 6, 29 -- 15 2004)) with ESMTP id {0ITK00BYS4TCXT10-at-pd3mr2so.prod.shaw.ca} for 6, 29 -- Microscopy-at-MSA.Microscopy.Com; Mon, 23 Jan 2006 11:03:12 -0700 (MST) 6, 29 -- Received: from homep5whcwsdwx ([70.68.254.15]) 6, 29 -- by l-daemon (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 15 2004)) 6, 29 -- with SMTP id {0ITK00FRN4TAXA50-at-l-daemon} for Microscopy-at-MSA.Microscopy.Com; 6, 29 -- Mon, 23 Jan 2006 11:03:12 -0700 (MST) 6, 29 -- Date: Mon, 23 Jan 2006 10:03:14 -0800 6, 29 -- From: Victoria Fink {vfink-at-shaw.ca} 6, 29 -- Subject: thermal conductivity of thin film measurement techniques 6, 29 -- To: MSA listserver {Microscopy-at-MSA.Microscopy.Com} 6, 29 -- Message-id: {041a01c62047$47e848d0$0ffe4446-at-homep5whcwsdwx} 6, 29 -- MIME-version: 1.0 6, 29 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 6, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 6, 29 -- Content-type: text/plain; format=flowed; charset=iso-8859-1; reply-type=original 6, 29 -- Content-transfer-encoding: 7bit 6, 29 -- X-Priority: 3 6, 29 -- X-MSMail-priority: Normal ==============================End of - Headers==============================
A number of our customers for coated grids request a variety of thicknesses. What we do for thicker coated grids is increase the percentage of formvar in ethylene dichloride.
Mike Bouchard
Disclaimer: Ladd Research sells Formvar coated grids and the essentials to make your own.
At 09:46 AM 1/21/2006, wong-at-msg.ucsf.edu wrote:
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I am curious about the possible skinkage artifact that may or may not occur when cryoprotecting lightly fixed (4% para / 0.1% glut) watery tissues (embryonic skin) in 20% PVP / 1.7M sucrose for ultrathin cryomicrotomy? Does the cryoprotectant replace the water in a controlled way and keep the tissue fat and happy or does it draw the water out and collapse the tissue? I would love any information on this subject or a possible reference.
Robert Underwood University of Washington Dermatology
==============================Original Headers============================== 4, 20 -- From underwoo-at-u.washington.edu Mon Jan 23 13:08:13 2006 4, 20 -- Received: from mxout7.cac.washington.edu (mxout7.cac.washington.edu [140.142.32.178]) 4, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0NJ8Anx013006 4, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 13:08:11 -0600 4, 20 -- Received: from hymn02.u.washington.edu (hymn02.u.washington.edu [140.142.13.239]) 4, 20 -- by mxout7.cac.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k0NJ89DN018919 4, 20 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 4, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 11:08:09 -0800 4, 20 -- Received: from localhost (localhost [127.0.0.1]) 4, 20 -- by hymn02.u.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k0NJ894u023894 4, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jan 2006 11:08:09 -0800 4, 20 -- X-Auth-Received: from [128.208.106.163] by hymn02.u.washington.edu via HTTP; Mon, 23 Jan 2006 11:08:09 PST 4, 20 -- Date: Mon, 23 Jan 2006 11:08:09 -0800 (PST) 4, 20 -- From: Robert A Underwood {underwoo-at-u.washington.edu} 4, 20 -- To: Microscopy List {Microscopy-at-MSA.Microscopy.Com} 4, 20 -- Subject: [Microscopy] Cryoprotection causing shrinkage artifact 4, 20 -- Message-ID: {Pine.LNX.4.43.0601231108090.3305-at-hymn02.u.washington.edu} 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 20 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
Postdoctoral Position, Department of Materials Science and Engineering Lehigh University,
Available March 2006. Opening for post-doc to work on project involving processing of patterned sapphire substrates via oxidation and solid state conversion of a metallic Al coating. Person would be responsible for processing and EBSD/TEM characterization of both the substrates, together with TEM of MOCVD grown GaN layers to determine the defect density. Expertise in materials processing and electron microscopy required, experience with thin film processing a plus. Please forward resume by e-mail to Prof. Helen Chan (Helen.Chan-at-lehigh.edu) and / or Prof. Richard Vinci (rpv2-at-lehigh.edu).
...........
Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195
==============================Original Headers============================== 5, 22 -- From jae5-at-lehigh.edu Mon Jan 23 13:25:44 2006 5, 22 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0NJPhoS019690 5, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 23 Jan 2006 13:25:44 -0600 5, 22 -- Received: from [128.180.55.141] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 5, 22 -- (authenticated bits=0) 5, 22 -- by rain.CC.Lehigh.EDU (8.13.5/8.13.5) with ESMTP id k0NJPhEc031735 5, 22 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 5, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 23 Jan 2006 14:25:43 -0500 5, 22 -- Message-ID: {43D52DB7.9050707-at-lehigh.edu} 5, 22 -- Date: Mon, 23 Jan 2006 14:25:43 -0500 5, 22 -- From: Alwyn Eades {jae5-at-lehigh.edu} 5, 22 -- Organization: Lehigh University 5, 22 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 5, 22 -- X-Accept-Language: en-us, en 5, 22 -- MIME-Version: 1.0 5, 22 -- To: "MicroscopyListserver (E-mail)" {Microscopy-at-microscopy.com} 5, 22 -- Subject: Post doctoral position at Lehigh 5, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Virus-Scanned: ClamAV version 0.88, clamav-milter version 0.87 on rain.CC.Lehigh.EDU 5, 22 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I have a colleague who recently experienced some down time on his SEM. The system went through repeated cycles of pump down, and venting and was eventually left powered down while waiting on a computer replacement.
After getting everything operating again, an oil film was found that had built up on the x-ray detector thin window. This was initially causing an unacceptable amount of background in the 0-3kV range of the x-ray signal as well as an overall low count rate. The collimator was removed and cleaned, which corrected the noise and count rate. They are in the process of replacing the roughing lines and cleaning the chamber to prevent further contamination. Now, when calibrating with a copper standard, the ratio of the L line signal versus the K lines is dramatically favoring the L. This was not the case before.
Any ideas why this might be happening and how to correct it?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 5, 23 -- From hagglundk1-at-nku.edu Tue Jan 24 09:02:19 2006 5, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0OF2JlW001861 5, 23 -- for {microscopy-at-microscopy.com} ; Tue, 24 Jan 2006 09:02:19 -0600 5, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 5, 23 -- Tue, 24 Jan 2006 10:02:19 -0500 5, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 5, 23 -- Content-class: urn:content-classes:message 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; 5, 23 -- charset="us-ascii" 5, 23 -- Subject: SEM x-ray peak ratios 5, 23 -- Date: Tue, 24 Jan 2006 10:02:18 -0500 5, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3585FA-at-mailfac1.hh.nku.edu} 5, 23 -- X-MS-Has-Attach: 5, 23 -- X-MS-TNEF-Correlator: 5, 23 -- Thread-Topic: SEM x-ray peak ratios 5, 23 -- Thread-Index: AcYg9yuMm942rGb1QfKCevfjxyMD/g== 5, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- X-OriginalArrivalTime: 24 Jan 2006 15:02:19.0222 (UTC) FILETIME=[2BE72F60:01C620F7] 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0OF2JlW001861 ==============================End of - Headers==============================
Does anyone remember a quick and easy way to calibrate - it has what 25 taps - so if anyone remembers how to do the calculations, please let me know ASAP. Thanks Barbara
==============================Original Headers============================== 1, 20 -- From maloneyb-at-fiu.edu Tue Jan 24 13:40:30 2006 1, 20 -- Received: from imappopsmtp.fiu.edu (imappopsmtp.fiu.edu [131.94.74.203]) 1, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0OJeTel015388 1, 20 -- for {microscopy-at-sparc5.microscopy.com} ; Tue, 24 Jan 2006 13:40:30 -0600 1, 20 -- Received: from [131.94.95.156] by imappopsmtp.fiu.edu 1, 20 -- (InterMail vK.4.04.00.00 201-232-137 license dcade68804da15b88628cbdb48d26a54) 1, 20 -- with SMTP 1, 20 -- id {20060124194058.VXOU22222.imappopsmtp-at-[131.94.95.156]} 1, 20 -- for {microscopy-at-sparc5.microscopy.com} ; 1, 20 -- Tue, 24 Jan 2006 14:40:58 -0500 1, 20 -- Message-ID: {43D68175.1080106-at-fiu.edu} 1, 20 -- Date: Tue, 24 Jan 2006 14:35:17 -0500 1, 20 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 20 -- X-Accept-Language: en-us, en 1, 20 -- MIME-Version: 1.0 1, 20 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-ns.microscopy.com} 1, 20 -- Subject: Phillips 300 TEM calibration 1, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I have had a request from a user looking for a cryo- ultramicrotomy workshop. Is anyone offering one in the next year or know of one? Thanks!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
==============================Original Headers============================== 5, 23 -- From edelmare-at-muohio.edu Tue Jan 24 14:11:13 2006 5, 23 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0OKBBkX020625 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 24 Jan 2006 14:11:12 -0600 5, 23 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 5, 23 -- by mulnx12.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k0OKB14j016596 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 24 Jan 2006 15:11:01 -0500 5, 23 -- Received: from emf03 ([134.53.14.97]) 5, 23 -- by mulnx24.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k0OKB002022369 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 24 Jan 2006 15:11:00 -0500 5, 23 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 5, 23 -- To: microscopy-at-Microscopy.com 5, 23 -- Date: Tue, 24 Jan 2006 15:11:49 -0500 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-type: text/plain; charset=US-ASCII 5, 23 -- Content-transfer-encoding: 7BIT 5, 23 -- Subject: Cryo-ultramicrotomy workshop? 5, 23 -- Message-ID: {43D643B5.14764.643E209-at-localhost} 5, 23 -- Priority: normal 5, 23 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 5, 23 -- X-Real-ConnectIP: 134.53.14.97 5, 23 -- X-Scanned-By: MIMEDefang 2.45 5, 23 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 ==============================End of - Headers==============================
Several years ago, Emispec Systems was purchased by FEI Company of Hillsboro, OR. Recently I've been contacted by several customers who in the past had purchased these data acquisition systems for their EM's and stated they are having difficulty making contact with the responsible person or department at FEI for upgrades and maintenance support.
I'm wondering if parties who are present owners of Emispec ES Vision systems have had success in seeking this support and can provide contact information or advise of appropriate channels to go through to obtain system support.
Please respond off list if you can shed some light on this.
Regards,
Bob Roberts EM Lab Services, Inc. 449 NW 62nd St. Topeka, Kansas 66617-1780 785.246.1232 voice 785.246.0168 fax www.emlabservices.com
==============================Original Headers============================== 8, 23 -- From emlabservices-at-cox.net Tue Jan 24 16:34:05 2006 8, 23 -- Received: from centrmmtao05.cox.net (centrmmtao05.cox.net [70.168.83.79]) 8, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0OMY4Mt004289 8, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 24 Jan 2006 16:34:05 -0600 8, 23 -- Received: from EMLabServices ([24.255.213.54]) by centrmmtao05.cox.net 8, 23 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with SMTP 8, 23 -- id {20060124223407.SQDC5868.centrmmtao05.cox.net-at-EMLabServices} 8, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 24 Jan 2006 17:34:07 -0500 8, 23 -- Message-ID: {012701c62136$49ad42c0$6400a8c0-at-EMLabServices} 8, 23 -- From: "EM Lab Services" {emlabservices-at-cox.net} 8, 23 -- To: {Microscopy-at-microscopy.com} 8, 23 -- Subject: Emispec Systems/FEI Support? 8, 23 -- Date: Tue, 24 Jan 2006 15:34:06 -0700 8, 23 -- MIME-Version: 1.0 8, 23 -- Content-Type: text/plain; 8, 23 -- format=flowed; 8, 23 -- charset="iso-8859-1"; 8, 23 -- reply-type=original 8, 23 -- Content-Transfer-Encoding: 7bit 8, 23 -- X-Priority: 3 8, 23 -- X-MSMail-Priority: Normal 8, 23 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 8, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fmalherbe-at-swin.edu.au) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, January 24, 2006 at 18:23:45 ---------------------------------------------------------------------------
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 sallwein-at-ncifcrf.gov as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Employment Opportunity to work in Support of the National Cancer Institute at SAIC-Frederick, Inc.
Question: SAIC-Frederick, Inc., working in support of the National Cancer Institute at Frederick, seeks an experienced Research Associate to be responsible for independent imaging TEM and SEM samples in a core facility, processing samples and coordinating GLP samples in the lab, processing, sectioning and imaging core samples, training, and conduct collaborative projects with the National Cancer Institute and the National Institutes of Health. Position requires a BS degree and at least four years of experience with TEM and/or SEM sample processing and imaging.
Please visit our web site at http://saic.ncifcrf.gov and refer to opportunity KMB134306 to apply on line.
==============================Original Headers============================== 7, 13 -- From zaluzec-at-microscopy.com Wed Jan 25 07:34:03 2006 7, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 7, 13 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0PDY1FL024339 7, 13 -- for {microscopy-at-microscopy.com} ; Wed, 25 Jan 2006 07:34:03 -0600 7, 13 -- Mime-Version: 1.0 7, 13 -- X-Sender: (Unverified) 7, 13 -- Message-Id: {p06110406bffd2eaa5f0c-at-[206.69.208.22]} 7, 13 -- Date: Wed, 25 Jan 2006 07:34:00 -0600 7, 13 -- To: microscopy-at-microscopy.com 7, 13 -- From: sallwein-at-ncifcrf.gov (by way of MicroscopyListserver) 7, 13 -- Subject: [Filtered] MicroscopyListserverviaWWW: Employment Opportunity to 7, 13 -- work in Support of the National Cancer Institute at SAIC-Frederick 7, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
The 5th International Cryo-EM Course is I believe June 6-17, 2006 in Vancouver, Canada at University of British Columbia by Dr. Elaine Humphrey from UBC and Dr. Kent McDonald, from UC, Berkeley with international speakers. This is an excellent course and caters to the needs of the particular students. Be sure to check with Elaine about dates. For more info email Dr. Elaine Humphrey ech-at-interchange.ubc.ca
Information from the previous course is listed at http://www.emlab.ubc.ca/CryoEM2005/index.html
Judy
Judy Murphy, PhD Microscopy Training, Imaging, and Lab Design Stockton, CA 95219 murphyjudy-at-comcast.net
edelmare-at-muohio.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 14, 19 -- From murphyjudy-at-comcast.net Wed Jan 25 09:51:50 2006 14, 19 -- Received: from sccrmhc14.comcast.net (sccrmhc14.comcast.net [63.240.77.84]) 14, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0PFpnJP012033 14, 19 -- for {microscopy-at-microscopy.com} ; Wed, 25 Jan 2006 09:51:50 -0600 14, 19 -- Received: from [192.168.1.102] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 14, 19 -- by comcast.net (sccrmhc14) with ESMTP 14, 19 -- id {2006012515514401400eiimne} ; Wed, 25 Jan 2006 15:51:44 +0000 14, 19 -- Message-ID: {43D79E99.2010905-at-comcast.net} 14, 19 -- Date: Wed, 25 Jan 2006 07:51:53 -0800 14, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 14, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 14, 19 -- X-Accept-Language: en-us, en 14, 19 -- MIME-Version: 1.0 14, 19 -- To: edelmare-at-muohio.edu, Microscopy List Server {microscopy-at-microscopy.com} 14, 19 -- Subject: Re: [Microscopy] Cryo-ultramicrotomy workshop? 14, 19 -- References: {200601242046.k0OKklJE029987-at-ns.microscopy.com} 14, 19 -- In-Reply-To: {200601242046.k0OKklJE029987-at-ns.microscopy.com} 14, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 14, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Link to current cryocourse June 6-15, 2006 at University of British Columbia, Vancouver, BC Here is the link to the current cryo course in 2006. http://www.emlab.ubc.ca/CryoEM2006/index.html
Judy Murphy wrote:
} The 5th International Cryo-EM Course is I believe June 6-17, 2006 in } Vancouver, Canada at University of British Columbia by Dr. Elaine } Humphrey from UBC and Dr. Kent McDonald, from UC, Berkeley with } international speakers. } This is an excellent course and caters to the needs of the particular } students. Be sure to check with Elaine about dates. } For more info email } Dr. Elaine Humphrey } ech-at-interchange.ubc.ca } } Information from the previous course is listed at } http://www.emlab.ubc.ca/CryoEM2005/index.html } } Judy } } } Judy Murphy, PhD } Microscopy Training, Imaging, and Lab Design } Stockton, CA 95219 } murphyjudy-at-comcast.net } } } } } } } } edelmare-at-muohio.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 had a request from a user looking for a cryo- } } ultramicrotomy workshop. Is anyone offering one in the next year or } } know of one? Thanks! } } } } } } } } Richard E. Edelmann, Ph.D. } } Electron Microscopy Facility Director } } 364 Pearson Hall } } Miami University, Oxford, OH 45056 } } Ph: 513.529.5712 Fax: 513.529.4243 } } E-mail: edelmare-at-muohio.edu } } http://www.emf.muohio.edu } } } } "WE ARE MICROSOFT. } } RESISTANCE IS FUTILE. } } YOU WILL BE ASSIMILATED." } } } } ==============================Original } } Headers============================== } } 5, 23 -- From edelmare-at-muohio.edu Tue Jan 24 14:11:13 2006 } } 5, 23 -- Received: from mulnx12.mcs.muohio.edu } } (mulnx12.mcs.muohio.edu [134.53.6.67]) } } 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } k0OKBBkX020625 } } 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 24 Jan 2006 } } 14:11:12 -0600 } } 5, 23 -- Received: from mulnx24.mcs.muohio.edu } } (mulnx24.mcs.muohio.edu [134.53.6.11]) } } 5, 23 -- by mulnx12.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) } } with ESMTP id k0OKB14j016596 } } 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 24 Jan 2006 } } 15:11:01 -0500 } } 5, 23 -- Received: from emf03 ([134.53.14.97]) } } 5, 23 -- by mulnx24.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) } } with ESMTP id k0OKB002022369 } } 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 24 Jan 2006 } } 15:11:00 -0500 } } 5, 23 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} } } 5, 23 -- To: microscopy-at-Microscopy.com } } 5, 23 -- Date: Tue, 24 Jan 2006 15:11:49 -0500 } } 5, 23 -- MIME-Version: 1.0 } } 5, 23 -- Content-type: text/plain; charset=US-ASCII } } 5, 23 -- Content-transfer-encoding: 7BIT } } 5, 23 -- Subject: Cryo-ultramicrotomy workshop? } } 5, 23 -- Message-ID: {43D643B5.14764.643E209-at-localhost} } } 5, 23 -- Priority: normal } } 5, 23 -- X-mailer: Pegasus Mail for Win32 (v3.12c) } } 5, 23 -- X-Real-ConnectIP: 134.53.14.97 } } 5, 23 -- X-Scanned-By: MIMEDefang 2.45 } } 5, 23 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 } } ==============================End of - } } Headers============================== } } } } } } } }
==============================Original Headers============================== 5, 19 -- From murphyjudy-at-comcast.net Wed Jan 25 09:57:45 2006 5, 19 -- Received: from sccrmhc14.comcast.net (sccrmhc14.comcast.net [204.127.202.59]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0PFviAM012953 5, 19 -- for {microscopy-at-microscopy.com} ; Wed, 25 Jan 2006 09:57:44 -0600 5, 19 -- Received: from [192.168.1.102] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 5, 19 -- by comcast.net (sccrmhc14) with ESMTP 5, 19 -- id {2006012515574101400ejlo9e} ; Wed, 25 Jan 2006 15:57:42 +0000 5, 19 -- Message-ID: {43D79FFF.4030800-at-comcast.net} 5, 19 -- Date: Wed, 25 Jan 2006 07:57:51 -0800 5, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 5, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 5, 19 -- X-Accept-Language: en-us, en 5, 19 -- MIME-Version: 1.0 5, 19 -- To: Microscopy List Server {microscopy-at-microscopy.com} 5, 19 -- Subject: Re: [Microscopy] Cryo-ultramicrotomy workshop? 5, 19 -- References: {200601242046.k0OKklJE029987-at-ns.microscopy.com} {43D79E99.2010905-at-comcast.net} 5, 19 -- In-Reply-To: {43D79E99.2010905-at-comcast.net} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am in need of a manual for the Olympus BX41 microscope, particularly the epifluorescence unit, U-L100HGAPO. If anybody knows of on-line instructions, or how I could obtain a copy of the manuals, I would be very grateful. I will be happy to pay any copying and mailing charges, of course.
Ralph Common Dept. of Physiology Michigan State University
==============================Original Headers============================== 4, 24 -- From rcommon-at-msu.edu Wed Jan 25 10:07:41 2006 4, 24 -- Received: from sys18.mail.msu.edu (sys18.mail.msu.edu [35.9.75.118]) 4, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0PG7dLa015524 4, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Jan 2006 10:07:39 -0600 4, 24 -- Received: from [35.9.122.125] (helo=emlab) 4, 24 -- by sys18.mail.msu.edu with esmtpsa (Exim 4.52 #1) 4, 24 -- (TLSv1:RC4-MD5:128) 4, 24 -- id 1F1nBK-0005iX-UD 4, 24 -- for Microscopy-at-microscopy.com; Wed, 25 Jan 2006 11:07:39 -0500 4, 24 -- From: "Ralph Common" {rcommon-at-msu.edu} 4, 24 -- To: {Microscopy-at-microscopy.com} 4, 24 -- Subject: Olympus BX41 Manual 4, 24 -- Date: Wed, 25 Jan 2006 11:08:14 -0500 4, 24 -- Message-ID: {001201c621c9$8c9bed50$7d7a0923-at-msu.edu} 4, 24 -- MIME-Version: 1.0 4, 24 -- Content-Type: text/plain; 4, 24 -- charset="iso-8859-1" 4, 24 -- Content-Transfer-Encoding: 7bit 4, 24 -- X-Priority: 3 (Normal) 4, 24 -- X-MSMail-Priority: Normal 4, 24 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) 4, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 4, 24 -- Importance: Normal 4, 24 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
Hello Everyone, Is there anyone out there who has experience etching Cu and Ni sulfides, in order to see compositional zoning ?
The phases I am interested in are: Cu2S and Ni3-XS2. In both these compounds Ni and Cu substitute for each other. It is these substitutions which I suspect change the etching behaviour of the compounds. Etching could thus be a quick and dirty alternative to excessive X-ray mapping/analysis.
I don't have the answer, but you may want to post this question on Metallography.com for wider exposure.
Stu Smalinskas, P.E. Metallurgist SKF Plymouth, Michigan
--- shem-at-laurentian.ca wrote: } } Hello Everyone, } Is there anyone out there who has experience etching } Cu and Ni sulfides, in order } to see compositional zoning ? } } The phases I am interested in are: Cu2S and Ni3-XS2. } In both these compounds Ni and Cu substitute } for each other. It is these substitutions which I } suspect change the etching behaviour of the } compounds. } Etching could thus be a quick and dirty alternative } to excessive X-ray mapping/analysis. } } Any input is welcome ! } } Thanks, } } Skage Hem } } } } _______________________________________________________________ } } Skage Hem } Research Scientist, Ph.D. } CAF, Department of Earth Sciences } Laurentian University } Ramsey Lake Road } Sudbury, ON } Canada } P3E 2C6 } } ph. 705-675-1151 x4040 } cell. 705-562-7321 } fax 705-675-4898 } } http://laurentian.ca/geology/Research/hem.html } }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 5, 20 -- From smalinskas-at-yahoo.com Wed Jan 25 15:39:12 2006 5, 20 -- Received: from web34105.mail.mud.yahoo.com (web34105.mail.mud.yahoo.com [66.163.178.103]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k0PLdBtD025507 5, 20 -- for {microscopy-at-sparc5.microscopy.com} ; Wed, 25 Jan 2006 15:39:11 -0600 5, 20 -- Received: (qmail 12361 invoked by uid 60001); 25 Jan 2006 21:39:10 -0000 5, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 20 -- s=s1024; d=yahoo.com; 5, 20 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 5, 20 -- b=1qmKDIUrud/YHnASQmqF6Vva92fi6hEC/jh1UNtRTvJH3OA4jU3Nb6FtrnQAiUR9lMgapzEfGIyQL+xNL6m1zL3xcbR+TJbBeW+komO12e06Smi5V2d6mx9x/L1/xO6qD4YjBOOzBjB4Ck7WEav53M6zgyoDWv+e+oZh+r9g318= ; 5, 20 -- Message-ID: {20060125213910.12359.qmail-at-web34105.mail.mud.yahoo.com} 5, 20 -- Received: from [141.151.33.213] by web34105.mail.mud.yahoo.com via HTTP; Wed, 25 Jan 2006 13:39:10 PST 5, 20 -- Date: Wed, 25 Jan 2006 13:39:10 -0800 (PST) 5, 20 -- From: Kestutis Smalinskas {smalinskas-at-yahoo.com} 5, 20 -- Subject: Re: [Microscopy] Etching of Cu-Ni matte for reflected light microscopy 5, 20 -- To: shem-at-laurentian.ca, 5, 20 -- "microscopy-at-sparc5.microscopy.com" {microscopy-at-ns.microscopy.com} 5, 20 -- In-Reply-To: {200601252114.k0PLE5YS017833-at-ns.microscopy.com} 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; charset=iso-8859-1 5, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
THE TEXAS SOCIETY FOR MICROSCOPY invites you to our Spring 2006 meeting on April 20-22, 2006 at Alcon Research Labs, 6201 South Freeway, Ft. Worth, TX 76134
All registration forms and lodging details are available on our web site: http://www.texasmicroscopy.org/
ABSTRACTS MUST BE RECEIVED BY: March 20, 2006 Advanced Registration Deadline: April 7, 2006. Advanced registration is strongly suggested to afford TSM an accurate participant count for event organization.
**Workshops— Thursday, April 20, 2006 (held at Alcon Labs, Ft. Worth)
“Microwave Immunology 101” Sponsored by Ted Pella, Inc. Speaker: Rick Giberson, Sr. Applications Engineer
“ESEM: not just for Biology Anymore” Sponsored by FEI Company Speaker: Daniel Phifer, Sr. Applications Engineer
**Guest Speaker — Friday, April 21, 2006
“Materials Science in Museums” Dr. Pamela Vandiver, Professor of Materials Science and Engineering and Archeology, Co-Director of Program in Heritage Conservation Science at the University of Arizona, and former Senior Research Scientist at the Smithsonian Institution’s Center for Materials Research and Education.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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Invitation and Call for Papers Contamination Control in Electron and Ion Microscopy Microscopy and Microanalysis 2006, Chicago, IL Organizers: Ronald Vane (XEI Scientific) and András E. Vladár, Ph.D. (NIST) Session A17 (POSTERS ONLY) ABSTRACTS DUE February 15, 2006
This session is being organized to present the latest research on contamination sources and control techniques in charged beam microscopy. Herein we invite papers on all aspects of contamination control inside electron microscopes or focused ion beam tools including sources of contamination, the effects of contamination, contamination artifacts and interference on microscopy results, and contamination control methods and techniques. Contamination includes hydrocarbons, particulates, and other foreign matter that may interfere with imaging and analysis.
Traditionally, contamination control in SEMs has focused on pump oils, fingerprints, dirty specimens, and good vacuum practice in manufacturing and service. The use of dry pumps at all stages of the vacuum system of new FESEMs (Field Emission SEMs) and the use of better vacuum practices on the part on users and manufacturers have reduced contamination, but not eliminated problems. Most commonly, tools in the field begin to exhibit contamination artifacts after several months of use due to trace amounts of contaminants brought in with specimens. The Semiconductor and nano-sciences industries have demanded tools that can image structures {2 nm in size at { 2kV. Instrument manufacturers have responded with Field Emission tools that easily produce better than 400 kX magnification at high contrast with low kV beams. Control of contamination has assumed greater significance as semiconductor companies move to ever smaller dimensions. It is now common to observe features with {2 kV and {10 nm in size, close to these tools’ resolution limits. In such cases, the slightest amount of hydrocarbon (HC) contamination in the chamber can cause loss of resolution and contrast. The electron beam reacts with any stray HC in the beam path or on the surface creating HC ions that condense and form hydrocarbon deposits on the area being scanned. Even with baking, dry pumps and LN traps, artifacts and contamination haze remains.
What are the sources and what can be done? Some Research Topic Ideas:
* Do Hydrocarbons adsorbed on materials migrate on the surface to interact with charged beams to form deposits or is vapor phase transport more important? The effectiveness of cold fingers and Evactron(R) chamber cleaning suggests that vapor phase transport of contaminants is an important alternative mechanism to surface transport.
* Is the gas phase interaction of the electron beam and contaminant molecules important in SEM contamination buildup? Electron impact ionization is a common technique in mass spectrometry of organics. What is the importance of electron impact ionization on vapor phase organics in the SEM in causing contamination build up in scanned areas?
* What gives better results: plasma cleaning every specimen before introduction to the SEM chamber, Evactron chamber cleaning, or Evactron cleaning specimens in the chamber?
* Study Hydrocarbon removal rates for different decontamination methods, for different types of hydrocarbons and concentrations, under different Evactron De-Contaminator power and pressure settings, and for different pump speeds and flow rates during Evactron cleaning.
* How fast does AMC (atmospheric molecular contamination) accumulate on specimens in different environments after they are cleaned? Freshly prepared and cleaned specimens are known to be freer from contamination than specimens that have sat in room air for several days. Quantify this effect.
* Establish a standard procedure for depositing repeatable amounts of contamination for removal technique studies.
* Establish a protocol for measuring tool contamination that can be transferred between tools and used to compare tools or used to establish contamination standards or limits.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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Email: kjl226-at-vt.edu Name: Kathy
Organization: Virginia Tech
Title-Subject: [Filtered] Embedding spores
Question: What is the best way to process and embed spores?
Why not just ask Olympus, I'm pretty sure they will have one as its still a current microscope. If you are happy with a photocopy expect to get it free of charge from your local rep (and I'm sure they will email you the pdf version as well).
Zeiss and Leica now have all the recent microscope manuals on-line, but I have obtained photocopied manuals for 20+ year old microscopes and fittings, and once even a BASIC program hardware/software guide for a very odd programmable HP+Zeiss motorised XY stage discontinued in the 1970's (in the 1990's). The most interesting bit was the BASIC 'Trace on' debugging command: 'TRON', but to be honest it worked better when Disney did it.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
The session organizers for the Pharmaceutical Symposium of M&M 2006 are looking for additional speakers. Presentations should discuss biological or material science applications of significance to the pharmaceutical industry. The meeting will be held in Chicago, July 30 - Aug. 3. The abstract submission deadline is Feb. 15, 2006. If you are interested in submitting a platform or poster presentation, instructions for abstract submission are available at the meeting web site at http://mm2006.microscopy.org/ . For additional questions please contact the session chairs at the addresses below. Thanks.
Joe Neilly Abbott Laboratories 200 Abbott Park Rd. Bldg. AP31, Dept. R4R9 Abbott Park, IL 60064-6202 email: joe.neilly-at-abbott.com voice: 847-938-5024
Jim DiOreo Baxter Healthcare RL WG3-2S Route 120 and Wilson Road Round Lake, IL 60002 email: jim_dioreo-at-baxter.com voice: 847-270-4676
==============================Original Headers============================== 3, 18 -- From joe.p.neilly-at-abbott.com Thu Jan 26 07:59:45 2006 3, 18 -- Received: from abtmx5.abbott.com (abtmx5.abbott.com [130.36.44.95]) 3, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0QDxiEW028526 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 26 Jan 2006 07:59:45 -0600 3, 18 -- Received: from abtapn561.northamerica.intra.abbott.com (abtapn561.northamerica.intra.abbott.com [10.236.188.103]) 3, 18 -- by abtmx5.abbott.com (Switch-3.1.7/Switch-3.1.7) with ESMTP id k0QDxeU7010209 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 26 Jan 2006 07:59:40 -0600 (CST) 3, 18 -- To: microscopy-at-microscopy.com 3, 18 -- Subject: Request for Speakers at Pharmaceutical Symposium , M&M 2006 3, 18 -- X-Mailer: Lotus Notes Release 5.0.5 September 22, 2000 3, 18 -- Message-ID: {OFE0F856E6.F3ED62DE-ON86257102.004CC6F7-at-abbott.com} 3, 18 -- From: Joseph P Neilly {joe.p.neilly-at-abbott.com} 3, 18 -- Date: Thu, 26 Jan 2006 07:59:12 -0600 3, 18 -- X-MIMETrack: Serialize by Router on ABTAPN561/ESVR/ABBOTT(Release 6.5.4FP2|September 12, 2005) at 3, 18 -- 01/26/2006 07:59:42, 3, 18 -- Serialize complete at 01/26/2006 07:59:42 3, 18 -- MIME-Version: 1.0 3, 18 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
We have a networked color laser printer in our lab which we use to print routine image / working print from our SEM's and otehr microscopes. We print images and sectra, EBDS, etc. What we have found is that fewer and fewer folks actually want hardcopies. They want and use electronic versions, since even for publication nearly all of the journals these days want electronic only. My printer supplies are about 20% or lower of what they were 2-3 years ago (i.e. over an 80% decrease in printing - at the same time I've seen a 28% increase in user numbers).
For high quality and archive prints we use our Epson 9600 wide format printer - we generally use if for posters. I would not hesitate to get another Epson Pro series, perhaps in a smaller format for more routine size prints. Or look at the HP highend printers.
But remember the inks will dry in the printers if they are not regularly used. Some of the manufacturers have inks which are designed to last longer in the printer - but they are not the low- to medium end printers. Dry inks like laser printers and wax/polymer "phaser" printers have an advantage here.
FYI: We have a Lexmark C752 color laser printer. And whereas we've been very very happy with the durablity and quality of the Lexmark Black / Greyscale laser printers, we have not been thrilled with the image quality of the C752 prints, and yes we use the high quality color laser paper.
On 25 Jan 2006, at 7:44, fmalherbe-at-swin.edu.au wrote:
} } } } ---------------------------------------------------------------------- } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (fmalherbe-at-swin.edu.au) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Tuesday, January 24, 2006 at 18:23:45 } ---------------------------------------------------------------------- } ----- } } Email: fmalherbe-at-swin.edu.au } Name: Francois Malherbe } } Organization: Swinburne University of Technology } } Education: Graduate College } } Location: Melbourne, Victoria, Australia } } Question: We are purchasing a new SEM, what are the basic specs you } would recommend for the printer: } } - inkjet, laser or other? } - USB, serial? } - dpi? } } Thank you for your help. } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== 9, 12 -- From } zaluzec-at-ultra5.microscopy.com Wed Jan 25 07:33:17 2006 9, 12 -- } Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 9, } 12 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k0PDXFin024204 9, 12 -- for {microscopy-at-microscopy.com} ; Wed, 25 Jan } 2006 07:33:16 -0600 9, 12 -- Mime-Version: 1.0 9, 12 -- X-Sender: } zaluzec-at-ultra5.microscopy.com (Unverified) 9, 12 -- Message-Id: } {p06110405bffd2e8756cc-at-[206.69.208.22]} 9, 12 -- Date: Wed, 25 Jan } 2006 07:33:15 -0600 9, 12 -- To: microscopy-at-microscopy.com 9, 12 -- } From: fmalherbe-at-swin.edu.au (by way of Ask-A-Microscopist) 9, 12 -- } Subject: [Filtered] AskAMicroscopist: Printer for an SEM 9, 12 -- } Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
==============================Original Headers============================== 12, 25 -- From edelmare-at-muohio.edu Thu Jan 26 09:13:08 2006 12, 25 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 12, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0QFD7wO013454 12, 25 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Jan 2006 09:13:07 -0600 12, 25 -- Received: from mulnx23.mcs.muohio.edu (mulnx23.mcs.muohio.edu [134.53.6.10]) 12, 25 -- by mulnx11.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k0QFD5ac030704; 12, 25 -- Thu, 26 Jan 2006 10:13:05 -0500 12, 25 -- Received: from emf03 ([134.53.14.97]) 12, 25 -- by mulnx23.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k0QFD4fg029547; 12, 25 -- Thu, 26 Jan 2006 10:13:04 -0500 12, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 12, 25 -- To: fmalherbe-at-swin.edu.au 12, 25 -- Date: Thu, 26 Jan 2006 10:13:59 -0500 12, 25 -- MIME-Version: 1.0 12, 25 -- Content-type: text/plain; charset=US-ASCII 12, 25 -- Content-transfer-encoding: 7BIT 12, 25 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: Printer for an SEM 12, 25 -- CC: microscopy-at-Microscopy.com 12, 25 -- Message-ID: {43D8A0E7.2878.F7FC50B-at-localhost} 12, 25 -- Priority: normal 12, 25 -- In-reply-to: {200601251344.k0PDiMAx027101-at-ns.microscopy.com} 12, 25 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 12, 25 -- X-Real-ConnectIP: 134.53.14.97 12, 25 -- X-Scanned-By: MIMEDefang 2.45 12, 25 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.10 ==============================End of - Headers==============================
Do you mean using glass plates instead of film? If so ..... Plates are much thicker and heavier, these factors contribute to storage problems. Plates break when dropped, film does not. I know people who insist that plates give a better image but I have seen no evidence of that. The boxes that Kodak plates came in were great for storing lantern slides.
Geoff
Ken.Blight-at-cancer.org.uk wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 8, 32 -- From mcauliff-at-umdnj.edu Thu Jan 26 09:19:07 2006 8, 32 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 8, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0QFJ71O014621 8, 32 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Jan 2006 09:19:07 -0600 8, 32 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 8, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 263D14BE18 8, 32 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Jan 2006 10:19:07 -0500 (EST) 8, 32 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 8, 32 -- by zix03.umdnj.edu (Proprietary) with ESMTP id 785834BDF9 8, 32 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Jan 2006 10:19:06 -0500 (EST) 8, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 32 -- id {0ITP00L01GB01W-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 32 -- for microscopy-at-msa.microscopy.com; Thu, 26 Jan 2006 10:19:06 -0500 (EST) 8, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 8, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 32 -- 2004)) with ESMTP id {0ITP00JRVGQKBF-at-Polaris.umdnj.edu} ; Thu, 8, 32 -- 26 Jan 2006 10:08:58 -0500 (EST) 8, 32 -- Date: Thu, 26 Jan 2006 10:08:10 -0500 8, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 32 -- Subject: Re: plates 8, 32 -- In-reply-to: {200601261358.k0QDwKjM028252-at-ns.microscopy.com} 8, 32 -- To: Ken.Blight-at-cancer.org.uk, 8, 32 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 32 -- Message-id: {43D8E5DA.6030702-at-umdnj.edu} 8, 32 -- MIME-version: 1.0 8, 32 -- Content-type: text/plain; format=flowed; charset=us-ascii 8, 32 -- Content-transfer-encoding: 7BIT 8, 32 -- X-Accept-Language: en-us, en 8, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 32 -- Gecko/20040804 Netscape/7.2 (ax) 8, 32 -- References: {200601261358.k0QDwKjM028252-at-ns.microscopy.com} ==============================End of - Headers==============================
Sorry for the confusion caused -The imaging plates I am interested in are electron detection plates for high quality digital images. I have no other details otherwise I would not be asking the question!
ken
Ken Blight Senior Scientific Officer Electron Microscopy Cancer Research UK London England.
==============================Original Headers============================== 4, 14 -- From Ken.Blight-at-cancer.org.uk Thu Jan 26 09:29:15 2006 4, 14 -- Received: from magic.lif.icnet.uk (magic.lif.icnet.uk [143.65.100.6]) 4, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0QFTEVw017335 4, 14 -- for {microscopy-at-microscopy.com} ; Thu, 26 Jan 2006 09:29:14 -0600 4, 14 -- Received: from [143.65.59.12] ([143.65.59.12]) 4, 14 -- by magic.lif.icnet.uk (8.9.3p2/8.9.3) with ESMTP id PAA27526 4, 14 -- for {microscopy-at-microscopy.com} ; Thu, 26 Jan 2006 15:29:13 GMT 4, 14 -- Mime-Version: 1.0 4, 14 -- Message-Id: {a06200702bffe98674b18-at-[143.65.59.12]} 4, 14 -- Date: Thu, 26 Jan 2006 15:25:01 +0000 4, 14 -- To: microscopy-at-microscopy.com 4, 14 -- From: Ken Blight {Ken.Blight-at-cancer.org.uk} 4, 14 -- Subject: Imaging plates 4, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
What you want is described at http://www.berthold.com.au/imaging_pages/FDL-5000.html
and at http://www.ditabis.de/
I have no experience with use of these in EM. An impressive 3D cryo-EM paper based on use of the imaging plates was published by Holmes et al in Nature (2003), 425:423-427. Use of imaging plates requires drying the media before loading into the camera, and requires breaking camera vacuum and accessing the camera in order to remove media to "develop" (scan & digitize) the recorded images and regenerate the plate. Both steps are avoided when using CCD display and recording of EM images, which provide the other route for directly digitizing EM images without processing and scanning photographic negatives.
-mike reedy-
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-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
I hope you all will forgive this detour from microscopy. If you have a x-ray film processor in your lab and would care to comment on X-ray film processors I would love to hear from you off-list. We have an AFP Medical-Mini X-ray film processing system but we are looking into replacing it with a Konica SRX-101A desktop processor. Advice/ opinions on this equipment would be greatly appreciated. Thanks, 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)
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Title-Subject: [Filtered] TEM - Seeking to acquire JEOL jem100c or equivalent.
Question: We are a small French company based in the Paris region, currently looking to replace our JEOL jem100c transmission electron microscope, which unfortunately, has broken down.
We need to acquire a second-hand microscope similar to the model we actually have but as we wish to use it mainly as an electron generator, to test the performance of the customized cameras we manufacture, we would accept a tem microscope, which does not offer all the requirements necessary for good quality imagery.
If you have such equipment and are willing to give us the opportunity of acquiring it, please contact us via our e-mail address: info-at-lheritier-sa.com.
It looks like we are losing the +24V section of the power supply on our Oxford ISIS EDS. Does anyone have a spare power supply, or even an entire ISIS unit they would be willing to part with? It probably should be a model 200 or 300.
Please contact me off-line and we can discuss the details.
Warren Straszheim Materials Analysis and Research Lab Iowa State University 515-294-8187
==============================Original Headers============================== 4, 29 -- From wesaia-at-iastate.edu Fri Jan 27 13:37:34 2006 4, 29 -- Received: from mailhub-5.iastate.edu (mailhub-5.iastate.edu [129.186.140.15]) 4, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0RJbXOC001093 4, 29 -- for {Microscopy-at-Microscopy.Com} ; Fri, 27 Jan 2006 13:37:34 -0600 4, 29 -- Received: from mailout-2.iastate.edu (mailout-2.iastate.edu [129.186.140.2]) 4, 29 -- by mailhub-5.iastate.edu (8.12.11/8.12.10) with SMTP id k0RJbWHI009451 4, 29 -- for {Microscopy-at-Microscopy.Com} ; Fri, 27 Jan 2006 13:37:32 -0600 4, 29 -- Received: from owa.eng.iastate.edu(129.186.23.85) by mailout-2.iastate.edu via smtp 4, 29 -- id 7bca_5f49d450_8f6c_11da_9c60_003048290bef; 4, 29 -- Fri, 27 Jan 2006 13:37:37 -0600 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, 27 Jan 2006 13:37:32 -0600 4, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 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: Need ISIS power supply 4, 29 -- Date: Fri, 27 Jan 2006 13:38:47 -0600 4, 29 -- Message-ID: {16A330AC32056A40B32842EC4BB8D7275F1FFC-at-maire.eng.iastate.edu} 4, 29 -- X-MS-Has-Attach: 4, 29 -- X-MS-TNEF-Correlator: 4, 29 -- Thread-Topic: Need ISIS power supply 4, 29 -- Thread-Index: AcYjeUgRcQ/G34hJQ8agDRbdnqwxXA== 4, 29 -- From: "Straszheim, Warren E [CCE E]" {wesaia-at-iastate.edu} 4, 29 -- To: {Microscopy-at-Microscopy.Com} 4, 29 -- X-OriginalArrivalTime: 27 Jan 2006 19:37:33.0133 (UTC) FILETIME=[1E3347D0:01C62379] 4, 29 -- Content-Transfer-Encoding: 8bit 4, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0RJbXOC001093 ==============================End of - Headers==============================
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Question: Title: Advanced Techniques in Microscopy for Materials Characterization
Lecturers: David Joy, University of Tennessee Eric Lifshin, State University of New York George Vander Voort, Buehler Ltd. Raynald Gauvin, McGill University Brendan Griffin, University of Western Australia Rocco Cerchiara, Fischione Instruments Pierre Hovington, Hydro-Quebec Research Institute Tom Kelly, Imago Scientific Instruments Scott Sitzman, HKL Technology Marin Lagace, Hydro-Quebec Research Institute
When: May 8-12, 2006
Where: McGill University, Department of Mining, Metals and Materials Engineering M.H.Wong Building 3610 University Street Montreal, Quebec, Canada H3A 2B2
The worlds largest and most comprehensive microscopy and microanalysis meeting is coming to Chicago July 30-Aug 3, 2006. This is the perfect opportunity for both your and as well as undergraduate/graduate students to gain valuable insights to recent work by interacting with the worlds most prominent researchers, as well as learn about the latest techniques in the scientific symposia and in instrumentation.
In addition to more than 50 scientific sessions and symposia there will be a number of short courses/tutorials starting on the weekend and extending through the week.
The meeting also hosts the largest commerical exhibition of microscopy instrumentation at any venue.
This will be the perfect opportunity for you to take advantage of the premier international meeting happening in the USA as well as make valuable contacts for the future.
Of particular importance to students is the availability of scholarship, and other funding opportunities for students which will allow you to attend the meeting at reduced costs.
*Student receive reduced registration rates * Scholarships and Awards are available from the sponsoring societies * Poster awards are presented for the best student contributions * Student Bursaries (i.e. work part time during the meeting) can be used to help defray your expenses
A limited number of awards are also available for professional technical staff.
To be eligible for these opportunities you must submit a paper for either platform or poster presentation at the meeting. The deadline for submitting a paper to MM2006 is Feb. 15, 2006 (there are no extensions), which is only a few weeks away.
If you have any question please feel free to contact any of the Organizers, for detailed information please visit the meeting WWW site (http://mm2006.microscopy.org).
Again the paper submission deadline is Feb 15, 2006. I will also point out that the submission process is now entirely electronic so you still have plenty of time to prepare your work and submit it for review and consideration. All papers for accepted for the meeting are included in the proceedings which is published as supplement to the Journal of the Microscopy Society of America - Microscopy & Microanalysis.
Disclaimer: In additions to being your Friendly Neighborhood SysOp, I have the "honor" (cough) of being the Co-chair of the local arrangements committee for this meeting, thus I have a vested interest in getting you to come to Chicago!
Cheers...
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 16, 11 -- From zaluzec-at-microscopy.com Mon Jan 30 08:31:29 2006 16, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 16, 11 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0UEVSk1010268 16, 11 -- for {microscopy-at-microscopy.com} ; Mon, 30 Jan 2006 08:31:29 -0600 16, 11 -- Mime-Version: 1.0 16, 11 -- Message-Id: {p06110401c003ccb17092-at-[206.69.208.22]} 16, 11 -- Date: Mon, 30 Jan 2006 08:31:28 -0600 16, 11 -- To: microscopy-at-microscopy.com 16, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 16, 11 -- Subject: MM 2006 Paper Submission Deadline - Feb 15, 2006 16, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Hi Everyone I am looking for a EHT X-regulator for our old Phillips EM 300 TEM. Anybody knows a good place to buy this part or have such parts on sale please contact me at pjoshi-at-miami.edu Thank you
Pratik P. Joshi, Ph.D.
Assistant Scientist / Lab Manager Center for Advanced Microscopy Department of Chemistry, University of Miami 1301 Memorial Drive, Room 315 Miami, FL- 33146 Ph: (305)284-4736, Fax: (305)284-1880
==============================Original Headers============================== 2, 20 -- From pjoshi-at-miami.edu Mon Jan 30 08:58:40 2006 2, 20 -- Received: from EXCHANGE2.cgcent.miami.edu (ntexchsrv2.ir.miami.edu [129.171.32.59]) 2, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0UEwd88014815 2, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 30 Jan 2006 08:58:39 -0600 2, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 2, 20 -- Content-class: urn:content-classes:message 2, 20 -- MIME-Version: 1.0 2, 20 -- Content-Type: text/plain; 2, 20 -- charset="iso-8859-1" 2, 20 -- Subject: Re: Looking for EHT X Regulator unit for Phillips EM 300 TEM 2, 20 -- Date: Mon, 30 Jan 2006 09:58:34 -0500 2, 20 -- Message-ID: {B2BA593DA3EBE143A856ABA856D01F5D02DCFE55-at-EXCHANGE2.cgcent.miami.edu} 2, 20 -- X-MS-Has-Attach: 2, 20 -- X-MS-TNEF-Correlator: 2, 20 -- Thread-Topic: Looking for EHT X Regulator unit for Phillips EM 300 TEM 2, 20 -- Thread-Index: AcYlraSDK/Vfxr9oTDWLGaysH9W/UA== 2, 20 -- From: "Joshi, Pratik P" {pjoshi-at-miami.edu} 2, 20 -- To: {Microscopy-at-microscopy.com} 2, 20 -- Content-Transfer-Encoding: 8bit 2, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0UEwd88014815 ==============================End of - Headers==============================
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Email: brad.huggins-at-bp.com Name: Brad Huggins
Organization: BP Chemicals, Naperville, IL
Title-Subject: [Filtered] LM, Anyone with "Digital Microscope" familiarity/experience?
Question: (trouble with MS Outlook and spam errors, so I'm using this format)
For several remote, light microscope applications, I have been looking at the Keyence VHX-100, Digital Microscope. It looks to be a very capable and versatile tool. Aside from its portability, 3D imaging capability appears quite impressive to me. It looks capable of adding - very significantly - to the limited depth-of-focus imaging condition of our existing light microscope systems.
Does anyone on the ListServer have any experience (good, bad or otherwise) with this microscope? Or similar digital microscopes? If you would rather not respond online, please share with me off-line. I will share a summary of "anonymous responses" as appropriate.
Thanks in advance,
Brad Huggins
BP Chemicals Naperville, IL brad.huggins-at-bp.com 630 420-3668
I would like hear the comment on various sample preparation technique for confocal microscopy (indirect Immunofluorescence) which gives 1) best presarvation 2) minimum background noise
Is it advisible to dry the slide before switching over to 1 Ab and 2Ab. I mean to ask is this drying in between lead to some form of precipitate???
thanks in advance
Regards, Shrunali Kulkarni Scientist Institute of Microbial Technology India
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 6, 18 -- From aarti_harle-at-yahoo.co.in Tue Jan 31 22:35:20 2006 6, 18 -- Received: from web8303.mail.in.yahoo.com (web8303.mail.in.yahoo.com [202.43.219.124]) 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k114ZGAT025969 6, 18 -- for {Microscopy-at-microscopy.com} ; Tue, 31 Jan 2006 22:35:17 -0600 6, 18 -- Received: (qmail 86863 invoked by uid 60001); 1 Feb 2006 04:35:18 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.co.in; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 18 -- b=AkdqGUkec0jNq9kRo7Ys1VDUDL9lmWiPYTrbZimG2x1gKtvKw56/ZRooXkIC5egeuXa/ocAJv2H7KBo8JPcd2YKl7to7s51Bgw1hKsKk03kglfqBuZIch4AbZnTxASV5UcSCJ1oiDR9TfNDpAWfoQmdp+Ov+/Y2ZET4xGUPfwjE= ; 6, 18 -- Message-ID: {20060201043518.86861.qmail-at-web8303.mail.in.yahoo.com} 6, 18 -- Received: from [203.197.210.215] by web8303.mail.in.yahoo.com via HTTP; Tue, 31 Jan 2006 20:35:18 PST 6, 18 -- Date: Tue, 31 Jan 2006 20:35:18 -0800 (PST) 6, 18 -- From: Aarti Harle {aarti_harle-at-yahoo.co.in} 6, 18 -- Subject: Sample preparation technique for confocal microscopy 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==============================
Yes it works fine (as do many anti-fadents e.g Citifluor http://www.citifluor.co.uk ), although we rarely get problems with DAPI bleaching - I won't mention Hoescht as I can never spell it. Normally the likes of Cy5 or TRITC type fluorochromes bleach first, and anyway DAPI is normally only there in our case to identify the animal cells more easily (and it really makes specimen focussing a cinch with our 'low contrast' samples). Vectashield antifadents stop samples fading really quickly rather than eliminating fading. So you still have to minimise laser (and Hg lamp) exposure times (e.g. increase scan speed, reduce image size & image averaging, increase gain, and, normally in the last resort, open up the confocal iris), particularly if you are doing Z stacks (naturally time-lapse won't be a problem with fixed 'Vectashield' samples).
Being Cell Biology, most of our specimens are derived from cell cultures and so have little contrast, making cell visual separation and location more difficult even with DIC. We only have a little 5 Mw 'violet' 405nm laser with our Leica SP2 AOBS confocal, but it works very well with nuclear stains despite being at the very edge of the DAPI/Hoescht excitation spectrum. We mostly use Mattek dishes (Petri dishes with a hole at the base and little cover slips glued on - see http://www.glass-bottom-dishes.com/ ) as all our microscopes are inverted and we only use high power oil objectives. They have the advantage that we can simply drip the Vectashield into the Petri dish on top of the fixed cells and replenish if it dries out (samples often keep for days or longer in a darkened fridge). With glass slides, the coverslip needs sealing on over the Vectashield with nail varnish, but the nail varnish (and marker pen ink) often dissolve into the immersion oil making a bit of a mess, particularly with the inverted microscopes we use. The nail varnish also sticks to the stage slide holder (as the slide has to be placed upside down in inverted systems) making a sticky mess and knocking the sample out of level.
With regard to preparation, we don't deal with microbiological specimens, just mammalian cells - so I won't offer any advise. There are plenty of recipes on the internet and in books though, and you can try contacting staff at another microbiological institutes via on-line searches or traditional 'old boy' networks. This list server tends to be biased a little towards electron microscopy. You reduce confocal image 'noise' in samples by lowering the scan speed, increasing image averaging and upping laser power on the confocal - the exact opposite of that required to minimise beaching. So there is always a compromise between image quality and fluorochrome bleaching. If the sample preparation has worked well your sample would be expected to be quite bright initially (prior to bleaching) - often problems with dark images are due to experimental failure rather than the confocal microscope and it's software.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {aarti_harle-at-yahoo.co.in} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, January 31, 2006 1:02 PM
Dear Shrunali,
I forgot to add it (although I expect you have the link already) : The full details on Vectorshield products [e.g. Hardset and Mounting Medium versions] can be found on the manufacturers website :
It naturally has the 'test on an inconspicuous area before using' disclaimer. Anti-fadents have been reported to occasionally cause a leaching of stain and loss of fluorescence in some instances.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {aarti_harle-at-yahoo.co.in} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, January 31, 2006 1:02 PM
I am interested in hearing what type of gases people are using in their ESEM applications. We recently learned our silicon drift detector is incompatible with water vapor, and we apparently cannot complete any x-ray microanalysis while working in standard ESEM or VP modes (I was pretty surprised to learn this). We are thinking that using a dry gas such as nitrogen or helium will allow us to work in ESEM modes and use our x-ray system as well, as there is no potential for moisture condensing on the crystal surface.
Our ESEM is a Quanta 200, and it is not equipped with the peltier stage, so we mainly use the environmental modes to alleviate charging in uncoated samples. The x-ray system was purchased with the microscope about four years ago. I would rather not discuss the name of the x-ray vendor on the list, as we have a significant investment in this detector and do not foresee finding the money to purchase a new one soon.
Our first concern is the impact of the gas on spectra. We can coat samples and run them in high vacuum mode, but customers like spectra that represent the sample and not the coating material. We periodically have samples that cannot be coated, so ESEM becomes critical for our imaging needs. If the gas is going to have a dramatic impact on signal, are we better off coating and running high vacuum?
Is there a best choice for chamber gas? Our service engineer recommends nitrogen as having benefits for keeping the system clean. We can set it up fairly quickly, and I have a spare tank and regulator. I have heard of people using helium and believe that I read somewhere that there was some advantage to using helium.
I am still trying to get information from the vendor on whether this will allow us to use the x-ray and ESEM simultaneously, or whether we have options with different collimators or windows. Their responses have been pretty slow coming.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238
==============================Original Headers============================== 9, 23 -- From hagglundk1-at-nku.edu Wed Feb 1 08:09:02 2006 9, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 9, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k11E92h6029071 9, 23 -- for {microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 08:09:02 -0600 9, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 9, 23 -- Wed, 1 Feb 2006 09:09:02 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 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: ESEM chamber gas choices 9, 23 -- Date: Wed, 1 Feb 2006 09:09:01 -0500 9, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F35861A-at-mailfac1.hh.nku.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: ESEM chamber gas choices 9, 23 -- Thread-Index: AcYnOQywNZQdWqyLTzaT/PcS/myvmA== 9, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 01 Feb 2006 14:09:02.0302 (UTC) FILETIME=[0DB1E7E0:01C62739] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k11E92h6029071 ==============================End of - Headers==============================
I am looking for information about staffing, equipment and general practices in microscopy facilities at other institutions. If anyone else is interested in the same sort of information, please reply to me directly. Thank you.
Lesley Bechtold
Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322
==============================Original Headers============================== 7, 21 -- From lesley.bechtold-at-jax.org Wed Feb 1 09:23:28 2006 7, 21 -- Received: from carmen.jax.org (carmen.jax.org [192.43.249.16]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k11FNO0c004460 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 09:23:26 -0600 7, 21 -- Received: from jcs-mid-prod.jax.org (jcs-mid-prod.jax.org [192.43.249.134]) 7, 21 -- by carmen.jax.org (8.12.11/8.12.11) with ESMTP id k11FLEmh001364 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 10:23:14 -0500 (EST) 7, 21 -- (envelope-from lesley.bechtold-at-jax.org) 7, 21 -- Received: from spikey.jax.org by jcs-mid-prod.jax.org 7, 21 -- with ESMTP id 65124051138807281; Wed, 01 Feb 2006 10:21:21 -0500 7, 21 -- From: "Lesley Bechtold" {lesley.bechtold-at-jax.org} 7, 21 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} 7, 21 -- Subject: Seeking information 7, 21 -- Date: Wed, 1 Feb 2006 10:20:25 -0500 7, 21 -- Message-ID: {20060201102025314.00000001972-at-spikey} 7, 21 -- X-Mailer: Oracle Connector for Outlook 10.1.1.0.5 71011 (11.0.6568) 7, 21 -- X-Accept-Language: en-us, en 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; charset=us-ascii 7, 21 -- Content-Transfer-Encoding: 8bit 7, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k11FNO0c004460 ==============================End of - Headers==============================
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Email: ptomic-at-ciclonsemi.com Name: Peter Tomic
Organization: Ciclon Semiconductor Device Corp.
Title-Subject: [Filtered] ELYMA
Question: I would like to hear from people in the semiconductor industry on their experience with ELYMAT [Electrolytical Metal Analysis Tool]. This is an imaging system for the analysis of metal and oxygen contamination in Si wafer technology.
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Email: twigg-at-estd.nrl.navy.mil Name: Mark Twigg
Organization: Naval Research Laboratory
Title-Subject: [Filtered] Postdoctural Position
Question: Postdoctoral Research Position Electronics Science and Technology Division Naval Research Laboratory, Washington, DC
We are seeking a Postdoctoral fellow with a strong transmission electron microscopy and extended-defects/nanostructures background to join our electronic materials program at NRL. You will utilize your skills to characterize SiC and GaN epitaxial layers, as well as nanoelectronic materials based on colloidal gold. We have our own Hitachi H9000UHR HRTEM and sample preparation laboratory. We also have access to a Philips CM30, a JEOL 2200 FETEM with Omega energy filter and Z-contrast STEM, and an FEI Nova 600 Dual Beam FIB. The qualified candidate should have a Ph.D. in Materials Science, Physics, Chemistry, or a related physical science or engineering discipline. Experience in a variety of electron microscopy techniques including HRTEM, CBED, and EDXS, and EELS is important. Experience with FIB, XTEM, and mechanical lapping sample preparation techniques is also desirable. Participants must be citizens or permanent residents of the United States. Two postdoctoral study programs are available at the Naval Research Laboratory: NRC and ASEE. NRC postdoctoral positions are administered by the National Research Council, a division of the National Academy of Sciences. The ASEE Postdoctoral Fellowship Program is administered by the American Society for Engineering Education. Further information about NRL's NRC post-doc program is found on the NRC web site (http://hroffice.nrl.navy.mil/jobs/postdoc.htm). Fellowships are awarded for one year and may be extended for a second year. The base annual stipend for both programs the first year is $62,886.
Point of Contact: Dr. Mark E. Twigg Naval Research Laboratory Code 6812 4555 Overlook Avenue, S.W. Washington, DC 20375-5320
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Has anyone used this buffer before? I plan on using this to make up a fixative to perfuse fix a rat. I will be looking at myelin of the sciatic nerve. Why does this buffer have such a high osmolarity? Any thoughts?
Chip, I've never used Tyrode's in conjunction with cacodylate. Years ago, when the lab I was in was collaborating with an electrophysiologist looking at structure-function relationships in heart muscle, we used Tyrode's because it is a bicarbonate-buffered solution and he could keep the isolated papillary muscles happy in it for hours as long as he bubbled oxygen-CO2 through it. Lovely structure (see various papers by Robinson, TR, etal during the 1980's). 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
==============================Original Headers============================== 1, 21 -- From lcgould-at-med.cornell.edu Wed Feb 1 12:54:15 2006 1, 21 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k11IsEXQ002540 1, 21 -- for {microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 12:54:15 -0600 1, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 21 -- by smtp-gw2.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k11IsBig011013 1, 21 -- for {microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 13:54:11 -0500 (EST) 1, 21 -- Received: from [140.251.48.23] by mpx1.med.cornell.edu 1, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 21 -- with ESMTP id {0IU0000UJV6ABKC0-at-mpx1.med.cornell.edu} for 1, 21 -- microscopy-at-microscopy.com; Wed, 01 Feb 2006 13:54:11 -0500 (EST) 1, 21 -- Date: Wed, 01 Feb 2006 13:50:59 -0500 1, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 21 -- Subject: Re: [Microscopy] viaWWW: Tyrodes-Cacodylate Buffer 1, 21 -- In-reply-to: {200602011727.k11HRkBD005210-at-ns.microscopy.com} 1, 21 -- To: dyel-at-mail.nih.gov, microscopy-at-microscopy.com 1, 21 -- Message-id: {p06200705c006b2a8a260-at-[140.251.48.23]} 1, 21 -- MIME-version: 1.0 1, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 21 -- References: {200602011727.k11HRkBD005210-at-ns.microscopy.com} 1, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.02.01.100606 ==============================End of - Headers==============================
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Has anyone used this buffer before? I plan on using this to make up a fixative to perfuse fix a rat. I will be looking at myelin of the sciatic nerve. Why does this buffer have such a high osmolarity? Any thoughts?
Pan American Advanced Studies Institute on Transmission Electron Microscopy in Materials Science
July 9 to July 22, 2006
Expenses paid Further information: http://www.pasi-tem2006.cl
At the University of Chile - in Santiago, Chile - a new field-emission TEM is being installed. This microscope will form the basis of a Pan American Advanced Studies Institute which will cover principles and applications of TEM to topics in materials and geological sciences. The Institute is funded by the National Science Foundation of the United States.
We invite applications from students and young researchers who wish to participate in this two-week intensive workshop. APPLICANTS WHO ARE ACCEPTED WILL HAVE THEIR EXPENSES PAID. The number of participants in the workshop will be limited to 48.
The lectures and laboratories (presented by a distinguished group of lecturers) will treat a wide range of topics: principles and applications of transmission electron microscopy; microanalysis; latest results and advances; and sample preparation.
The Institute is open to participants from any country in the Americas (including the U.S.A.). Participation by women and members of minority groups is greatly encouraged. Details of how to apply are given on the web site: http://www.pasi-tem2006.cl The closing date for applications is March 13, 2006
-- ........... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
==============================Original Headers============================== 10, 22 -- From jae5-at-lehigh.edu Wed Feb 1 15:13:05 2006 10, 22 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 10, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k11LD4QL023398 10, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 15:13:04 -0600 10, 22 -- Received: from [128.180.55.141] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 10, 22 -- (authenticated bits=0) 10, 22 -- by rain.CC.Lehigh.EDU (8.13.5/8.13.5) with ESMTP id k11LD3Wn022378 10, 22 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 10, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 16:13:03 -0500 10, 22 -- Message-ID: {43E1245F.1070609-at-lehigh.edu} 10, 22 -- Date: Wed, 01 Feb 2006 16:13:03 -0500 10, 22 -- From: Alwyn Eades {jae5-at-lehigh.edu} 10, 22 -- Organization: Lehigh University 10, 22 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 10, 22 -- X-Accept-Language: en-us, en 10, 22 -- MIME-Version: 1.0 10, 22 -- To: "MicroscopyListserver (E-mail)" {Microscopy-at-microscopy.com} 10, 22 -- Subject: Free TEM Workshop Materials and Geology 10, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 22 -- Content-Transfer-Encoding: 7bit 10, 22 -- X-Virus-Scanned: ClamAV version 0.88, clamav-milter version 0.87 on rain.CC.Lehigh.EDU 10, 22 -- X-Virus-Status: Clean ==============================End of - Headers==============================
The Quanta 200 uses a W filament. Unless it is a SFEG. If there are no ion pumps, you should be able to use Nitrogen or He. If there is an ion pump, do not use He. It will kill the pump.
I found that for VP (20-120Pa) that N2 works fine and is better than air (less vacuum issues). So, for ESEM, I would think that N2 ought to be the gas of choice as well.
For VP mode, I do quant with EDAX Genesis using collected values at two pressure/vacuum values. This is their VIP option. I find good correlation between it and high vacuum mode. Which EDS are you using? Does it have an ESEM option mode?
gary g.
At 06:45 AM 2/1/2006, you wrote:
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==============================Original Headers============================== 12, 20 -- From gary-at-gaugler.com Wed Feb 1 20:27:16 2006 12, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k122RF4w004457 12, 20 -- for {microscopy-at-microscopy.com} ; Wed, 1 Feb 2006 20:27:16 -0600 12, 20 -- Received: (qmail 10416 invoked from network); 1 Feb 2006 18:27:15 -0800 12, 20 -- Received: by simscan 1.1.0 ppid: 10403, pid: 10405, t: 0.1857s 12, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 20 -- by qsmtp2 with SMTP; 1 Feb 2006 18:27:14 -0800 12, 20 -- Message-Id: {6.2.3.4.2.20060201181844.0204ac20-at-mail.calweb.com} 12, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 12, 20 -- Date: Wed, 01 Feb 2006 18:27:16 -0800 12, 20 -- To: hagglundk1-at-nku.edu 12, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 20 -- Subject: Re: [Microscopy] ESEM chamber gas choices 12, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 20 -- In-Reply-To: {200602011445.k11EjI9l000396-at-ns.microscopy.com} 12, 20 -- References: {200602011445.k11EjI9l000396-at-ns.microscopy.com} 12, 20 -- Mime-Version: 1.0 12, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
it is a long time ago I have used a Tyrode-GA-mixture as a perfusion fixative for rat brain (selective hypothalamic target preparation containing ventricle III as well as magnocellular nuclear regions, providing "open" blood vessels/capillaries in that region; followed by a 3-dim reconstruction of those nuclear regions)....
Later on I had to use also such perfusion mixtures for retrograde perfusion of pig (renal) arteries...... I don't know or have at hand now the original receipt of } Tyrode {'s Mammalian Ringer solution, but have found in my methods/techniques collection } old { descriptions and protocols of the respective solutions and measurement data.
I have seen that your mixing formula varies only in the amount of Glucose......you indicate: 1 gm, in my descriptions I do have listed 10 grms.
On the other hand, when working with Tyrode's, initially I haven't mixed the buffer with sodium cacodylate, but instead with a very small amount of sodium-dihydrogenphosphate as follows: NaCl: 6.0 gm KCl: 0.2 gm CaCl2 (as CaCl2.2H2O) 0.2 (0.25g) gm MgCl2 (as MgCl2.6H2O) 0.1 (0.2. g) gm sodium-dihydrogenphosphate 0.05 gm Sodium-hydrogencarbonate(NaHCO3) 1.0 gm Glucose 10.0 gm ------------------------------------------------------------------------ ---- ad 1000 ml A.bidest (DD A.dest.) Phosphate(s) must be dissolved extra and should finally be added when all other } powder-substances { (! especially CaCl2 and MgCl2) are readily dissolved.
I have noted in the protocol: initial pH of the resulting solution: 8.4, approx. 275-277 mosmol (measuring also pH 8.14, mosmol: 310, 290, 285, 282, 295, 295).
pH-correction with approx. 2.95 ml 1 N HCl to pH. 7.2
The fixative for perfusion consisted of: 160 ml 25% glutaraldehyde (which means a final GA-concentration of 4% in the fixative) to be mixed with the above mentioned Tyrode stock solution to an endvolume of 1000 ml. If there formed a pale/ milky precipitate (which sometimes occured, perhaps due to a poor GA-quality with a high amount of aldehyde polymers - we had at that time in the late 1970ies - but most probably due to the Na2HPO4/NaH2PO4 !) the solution should be / has to be filtrated.
After doing so, I noted: pH ==} 8.4, 545-550 mosmol
Since the pH of that fix-mixture increased (????) to 8.4 with time again, it was necessary to adjust again with some ml of 1 N HCl to a pH of at least 7.7 (the solution now seemed to act quite well as a } buffer { system ! ) and finally, having adjusted the solution to pH 7.2 again, I then measured 1160 /1190 / 1200 mosmol.
As the following washing buffer I used at that time a Tyrode solution as given above, but added Glucose 25 g ad 1000 ml solution (which perhaps was wrong), and measured (after pH correction with approx. 1.3 ml of 1 N HCl to pH 7.2) 360 / 370 / 375 mosmol.
The 4% glutaraldehyde perhaps will contribute approx. 280 mosmol to the total osmolarity (if calculating from some own measurements and some tables found in the literaturefor 1, 1.5, 2.0 and 5 % GA-Solutions, respectively), the rest up to 1200 mosmol therefore must originate from the (more complete) dissociation of total ions dissolved in the tyrode's solution.
You state that the Tyrode solution you measured had 400/401 mosmol and is thought to be unusually high.
I assume that there is a contribution by the sodium-cacodylate you added (which is, if you use 10.7 g / 1000 ml, a final molarity of 0.05 mol) which could be in the range of 100 mosmol.
Consider also that due to more effective dissociation of ionic components in the solution the more diluted your fluid is, osmolarity will increase - not in a 1:1 mode (eg. Na-phosphate ions in Na2HPO4 and NaH2PO4 0,1 M will not have double the osmol amount of 0.05 M, which in fact will be higher due to more effective dissociation).
In my files I have other/additional data concerning the use of Tyrode's ringer solution, I you like, I could share those with you on request offline.
Best regards,
Wolfgang Muss OR Dr. Wolfgang Muss EM-Lab ====} with pride: 25 years in operation by 2nd of Feb. 2006 {==== Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
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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 dyel-at-mail.nih.gov as well as the MIcroscopy Listserver --------------------------------------------------------------------------- Email: dyel-at-mail.nih.gov Name: Chip Dye Organization: NIH-NICHD Title-Subject: [Filtered] Tyrodes-Cacodylate Buffer
Question: Dear ListServer:
Has anyone used this buffer before? I plan on using this to make up a fixative to perfuse fix a rat. I will be looking at myelin of the sciatic nerve. Why does this buffer have such a high osmolarity? Any thoughts?
There are some things that are not clear about this problem. The question was written as if the use of water in an ESEM is a problem specifically for silicon drift detectors. Surely the problem is the same for all types of detector; the problem is not with the detector but with the window. In a system working correctly, the vacuum of the detector is quite separate from the vacuum of the chamber. So it does not matter what gas you use in the ESEM as long as the window is intact.
Does water in an ESEM cause the window of the detector to develop holes more quickly than, say, nitrogen in the ESEM? This could be the case, depending on the window design. We did have window failures in our ESEM, but we are told that this was a temporary problem and that the windows being sold now are just fine in an ESEM using water. Does anyone have specific knowledge on this?
Alwyn Eades
-- ........... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
==============================Original Headers============================== 6, 25 -- From jae5-at-lehigh.edu Thu Feb 2 09:36:46 2006 6, 25 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k12FakDr007269 6, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Feb 2006 09:36:46 -0600 6, 25 -- Received: from [128.180.55.141] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 6, 25 -- (authenticated bits=0) 6, 25 -- by rain.CC.Lehigh.EDU (8.13.5/8.13.5) with ESMTP id k12Fakid004491 6, 25 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 6, 25 -- Thu, 2 Feb 2006 10:36:46 -0500 6, 25 -- Message-ID: {43E2270E.4050600-at-lehigh.edu} 6, 25 -- Date: Thu, 02 Feb 2006 10:36:46 -0500 6, 25 -- From: Alwyn Eades {jae5-at-lehigh.edu} 6, 25 -- Organization: Lehigh University 6, 25 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 6, 25 -- X-Accept-Language: en-us, en 6, 25 -- MIME-Version: 1.0 6, 25 -- To: hagglundk1-at-nku.edu, 6, 25 -- "MicroscopyListserver (E-mail)" {Microscopy-at-microscopy.com} 6, 25 -- Subject: Re: [Microscopy] ESEM chamber gas choices 6, 25 -- References: {200602011533.k11FXYuO006193-at-ns.microscopy.com} 6, 25 -- In-Reply-To: {200602011533.k11FXYuO006193-at-ns.microscopy.com} 6, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 25 -- Content-Transfer-Encoding: 7bit 6, 25 -- X-Virus-Scanned: ClamAV version 0.88, clamav-milter version 0.87 on rain.CC.Lehigh.EDU 6, 25 -- X-Virus-Status: Clean ==============================End of - Headers==============================
Does anyone have experience with the Cytoviva imaging system. It looks to me (from their website at www.cytovita.com like it is a retrofit condenser. They claim a resolution better than 50 nm. In fact, their website says "The typical optical microscope provides a maximum theoretical resolution limit of 240nm. With resolution below 100nm and detection below 50nm, CytoViva allows you to see details never before possible with traditional microscopy. " Can someone explain to me how one can improve on the maximum theoretical resolution?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Dear Prof Phillips, perhaps the instrument of Cytoviva ( http://www.cytoviva.com ) is a further development of a technique I found in PNAS 2002 (if you want I can provide you with a pdf of this article). Unfortunately I have not followed that thread.....
FYI: Title of work:
Fast 100-nm resolution three-dimensional microscope reveals structural plasticity of mitochondria in live yeast Alexander Egner, Stefan Jakobs, and Stefan W. Hell*
3370-3375 PNAS March 19, 2002 vol. 99 no. 6 By introducing beam-scanning multifocal multiphoton 4Pi-confocal microscopy, we have attained fast fluorescence imaging of live cells with axial super resolution. Rapid scanning of up to 64 pairs of interfering high-angle fields and subsequent confocal detection enabled us to perform three to five times finer optical sectioning than confocal microscopy. In conjunction with nonlinear image restoration, we demonstrate, to our knowledge for the first time, three-dimensional imaging of live eukaryotic cells at an equilateral resolution of ~ 100 nm. This imaging mode allowed us to reveal the morphology and size of the green fluorescent protein-labeled mitochondrial compartment of live Saccharomyces cerevisiae (bakers' yeast) growing on different carbon sources. Our studies show that mitochondria of cells grown on medium containing glycerol as the only carbon source, as opposed to glucose-grown cells, exhibit a strongly branched tubular reticulum. We determine the average tubular diameter and find that it increases from 339 +/- 5 nm to 360 +/- 4 nm when changing from glucose to glycerol, that is, from a fermentable to a nonfermentable carbon source. Moreover, this change is associated with a 2.8-fold increase of the surface of the reticulum, resulting in an average increase in volume of the mitochondrial compartment by a factor of 3.0 +/- 0.2. Best regards
Wolfgang Muss Salzburg
---------- Von: phillipst-at-missouri.edu[SMTP:phillipst-at-missouri.edu] Antwort an: phillipst-at-missouri.edu Gesendet: Donnerstag, 02. Februar 2006 18:44 An: W.Muss-at-salk.at Betreff: [Microscopy] Cytoviva system
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Does anyone have experience with the Cytoviva imaging system. It looks to me (from their website at www.cytovita.com like it is a retrofit condenser. They claim a resolution better than 50 nm. In fact, their website says "The typical optical microscope provides a maximum theoretical resolution limit of 240nm. With resolution below 100nm and detection below 50nm, CytoViva allows you to see details never before possible with traditional microscopy. " Can someone explain to me how one can improve on the maximum theoretical resolution?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
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Hi Alwyn,
My information on this is not definite but, my understanding is that different EDS manufacturers use different process for producing their detector windows. Consequently, there are different consequences between detectors depending on the gases used in the chamber. I can't see that there would be a difference between Si(Li) and SDD but I can understand a difference between manufacturers.
Best regards, -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
==============================Original Headers============================== 5, 16 -- From larry-at-cymru.freewire.co.uk Thu Feb 2 14:51:22 2006 5, 16 -- Received: from get.freewire.net (get.freewire.net [195.184.229.250]) 5, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k12KpLst006559 5, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 2 Feb 2006 14:51:22 -0600 5, 16 -- Received: from [217.154.254.216] ([217.154.254.216]) 5, 16 -- by get.freewire.net (8.11.6/8.11.6) with ESMTP id k12KpH004925; 5, 16 -- Thu, 2 Feb 2006 20:51:17 GMT 5, 16 -- Mime-Version: 1.0 5, 16 -- Message-Id: {p06210200c0081f8a361d-at-[217.154.254.125]} 5, 16 -- In-Reply-To: {200602021537.k12FbpsN009683-at-ns.microscopy.com} 5, 16 -- References: {200602021537.k12FbpsN009683-at-ns.microscopy.com} 5, 16 -- Date: Thu, 2 Feb 2006 20:48:41 +0000 5, 16 -- To: jae5-at-lehigh.edu, Microscopy-at-MSA.Microscopy.Com 5, 16 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 16 -- Subject: [Microscopy] Re: ESEM chamber gas choices 5, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
} My information on this is not definite but, my understanding is that } different EDS manufacturers use different process for producing their } detector windows. Consequently, there are different consequences } between detectors depending on the gases used in the chamber. I can't } see that there would be a difference between Si(Li) and SDD but I can } understand a difference between manufacturers.
To bring this back to SDD, my understanding is that all SDD detector chips are manufactured by KETEK ... While individual SDD detector manufacturers will choose to employ different windows. I am still waiting to find out the source of information regarding a general query about SDDs' incompatibility with water vapor.
Genuinely, Michael Shaffer :o)
SEM/MLA Laboratory Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 7, 21 -- From michael-at-shaffer.net Thu Feb 2 16:31:55 2006 7, 21 -- Received: from ws6-5.us4.outblaze.com (ws6-5.us4.outblaze.com [205.158.62.152]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k12MVtbM017649 7, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Feb 2006 16:31:55 -0600 7, 21 -- Received: (qmail 9845 invoked from network); 2 Feb 2006 22:31:55 -0000 7, 21 -- Received: from unknown (HELO rarewolf1) (michael-at-shaffer.net-at-205.251.84.119) 7, 21 -- by ws6-5.us4.outblaze.com with SMTP; 2 Feb 2006 22:31:54 -0000 7, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 21 -- To: {larry-at-cymru.freewire.co.uk} , 7, 21 -- "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 21 -- Subject: RE: [Microscopy] Re: ESEM chamber gas choices 7, 21 -- Date: Thu, 2 Feb 2006 19:01:39 -0330 7, 21 -- Message-ID: {000201c62848$706e48f0$4f01a8c0-at-rarewolf1} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 7, 21 -- Thread-Index: AcYoOqzcXnO24mvMRVios5iM13rJLgADMGFA 7, 21 -- In-Reply-To: {200602022052.k12KqIrL007363-at-ns.microscopy.com} ==============================End of - Headers==============================
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Question: For those who do lab service for hire, how much is the going rate for SEM/EDX work? I only do work within our corporation but we're putting together a cost of what our work would cost if it was done outside.
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Email: mmstuason-at-yahoo.ca Name: Lizette Tuason
Title-Subject: [Filtered] Type of CO2 for CPD
Question: Hi everyone,
What type of CO2 do you use for CPD? The CO2 we currently have hooked up to our Denton DCP-1 is supercritical fluid extraction grade but it costs more than CAD$750. I looked up some online sites where people mentioned that they use just standard CO2 (in the US). Iím not sure what standard CO2 is and what the equivalent would be here. Iím buying from Praxair (Canada) and there are several categories that Iím not sure which one I should be choosing.
Could anyone whoís doing CPD tell me what CO2 you buy ń company and catalog number?
$200/hour....$150/hour.... $125/hour...$100/hour....pick a number.
This is a nice set of numbers but there is more to outsourcing than just the hourly rate...IMO. If a specimen is prepared in-house and then sent out-house, what is the out-house person to look for? Well, you will have to spend x hours writing a detailed procedural instruction for what you want to be analyzed. If the specimen deviates from this, then what? What is your writing time worth? Nothing?
The point is that there is great value in having the requester sitting by the SEM operator. Look here, look there, what is this, what is that? If you outsource, you will get exactly or less than what you ask for since the operator does not know what to do outside of your written request. If the job is so mundane (how many are?) then this is not a problem. The SEM is a very valuable tool for many areas of interest. But given a 12mm diameter specimen stub, which 2u are of the most interest? "I'll know it when I see it." Right. but they threw the specimen over the wall to the SEM folks. The results thrown back may not match up with what was needed. The variability of specimens means that there is no simple approach to evaluation. Plus, the requester is not at the SEM to know that they saw what they needed the first time.
gary g.
Disclaimer: I do not do this sort of ambiguous work. However, I do perform SEM analysis of unknown specimens and known specimens but I know what to look for.
At 05:44 PM 2/2/2006, you wrote:
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You are partially right about CytoViva: it is a retrofittable condenser fitted with special optics and an illuminator system.
Re: Resolution: I worked with Aetos at some length on the launch of CytoViva and have seen better than 90nm resolution (as measured with a Richardson test slide).
Re: how it works Dr. Vitaly Vodyanoy, the inventor currently has a paper in progress which will explain the physics. My personal observations lead me to believe that there is some sort of resonance effect rather than the traditional scattering or diffraction we normally use for imaging. The result looks a lot like fluorescence (bright object/dark background), without the need for staining. It also has an interesting ability to optically section (much like DIC or confocal). We were able to watch spirochetes actually invading cells as well as see more detail in bacteria and some special marine cells.
I encourage you to visit their website, if only to see some interesting images. Their technical applications specialist, Dr. Tom Hasling, is usually quite happy to run samples and Byron Cheatham, their sales manager, can probably arrange for a demo in your lab, if you are serious about potential purchase.
The system compares extremely well to other systems on the market in the $150K range, at about 10% the cost. ... and there's no cost for looking!
Hope this was helpful, Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
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 11:42 AM 2/2/2006, phillipst-at-missouri.edu wrote:
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==============================Original Headers============================== 22, 17 -- From bfoster-at-mme1.com Fri Feb 3 03:05:42 2006 22, 17 -- Received: from smtp109.sbc.mail.mud.yahoo.com (smtp109.sbc.mail.mud.yahoo.com [68.142.198.208]) 22, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1395gKj030224 22, 17 -- for {microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 03:05:42 -0600 22, 17 -- Received: (qmail 98384 invoked from network); 3 Feb 2006 09:05:41 -0000 22, 17 -- Received: from unknown (HELO barbsd505.mme1.com) (bfostermme-at-sbcglobal.net-at-68.94.37.39 with login) 22, 17 -- by smtp109.sbc.mail.mud.yahoo.com with SMTP; 3 Feb 2006 09:05:41 -0000 22, 17 -- Message-Id: {7.0.1.0.0.20060203025348.0204e158-at-mme1.com} 22, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 22, 17 -- Date: Fri, 03 Feb 2006 03:05:43 -0600 22, 17 -- To: phillipst-at-missouri.edu, Microscopy Listserver {microscopy-at-microscopy.com} 22, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 22, 17 -- Subject: Re: [Microscopy] Cytoviva system 22, 17 -- In-Reply-To: {200602021742.k12HggDM022539-at-ns.microscopy.com} 22, 17 -- References: {200602021742.k12HggDM022539-at-ns.microscopy.com} 22, 17 -- Mime-Version: 1.0 22, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
I'd recommend you also look into the Hi-Scope from Hirox-USA. They provide some really interesting imaging modes and have been well accepted by a number of major customers here in the US.
Caveat: MME has no financial interest in this product.
Hope this 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
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 08:32 PM 1/30/2006, brad.huggins-at-bp.com wrote:
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==============================Original Headers============================== 15, 17 -- From bfoster-at-mme1.com Fri Feb 3 03:15:06 2006 15, 17 -- Received: from smtp107.sbc.mail.mud.yahoo.com (smtp107.sbc.mail.mud.yahoo.com [68.142.198.206]) 15, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k139F6LN007226 15, 17 -- for {microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 03:15:06 -0600 15, 17 -- Received: (qmail 40367 invoked from network); 3 Feb 2006 09:15:06 -0000 15, 17 -- Received: from unknown (HELO barbsd505.mme1.com) (bfostermme-at-sbcglobal.net-at-68.94.37.39 with login) 15, 17 -- by smtp107.sbc.mail.mud.yahoo.com with SMTP; 3 Feb 2006 09:15:06 -0000 15, 17 -- Message-Id: {7.0.1.0.0.20060201092440.01ff2348-at-mme1.com} 15, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 17 -- Date: Fri, 03 Feb 2006 03:15:08 -0600 15, 17 -- To: brad.huggins-at-bp.com, Microscopy Listserver {microscopy-at-microscopy.com} 15, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 15, 17 -- Subject: Re: [Microscopy] viaWWW: LM, Anyone with "Digital Microscope" 15, 17 -- In-Reply-To: {200601310232.k0V2WUBj013521-at-ns.microscopy.com} 15, 17 -- References: {200601310232.k0V2WUBj013521-at-ns.microscopy.com} 15, 17 -- Mime-Version: 1.0 15, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
You don't need the really pure stuff. We use food-grade, and in one trial found it cleaner (less water, oil, and particulates) than the expensive, "pure" siphon CO2. Just be sure to use a siphon-CO2 (comes in a cylinder with a siphon tube, so you get liquid, not gas, coming out), and spend the money for filters and dehydrating sieves.
Phil
We have the same Denton you do and use something what Airgas calls "bone dry", cat. No. CDBD200S with siphon tube. In addition, we use the Tousimis liquid CO2 filter (cat. No. 8784, I believe it helps). We pay about US$ 65 for a cylinder, if I remember well. This seems to be giving good results for CPD of biological material (mostly mouse embryos) for SEM.
Hope this helps,
Michal
mmstuason-at-yahoo.ca wrote:
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==============================Original Headers============================== 7, 21 -- From M_Jarnik-at-fccc.edu Fri Feb 3 08:51:46 2006 7, 21 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k13EpjM8030588 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 08:51:46 -0600 7, 21 -- Received: from fccc.edu (emf4.fccc.edu [131.249.9.170]) 7, 21 -- by azah.fccc.edu (8.12.11/8.12.11) with ESMTP id k13Epj1t020965 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 09:51:45 -0500 (EST) 7, 21 -- Message-ID: {43E36E02.2040108-at-fccc.edu} 7, 21 -- Date: Fri, 03 Feb 2006 09:51:46 -0500 7, 21 -- From: Michal Jarnik {M_Jarnik-at-fccc.edu} 7, 21 -- Reply-To: M_Jarnik-at-fccc.edu 7, 21 -- Organization: Fox Chase Cancer Center 7, 21 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 7, 21 -- X-Accept-Language: en,cs 7, 21 -- MIME-Version: 1.0 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- Subject: Re: [Microscopy] viaWWW: type of CO2 do you use for CPD 7, 21 -- References: {200602030142.k131gpqb032061-at-ns.microscopy.com} 7, 21 -- In-Reply-To: {200602030142.k131gpqb032061-at-ns.microscopy.com} 7, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 21 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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Email: Jones_e1269-at-yahoo.com Name: Jason Jones
Organization: The University of Texas Health Science Center at San Antonio
Title-Subject: [Filtered] Immuno EM
Question:
I am looking for a facility to do immuno-EM on the yeast Candida albicans for us. We're interested in intracellular localization of a soluble secretory protein, secreted aspartyl protease (Sap2p) in wild-type and a secretory mutant strain we have generated.
We have polyclonal antibodies to Sap2p, which have worked quite well in the published literature for immunoEM, and we have also 6X-His tagged this protein in our wild-type and mutant strains.
IWe do not have immunoEM available as a core facility here.
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Email: df-at-donnforbes.com Name: Donn Forbes
Organization: Arasil, Inc.
Title-Subject: [Filtered] SEM for BWA Detection
Question: Why isn't SEM a standard technology for detecting and identifying viruses and bacteria used as biological warfare agents?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bcarrington-at-slfc.org) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, February 3, 2006 at 09:15:17 ---------------------------------------------------------------------------
Email: bcarrington-at-slfc.org Name: Brenda Carrington
Organization: The Learning Center
Education: K-8 Grade Grammar School
Location: Chesterfeild MO (St. Louis area)
Question: We are haing a microscope day on Feb. 27 from 10:00-3:00. I don't know how to contact a local person to assist us. We have over 100 kids grades 2-9. We will be studying microbes and microscopes and using the GEMS materials.
We have inherited lots of copper TEM grids that are decades old. Many of them have a dark patina on them, and, on those grids, we have been losing our sections during alcohol-based uranyl acetate staining. The sections remain when we use water-based uranyl acetate, however. We have tried sonicating the grids in ethanol, acetone, or chloroform, none of which has solved our problem.
Does anyone have any suggestions for cleaning them so that we will not lose our sections during staining, or should we pitch them? We really do prefer to stain with alcohol- based uranyl acetate, since it provides a more intense staining.
As always, thanks for any suggestions you might have.
Dotty Sorenson
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006 7, 17 -- Received: from pushingtin.mr.itd.umich.edu (pushingtin.mr.itd.umich.edu [141.211.14.78]) 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k13LmwV1020156 7, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Feb 2006 15:48:58 -0600 7, 17 -- Received: from [141.214.170.47] (host-47.subnet-170.med.umich.edu [141.214.170.47]) 7, 17 -- by pushingtin.mr.itd.umich.edu (smtp) with ESMTP id k13LmvJZ004383; 7, 17 -- Fri, 3 Feb 2006 16:48:57 -0500 7, 17 -- Mime-Version: 1.0 (Apple Message framework v746.2) 7, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 17 -- Message-Id: {F847072A-3C48-4995-B594-5900360673CB-at-umich.edu} 7, 17 -- Cc: Sasha Meshinchi {meshin-at-umich.edu} 7, 17 -- Content-Transfer-Encoding: 7bit 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 7, 17 -- Subject: Cleaning TEM grids 7, 17 -- Date: Fri, 3 Feb 2006 16:47:15 -0500 7, 17 -- To: microscopy-at-msa.microscopy.com 7, 17 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
Hi Dotty, I would try putting your grids into a drying oven for at least 5 minutes after you cut your ultrathin sections. After they are totally dried down, you can take them out. If you forget and leave your grids in the oven overnight, it doesn't seem to hurt anything at all. The heat seems to bond the sections to the grids very well, such that I've never had an experience of sections coming off the grids, regardless of the state of the grids.
To clean my grids, I just dip them into concentrated sodium hydroxide for a few seconds, and then rinse them by dipping them a few times in distilled water just before I pick up the sections. It sort of etches the grids.
Garry Burgess Charge Technologist - Electron Microscopy Health Science Centre Winnipeg, Canada
Dear listers,
We have inherited lots of copper TEM grids that are decades old. Many of them have a dark patina on them, and, on those grids, we have been losing our sections during alcohol-based uranyl acetate staining. The sections remain when we use water-based uranyl acetate, however. We have tried sonicating the grids in ethanol, acetone, or chloroform, none of which has solved our problem.
Does anyone have any suggestions for cleaning them so that we will not lose our sections during staining, or should we pitch them? We really do prefer to stain with alcohol- based uranyl acetate, since it provides a more intense staining.
As always, thanks for any suggestions you might have.
Dotty Sorenson
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006 7, 17 -- Received: from pushingtin.mr.itd.umich.edu (pushingtin.mr.itd.umich.edu [141.211.14.78]) 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k13LmwV1020156 7, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Feb 2006 15:48:58 -0600 7, 17 -- Received: from [141.214.170.47] (host-47.subnet-170.med.umich.edu [141.214.170.47]) 7, 17 -- by pushingtin.mr.itd.umich.edu (smtp) with ESMTP id k13LmvJZ004383; 7, 17 -- Fri, 3 Feb 2006 16:48:57 -0500 7, 17 -- Mime-Version: 1.0 (Apple Message framework v746.2) 7, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 17 -- Message-Id: {F847072A-3C48-4995-B594-5900360673CB-at-umich.edu} 7, 17 -- Cc: Sasha Meshinchi {meshin-at-umich.edu} 7, 17 -- Content-Transfer-Encoding: 7bit 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 7, 17 -- Subject: Cleaning TEM grids 7, 17 -- Date: Fri, 3 Feb 2006 16:47:15 -0500 7, 17 -- To: microscopy-at-msa.microscopy.com 7, 17 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
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==============================Original Headers============================== 14, 20 -- From GBurgess-at-exchange.hsc.mb.ca Fri Feb 3 16:08:11 2006 14, 20 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 14, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k13M8A8U017896 14, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 16:08:11 -0600 14, 20 -- Received: from hscxntmx0007.hsc.mb.ca (unverified [172.16.6.179]) by 14, 20 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 14, 20 -- {B0017955251-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Fri, 14, 20 -- 3 Feb 2006 16:08:04 -0600 14, 20 -- Received: by hscxntmx0007.hsc.mb.ca with Internet Mail Service 14, 20 -- (5.5.2653.19)id {DXXMZXB1} ; Fri, 3 Feb 2006 16:08:43 -0600 14, 20 -- Message-ID: 14, 20 -- {00A937989100304A83A058F6C45873FF32A393-at-hscxntmx0005.hsc.mb.ca} 14, 20 -- Date: Fri, 3 Feb 2006 16:04:40 -0600 14, 20 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 14, 20 -- Subject: RE: [Microscopy] Cleaning TEM grids 14, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 14, 20 -- X-Mailer: Internet Mail Service (5.5.2653.19) 14, 20 -- MIME-Version: 1.0 14, 20 -- Content-Type: text/plain; 14, 20 -- charset="iso-8859-1" ==============================End of - Headers==============================
We clean our grids in acid alcohol, a mixture of 3% HCL in 95% ethanol. Cover the grids in a small shell vial with the acid alcohol, swirl intermittantly for 30 to 40 seconds or so and rinse well with 95% ethanol. If the grids have a very dark patina then clean for a longer time. You'll know when they are done because they'll have a shiny, bright copper finish. We usually clean many grids at a time, but you could clean them one by one by dipping the grid in the acid alcohol and then rinsing in a stream of 95% ETOH just before you use them. This works better on grids that have been cleaned, but have set around for a while, long enough to have developed a small oxidized layer. Dark patinas like the grids you have will probably need to be put into a vial and cleaned as described above.
dsoren-at-umich.edu wrote:
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-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
==============================Original Headers============================== 6, 21 -- From gstrout-at-ou.edu Fri Feb 3 17:33:43 2006 6, 21 -- Received: from c3p0.ou.edu (mail.zero.ou.edu [129.15.0.75]) 6, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k13NXhBV028012 6, 21 -- for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 3 Feb 2006 17:33:43 -0600 6, 21 -- Received: from [127.0.0.1] ([129.15.38.202]) 6, 21 -- by c3p0.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 6, 21 -- 2005)) with ESMTP id {0IU400EHDXG5Q5A0-at-c3p0.ou.edu} for 6, 21 -- Microscopy-at-Sparc5.Microscopy.Com; Fri, 03 Feb 2006 17:33:41 -0600 (CST) 6, 21 -- Date: Fri, 03 Feb 2006 17:33:40 -0600 6, 21 -- From: Greg Strout {gstrout-at-ou.edu} 6, 21 -- Subject: Re: [Microscopy] Cleaning TEM grids 6, 21 -- In-reply-to: {200602032150.k13LoAHd025474-at-ns.microscopy.com} 6, 21 -- To: Microscopy List Server {Microscopy-at-ns.Microscopy.Com} , dsoren-at-umich.edu 6, 21 -- Message-id: {43E3E854.7040107-at-ou.edu} 6, 21 -- MIME-version: 1.0 6, 21 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 21 -- Content-transfer-encoding: 7BIT 6, 21 -- X-Accept-Language: en-us, en 6, 21 -- References: {200602032150.k13LoAHd025474-at-ns.microscopy.com} 6, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 21 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
Try soaking a few grids in store grade ammonia for about 1/2 hr to 1 hr to see if the patina is removed at all. Rinse well in distilled water and see what happens.
-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: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu] Sent: Friday, February 03, 2006 1:53 PM To: Walck-at-SouthBayTech.com
Dear listers,
We have inherited lots of copper TEM grids that are decades old. Many of them have a dark patina on them, and, on those grids, we have been losing our sections during alcohol-based uranyl acetate staining. The sections remain when we use water-based uranyl acetate, however. We have tried sonicating the grids in ethanol, acetone, or chloroform, none of which has solved our problem.
Does anyone have any suggestions for cleaning them so that we will not lose our sections during staining, or should we pitch them? We really do prefer to stain with alcohol- based uranyl acetate, since it provides a more intense staining.
As always, thanks for any suggestions you might have.
Dotty Sorenson
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006 7, 17 -- Received: from pushingtin.mr.itd.umich.edu (pushingtin.mr.itd.umich.edu [141.211.14.78]) 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k13LmwV1020156 7, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Feb 2006 15:48:58 -0600 7, 17 -- Received: from [141.214.170.47] (host-47.subnet-170.med.umich.edu [141.214.170.47]) 7, 17 -- by pushingtin.mr.itd.umich.edu (smtp) with ESMTP id k13LmvJZ004383; 7, 17 -- Fri, 3 Feb 2006 16:48:57 -0500 7, 17 -- Mime-Version: 1.0 (Apple Message framework v746.2) 7, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 17 -- Message-Id: {F847072A-3C48-4995-B594-5900360673CB-at-umich.edu} 7, 17 -- Cc: Sasha Meshinchi {meshin-at-umich.edu} 7, 17 -- Content-Transfer-Encoding: 7bit 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 7, 17 -- Subject: Cleaning TEM grids 7, 17 -- Date: Fri, 3 Feb 2006 16:47:15 -0500 7, 17 -- To: microscopy-at-msa.microscopy.com 7, 17 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
==============================Original Headers============================== 17, 27 -- From walck-at-southbaytech.com Fri Feb 3 20:16:08 2006 17, 27 -- Received: from ylpvm15.prodigy.net (ylpvm15-ext.prodigy.net [207.115.57.46]) 17, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k142G7Bg007081 17, 27 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 20:16:08 -0600 17, 27 -- Received: from pimout7-ext.prodigy.net (pimout7-int.prodigy.net [207.115.4.147]) 17, 27 -- by ylpvm15.prodigy.net (8.12.10 outbound/8.12.10) with ESMTP id k142GKEV029956 17, 27 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Feb 2006 21:16:20 -0500 17, 27 -- X-ORBL: [64.169.193.90] 17, 27 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 17, 27 -- by pimout7-ext.prodigy.net (8.13.4 outbound domainkey aix/8.13.4) with ESMTP id k142Fvmf095186; 17, 27 -- Fri, 3 Feb 2006 21:16:02 -0500 17, 27 -- From: "Scott Walck" {walck-at-southbaytech.com} 17, 27 -- To: {dsoren-at-umich.edu} 17, 27 -- Cc: {Microscopy-at-microscopy.com} 17, 27 -- Subject: RE: [Microscopy] Cleaning TEM grids 17, 27 -- Date: Fri, 3 Feb 2006 18:16:01 -0800 17, 27 -- Message-ID: {007801c62930$f81347e0$7801a8c0-at-dynamicbl8uno3} 17, 27 -- MIME-Version: 1.0 17, 27 -- Content-Type: text/plain; 17, 27 -- charset="us-ascii" 17, 27 -- Content-Transfer-Encoding: 7bit 17, 27 -- X-Priority: 3 (Normal) 17, 27 -- X-MSMail-Priority: Normal 17, 27 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 17, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 17, 27 -- In-Reply-To: {200602032153.k13LrLc5007111-at-ns.microscopy.com} 17, 27 -- Importance: Normal ==============================End of - Headers==============================
I've always cleaned grids by quickly passing through the flame of an alcohol burner. Quick and easy. Works on my old grids, though they aren't decades old!
Diana van Driel Dept Ophthalmology Sydney University GPO Box 4337 Sydney, NSW AUSTRALIA 2001
Phone 61 2 93827278 Mobile 0423 151614 FAX 61 2 93827318 On 4 Feb 2006, at 8:50 AM, dsoren-at-umich.edu wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Dear listers, } } We have inherited lots of copper TEM grids that are decades old. } Many of them have a dark patina on them, and, on those grids, we have } been losing our sections during alcohol-based uranyl acetate } staining. The sections remain when we use water-based uranyl } acetate, however. We have tried sonicating the grids in ethanol, } acetone, or chloroform, none of which has solved our problem. } } Does anyone have any suggestions for cleaning them so that we will } not lose our sections during staining, or should we pitch them? We } really do prefer to stain with alcohol- based uranyl acetate, since } it provides a more intense staining. } } As always, thanks for any suggestions you might have. } } Dotty Sorenson } } Dorothy Sorenson } Microscopy and Image-analysis Laboratory } Department of Cell and Developmental Biology } University Of Michigan Medical School } 4643 Medical Science Building II } 1301 Catherine } Ann Arbor, MI 48109-0616 } (734)763-1170 } FAX (734)763-1166 } } } ==============================Original } Headers============================== } 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006 } 7, 17 -- Received: from pushingtin.mr.itd.umich.edu } (pushingtin.mr.itd.umich.edu [141.211.14.78]) } 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k13LmwV1020156 } 7, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Feb 2006 } 15:48:58 -0600 } 7, 17 -- Received: from [141.214.170.47] } (host-47.subnet-170.med.umich.edu [141.214.170.47]) } 7, 17 -- by pushingtin.mr.itd.umich.edu (smtp) with ESMTP id } k13LmvJZ004383; } 7, 17 -- Fri, 3 Feb 2006 16:48:57 -0500 } 7, 17 -- Mime-Version: 1.0 (Apple Message framework v746.2) } 7, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } format=flowed } 7, 17 -- Message-Id: {F847072A-3C48-4995-B594-5900360673CB-at-umich.edu} } 7, 17 -- Cc: Sasha Meshinchi {meshin-at-umich.edu} } 7, 17 -- Content-Transfer-Encoding: 7bit } 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu} } 7, 17 -- Subject: Cleaning TEM grids } 7, 17 -- Date: Fri, 3 Feb 2006 16:47:15 -0500 } 7, 17 -- To: microscopy-at-msa.microscopy.com } 7, 17 -- X-Mailer: Apple Mail (2.746.2) } ==============================End of - } Headers============================== }
==============================Original Headers============================== 6, 20 -- From dianavd-at-eye.usyd.edu.au Sun Feb 5 22:06:54 2006 6, 20 -- Received: from galen.med.usyd.edu.au (machaon.med.usyd.edu.au [129.78.36.30]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1646r3u022150 6, 20 -- for {Microscopy-at-microscopy.com} ; Sun, 5 Feb 2006 22:06:53 -0600 6, 20 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) 6, 20 -- by galen.med.usyd.edu.au with esmtp (Exim 4.50) 6, 20 -- id 1F5xYY-0004NY-9a 6, 20 -- for Microscopy-at-microscopy.com; Mon, 06 Feb 2006 15:00:52 +1100 6, 20 -- Mime-Version: 1.0 (Apple Message framework v623) 6, 20 -- In-Reply-To: {200602032150.k13Loevb027732-at-ns.microscopy.com} 6, 20 -- References: {200602032150.k13Loevb027732-at-ns.microscopy.com} 6, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 20 -- Message-Id: {25a1f04e5a26d06c447521476709da97-at-eye.usyd.edu.au} 6, 20 -- Content-Transfer-Encoding: 7bit 6, 20 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} 6, 20 -- Subject: Re: [Microscopy] Cleaning TEM grids 6, 20 -- Date: Mon, 6 Feb 2006 15:06:32 +1100 6, 20 -- To: Microscopy {Microscopy-at-microscopy.com} 6, 20 -- X-Mailer: Apple Mail (2.623) 6, 20 -- X-Spam-Score: -5.9 (-----) ==============================End of - Headers==============================
Dear Dotty, We were in the same situation some years ago. We cleaned the grids in the following way: - put the grids into small beaker (25ml) filled with 5% - 10% hydrochloric acid - let the grids to clean for some minutes, gently shake several times (at the end of cleaning the grids should be shiny gold) - remove the cleaning hydrochloric acid solution and wash the grids several times with distilled water - remove the distilled water as much as possible from the beaker and replace it with acetone and let stand for some time - pour the acetone with the grids into clean glass Petri dish filled with filter paper - remove the filter paper with the grids and put it into another glass Petri dish and let dry out the rest of acetone - that's all
The troubles with losing the sections during the staining could be overcome by making the grids sticky: - put 10 ml of chloroform into 25 ml glass beaker - cut about 5 cm of 3M Magic Scotch tape and wash it in the chloroform - remove the rest of the tape from the beaker - put the grids onto filter paper and drop the "sticky solution" on them using Pasteur pipette - let the grids dry and use them for collecting the sections
Best regards from Prague Oldrich ---------------------------------------- Oldrich Benada Institute of Microbiology Acad. Sci. CR Videnska 1083 CZ - 142 20 Prague 4 Czech Republic ---------------------------------------
} } } ---------------------------------------------------------------------------- } 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, } } We have inherited lots of copper TEM grids that are decades old. } Many of them have a dark patina on them, and, on those grids, we have } been losing our sections during alcohol-based uranyl acetate } staining. The sections remain when we use water-based uranyl } acetate, however. We have tried sonicating the grids in ethanol, } acetone, or chloroform, none of which has solved our problem. } } Does anyone have any suggestions for cleaning them so that we will } not lose our sections during staining, or should we pitch them? We } really do prefer to stain with alcohol- based uranyl acetate, since } it provides a more intense staining. } } As always, thanks for any suggestions you might have. } } Dotty Sorenson } } Dorothy Sorenson } Microscopy and Image-analysis Laboratory } Department of Cell and Developmental Biology } University Of Michigan Medical School } 4643 Medical Science Building II } 1301 Catherine } Ann Arbor, MI 48109-0616 } (734)763-1170 } FAX (734)763-1166 } } } ==============================Original } Headers============================== } 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006 } 7, 17 -- Received: from pushingtin.mr.itd.umich.edu } (pushingtin.mr.itd.umich.edu [141.211.14.78]) } 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k13LmwV1020156 } 7, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Feb 2006 15:48:58 } -0600 } 7, 17 -- Received: from [141.214.170.47] (host-47.subnet-170.med.umich.edu } [141.214.170.47]) } 7, 17 -- by pushingtin.mr.itd.umich.edu (smtp) with ESMTP id } k13LmvJZ004383; } 7, 17 -- Fri, 3 Feb 2006 16:48:57 -0500 } 7, 17 -- Mime-Version: 1.0 (Apple Message framework v746.2) } 7, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } format=flowed } 7, 17 -- Message-Id: {F847072A-3C48-4995-B594-5900360673CB-at-umich.edu} } 7, 17 -- Cc: Sasha Meshinchi {meshin-at-umich.edu} } 7, 17 -- Content-Transfer-Encoding: 7bit } 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu} } 7, 17 -- Subject: Cleaning TEM grids } 7, 17 -- Date: Fri, 3 Feb 2006 16:47:15 -0500 } 7, 17 -- To: microscopy-at-msa.microscopy.com } 7, 17 -- X-Mailer: Apple Mail (2.746.2) } ==============================End of - } Headers============================== }
==============================Original Headers============================== 9, 25 -- From benada-at-biomed.cas.cz Mon Feb 6 03:33:26 2006 9, 25 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 9, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k169XPV1003261 9, 25 -- for {microscopy-at-microscopy.com} ; Mon, 6 Feb 2006 03:33:26 -0600 9, 25 -- Received: from mail2.biomed.cas.cz (localhost [127.0.0.1]) 9, 25 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k169V0cA006059; 9, 25 -- Mon, 6 Feb 2006 10:31:00 +0100 (CET) 9, 25 -- Received: from 147.231.44.104 9, 25 -- (SquirrelMail authenticated user benada) 9, 25 -- by mail2.biomed.cas.cz with HTTP; 9, 25 -- Mon, 6 Feb 2006 10:31:01 +0100 (CET) 9, 25 -- Message-ID: {1280.147.231.44.104.1139218261.squirrel-at-mail2.biomed.cas.cz} 9, 25 -- In-Reply-To: {200602032152.k13LqKBS002784-at-ns.microscopy.com} 9, 25 -- References: {200602032152.k13LqKBS002784-at-ns.microscopy.com} 9, 25 -- Date: Mon, 6 Feb 2006 10:31:01 +0100 (CET) 9, 25 -- Subject: Re: [Microscopy] Cleaning TEM grids 9, 25 -- From: =?iso-8859-2?Q?Old=F8ich_Benada?= {benada-at-biomed.cas.cz} 9, 25 -- To: dsoren-at-umich.edu 9, 25 -- Cc: microscopy-at-microscopy.com 9, 25 -- User-Agent: SquirrelMail/1.4.4 9, 25 -- MIME-Version: 1.0 9, 25 -- Content-Type: text/plain;charset=iso-8859-2 9, 25 -- Content-Transfer-Encoding: 8bit 9, 25 -- X-Priority: 3 (Normal) 9, 25 -- Importance: Normal ==============================End of - Headers==============================
Overview: Schafer Corporation, Sunol, California, is seeking a skilled and innovative individual to add to our mass spectrometry group. SVL performs materials characterization and related analytical services on commercial and government contracts. The activities of the group include chemical and elemental analysis of materials on a production basis, maintenance of several mass spectrometers and ancillary equipment, development of new or improved MS analysis techniques, and quality assurance of analytical data.
Schafer is a technically strong firm with a reputation for quality and integrity. Schafer's reputation is a direct result of our dedicated, motivated, talented, and creative staff that is responsible for developing our outstanding business relationships with our customers. Our technical capabilities are vast and growing to provide innovations for the future.
The successful candidate will work as part of a team responsible for processing and analyzing small samples using laboratory instrumentation, primarily Thermal Ionization and/or Secondary Ion Mass Spectrometers.
Responsibilities: Duties include instrument operation and data interpretation; filament construction and preparation; standards loading; as well as high vacuum, high voltage, and cryogenic system maintenance. The successful candidate should have or be able to develop skills to process samples, maintain equipment, and assist in developing and optimizing analytical techniques.
Qualifications: The ideal candidate will have technical training in an appropriate physical sciences field and related experience in a scientific laboratory, preferably operating mass spectrometers or other analytical equipment. Ability to work independently as well as part of a team is required. Good customer service focus and commitment to quality are required. Experience in the use of a light microscope, and sufficient physical dexterity to perform micromanipulation tasks is highly desired, as is knowledge of high vacuum and cryogenic principles. Schafer is looking for people that have established safe laboratory skills and exceptional aptitude for details including accurate record keeping. Experience in laboratory data management (word processing, spread sheets and databases) is also desired. Experience with mechanical systems or machining skills would be helpful. Other qualifications include: . AA degree plus 10 years experience or Bachelor's degree in physical science or engineering plus two years of technical experience. . Demonstrated ability to solve technical problems. . Experience in data evaluation and quality control. . Must be a US citizen with the ability to obtain government security clearance. Apply at: http://www.schafercorp.com/Careers/open.htm http://jobs-schafer.icims.com/schafer_jobs/jobs/candidate/job.jsp?jobid=1106 &mode=view
==============================Original Headers============================== 6, 20 -- From dsams-at-schaferlabs.com Mon Feb 6 19:16:53 2006 6, 20 -- Received: from mail.schaferlabs.com (mail.schaferlabs.com [64.168.91.154]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k171GkxV006745 6, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Feb 2006 19:16:52 -0600 6, 20 -- Received: from dsamsws ([10.10.10.1]) 6, 20 -- by mail.schaferlabs.com (8.12.11/8.12.11) with ESMTP id k171ANrk020002 6, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Feb 2006 17:10:34 -0800 6, 20 -- Message-Id: {200602070110.k171ANrk020002-at-mail.schaferlabs.com} 6, 20 -- From: "David Sams" {dsams-at-schaferlabs.com} 6, 20 -- To: {Microscopy-at-microscopy.com} 6, 20 -- Subject: Mass Spectrometry Scientist / Senior Engineer at Schafer Corporation 6, 20 -- Date: Mon, 6 Feb 2006 17:16:18 -0800 6, 20 -- MIME-Version: 1.0 6, 20 -- Content-Type: text/plain; 6, 20 -- charset="US-ASCII" 6, 20 -- Content-Transfer-Encoding: 7bit 6, 20 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 6, 20 -- In-reply-to: 6, 20 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 6, 20 -- Thread-Index: AcVrdaWs9qR/q27tSgm/q8Axr3R/miwGADdQAAH4L3AAM6jdwAPH8fGQ ==============================End of - Headers==============================
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Email: RANGETS-at-AOL.COM Name: BEN GHAFFAARI
Organization: RANGE TECHNICAL
Title-Subject: [Filtered] NEED SCHEMATICS
Question: DOES ANYONE HAVE AN EXTRA SET OF "LEO" 440 SCHEMATICS, WE CAN BUY OR PURCHASE, WHEN WE TRY TO CONTACT THE MFR, WE DO NOT GET A RESPONSE, SO MAYBE WE ARE USING AN INCORRECT CONTACT. IF SOMEONE KNOWS WHO WE CONTACT FOR "LEO" PARTS ANY SUGGESTIONS WILL BE APPRECIATED
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 mcmahojt-at-ccf.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mcmahojt-at-ccf.org Name: Jim McMahon
Organization: Cleveland Clinic Foundation
Title-Subject: [Filtered] Spurr Resin
Question: We have been having great difficulty with the newly formulated Spurr Resin and in particular the ERL component. Not only is Spurr no longer low viscosity but we find that its penetration and polymerization to be far inferior to the original. We are prepared to abandon the resin in favor of another such as PolyBed or Araldite. But first I would like to know if anyone else has had similar problems and was able to solve them.
for what it is worth, i've recently re-tried a commercial version of Epon 812 on a cell suspension. i found the cells would not settle through the resin mix, and could not be pelleted. a parallel embedment in the new Spurr with ERL behaved very well. in short, whatever is said about low viscosity editions of Epon, neither they, nor the original were ever thin enough to allow embedments of free cells and i personally found them more than a little difficult to cut. but then i've always found Epon, in whatever formulation, prone to static and difficult to section.
on the other hand, the problem may really be an issue of density of the medium, causing the the cells to be bouyant in the medium. does anyone want to contribute knowledge on that?
having said that, i do find the new ERL component to be a little brittle, but very sectionable.
paul
==============================Original Headers============================== 7, 21 -- From paul_hazelton-at-umanitoba.ca Mon Feb 6 23:49:49 2006 7, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k175niDC005523 7, 21 -- for {microscopy-at-microscopy.com} ; Mon, 6 Feb 2006 23:49:48 -0600 7, 21 -- Received: from [130.179.152.145] (cvx-082.cc.umanitoba.ca [130.179.152.145]) 7, 21 -- (authenticated bits=0) 7, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k175ndFk026699; 7, 21 -- Mon, 6 Feb 2006 23:49:40 -0600 (CST) 7, 21 -- Message-ID: {43E9865F.3010305-at-umanitoba.ca} 7, 21 -- Date: Tue, 07 Feb 2006 23:49:19 -0600 7, 21 -- From: Paul Hazelton {paul_hazelton-at-umanitoba.ca} 7, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 7, 21 -- X-Accept-Language: en-us, en 7, 21 -- MIME-Version: 1.0 7, 21 -- To: mcmahojt-at-ccf.org 7, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 7, 21 -- Subject: Re: [Microscopy] viaWWW: Spurr Resin 7, 21 -- References: {200602070432.k174Wx2a027920-at-ns.microscopy.com} 7, 21 -- In-Reply-To: {200602070432.k174Wx2a027920-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==============================
We would need some to improve cell attachment for a time course experiment. Surprisingly, we were not able to locate any coverslips (unlike slides). Does anybody have recommendations? (Of course, we can always make our own.)
Thanks,
Michael
==============================Original Headers============================== 6, 19 -- From M_Jarnik-at-fccc.edu Tue Feb 7 08:35:31 2006 6, 19 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 6, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k17EZVJJ022275 6, 19 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Feb 2006 08:35:31 -0600 6, 19 -- Received: from fccc.edu (emf4.fccc.edu [131.249.9.170]) 6, 19 -- by azah.fccc.edu (8.12.11/8.12.11) with ESMTP id k17EZU5d016976 6, 19 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Feb 2006 09:35:31 -0500 (EST) 6, 19 -- Message-ID: {43E8B032.7060002-at-fccc.edu} 6, 19 -- Date: Tue, 07 Feb 2006 09:35:30 -0500 6, 19 -- From: Michal Jarnik {M_Jarnik-at-fccc.edu} 6, 19 -- Reply-To: M_Jarnik-at-fccc.edu 6, 19 -- Organization: Fox Chase Cancer Center 6, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 6, 19 -- X-Accept-Language: en,cs 6, 19 -- MIME-Version: 1.0 6, 19 -- To: microscopy-at-msa.microscopy.com 6, 19 -- Subject: Poly-L-lysine coated coverslips 6, 19 -- Content-Type: text/plain; charset=us-ascii; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I have recently encountered problems with Spurr as well. I embed 100 micron thick Vibratome sections of brain so penetration should not be a problem. I am using the same processing proceedure I have used for many, many years on much larger tissue blocks. The center of the sections seems to be poorly infiltrated and the block is very brittle. Cutting intact 1 micron sections is nearly impossible. The viscosity seems 'normal', in other words, like I am used to with Spurr and I am not familiar with any 'new' or 'improved' version of the mix or its components. I will cut some more blocks and talk to my supplier (a very reputable EM supplier I have used for 30 years) to see if I can shed some light on this problem.
Geoff
mcmahojt-at-ccf.org wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 8, 32 -- From mcauliff-at-umdnj.edu Tue Feb 7 08:43:50 2006 8, 32 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 8, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k17Ehnhp031730 8, 32 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Feb 2006 08:43:49 -0600 8, 32 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 8, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 9D1FD4BE3B 8, 32 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Feb 2006 08:43:49 -0600 (CST) 8, 32 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 8, 32 -- by zix02.umdnj.edu (Proprietary) with ESMTP id 59FA44BE34 8, 32 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Feb 2006 08:43:48 -0600 (CST) 8, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 32 -- id {0IUB00D01MP310-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 32 -- for microscopy-at-msa.microscopy.com; Tue, 07 Feb 2006 09:43:44 -0500 (EST) 8, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 8, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 32 -- 2004)) with ESMTP id {0IUB00BK4NAVAK-at-Polaris.umdnj.edu} ; Tue, 8, 32 -- 07 Feb 2006 09:37:43 -0500 (EST) 8, 32 -- Date: Tue, 07 Feb 2006 09:37:06 -0500 8, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 32 -- Subject: Re: [Microscopy] Spurr Resin 8, 32 -- In-reply-to: {200602070432.k174W3M3025365-at-ns.microscopy.com} 8, 32 -- To: mcmahojt-at-ccf.org, MicroscopyListserver {microscopy-at-msa.microscopy.com} , 8, 32 -- paul_hazelton-at-umanitoba.ca 8, 32 -- Message-id: {43E8B092.8030903-at-umdnj.edu} 8, 32 -- MIME-version: 1.0 8, 32 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 8, 32 -- Content-transfer-encoding: 7BIT 8, 32 -- X-Accept-Language: en-us, en 8, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 32 -- Gecko/20040804 Netscape/7.2 (ax) 8, 32 -- References: {200602070432.k174W3M3025365-at-ns.microscopy.com} ==============================End of - Headers==============================
Depends on what you want to do. If it is for EM you can use the Thermonox coverslips. I think all EM suppliers sell them, along with the general suppliers.
However, if you want to do IF or confocal work the thermonox will quench the reactions somewhat so you will have to do glass. I've used straight glass, but many cell lines will not stick well to glass. Undoubtably someone will provide the name of a supplier of pre-treated slips if they are out there somewhere.
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 Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
==============================Original Headers============================== 9, 21 -- From paul_hazelton-at-umanitoba.ca Tue Feb 7 08:53:50 2006 9, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 9, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k17ErnB5008760 9, 21 -- for {microscopy-at-microscopy.com} ; Tue, 7 Feb 2006 08:53:49 -0600 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 k17ErhaH011897; 9, 21 -- Tue, 7 Feb 2006 08:53:44 -0600 (CST) 9, 21 -- Message-ID: {43E8B474.8040005-at-umanitoba.ca} 9, 21 -- Date: Tue, 07 Feb 2006 08:53:40 -0600 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: M_Jarnik-at-fccc.edu 9, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 9, 21 -- Subject: Re: [Microscopy] Poly-L-lysine coated coverslips 9, 21 -- References: {200602071438.k17EcWLj026119-at-ns.microscopy.com} 9, 21 -- In-Reply-To: {200602071438.k17EcWLj026119-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==============================
Regarding the brittle nature of Spurrs resin due to the new ERL component, we have solved our problems by combining the components according to the "soft" recipe:
ERL 10g DER 7g NSA 26g DMAE 0.4ml
This mixture results in blocks about as hard as the old "hard" formulation using VCD. We have not noticed a lack of tissue penetration during infiltration, but we may be less sensitive to this issue. Polymerization is still at 70 deg C for 17 hours.
Best wishes,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
-----Original Message----- X-from: mcmahojt-at-ccf.org [mailto:mcmahojt-at-ccf.org] Sent: Monday, February 06, 2006 8:38 PM To: drk-at-SHCC.org
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Email: mcmahojt-at-ccf.org Name: Jim McMahon
Organization: Cleveland Clinic Foundation
Title-Subject: [Filtered] Spurr Resin
Question: We have been having great difficulty with the newly formulated Spurr Resin and in particular the ERL component. Not only is Spurr no longer low viscosity but we find that its penetration and polymerization to be far inferior to the original. We are prepared to abandon the resin in favor of another such as PolyBed or Araldite. But first I would like to know if anyone else has had similar problems and was able to solve them.
I think that they are responding in their own way. Their answer is "No."
gary g.
At 08:30 PM 2/6/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Tue Feb 7 10:06:45 2006 9, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k17G6jtp028532 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 7 Feb 2006 10:06:45 -0600 9, 20 -- Received: (qmail 12166 invoked from network); 7 Feb 2006 08:06:43 -0800 9, 20 -- Received: by simscan 1.1.0 ppid: 12163, pid: 12164, t: 0.1441s 9, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 20 -- by qsmtp1 with SMTP; 7 Feb 2006 08:06:43 -0800 9, 20 -- Message-Id: {6.2.3.4.2.20060207080513.02057d00-at-mail.calweb.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 9, 20 -- Date: Tue, 07 Feb 2006 08:06:43 -0800 9, 20 -- To: RANGETS-at-AOL.COM 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] viaWWW: NEED SCHEMATICS LEO 440 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200602070430.k174UXNn021696-at-ns.microscopy.com} 9, 20 -- References: {200602070430.k174UXNn021696-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
what you recommend in your note is the logical solution. actually, i had meant to try the same change for my last embedment of monolayers to correct for the hardness/brittleness. unfortunately, i got distracted and forgot. i suspect that to get the medium hardness you may need 8-9 ml of the ERL component.
also, i used to use 0.4gm DMAE but changed that to .3gm to give a longer pot life. also, higher catalyst concentration could have an effect on viscosity and penetration.
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 Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
==============================Original Headers============================== 10, 21 -- From paul_hazelton-at-umanitoba.ca Tue Feb 7 15:19:25 2006 10, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 10, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k17LJPwp019538 10, 21 -- for {microscopy-at-microscopy.com} ; Tue, 7 Feb 2006 15:19:25 -0600 10, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 10, 21 -- (authenticated bits=0) 10, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k17LJFkD028260; 10, 21 -- Tue, 7 Feb 2006 15:19:24 -0600 (CST) 10, 21 -- Message-ID: {43E90ED0.7030901-at-umanitoba.ca} 10, 21 -- Date: Tue, 07 Feb 2006 15:19:12 -0600 10, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 10, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 10, 21 -- X-Accept-Language: en-us, en 10, 21 -- MIME-Version: 1.0 10, 21 -- To: DRK-at-SHCC.org 10, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 10, 21 -- Subject: Re: [Microscopy] RE: viaWWW: Spurr Resin 10, 21 -- References: {200602071558.k17Fw6YT022905-at-ns.microscopy.com} 10, 21 -- In-Reply-To: {200602071558.k17Fw6YT022905-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==============================
An immediate opening exists for a skilled microscopist in Dow's Global Analytical Sciences Laboratory part of Dow's Corporate R&D function. We are seeking an individual with a passion for characterization science and a desire to both apply this science to complex industrial problems and to advance the technology. A Ph.D. in science is preferred, but an individual with a Masters degree and relevant work experience will also be considered. Working experience in electron microscopy is essential with skills in scanning and transmission electron microscopy, focused ion beam methods, energy dispersive x-ray analysis, electron energy loss spectroscopy, electron diffraction and microtomy preferred. A background in catalysis is a plus. Excellent written and oral communication skills (English) are essential with an ability to work in a globally diverse team environment. The position supports new product development activities working from a laboratory with a FEG TEM-STEM, 2 TEMs, a FIBSEM, EPMA, FEGSEM with cryotransfer system, a full complement of scanned probe and light microscopes, SAXS, XPS and TOF-SIMS. The position is located in Midland, Michigan. Applicants must have the ability to work in the USA.
Dow is a leader in science and technology, providing innovative chemical, plastic and agricultural products and services to many essential consumer markets. With annual sales of $40 billion, Dow serves customers in 175 countries and a wide range of markets that are vital to human progress: food, transportation, health and medicine, personal and home care, and building and construction, among others. Committed to the principles of sustainable development, Dow and its 43,000 employees seek to balance economic, environmental and social responsibilities. References to "Dow" or the "Company" mean The Dow Chemical Company and its consolidated subsidiaries unless otherwise expressly noted.
Please submit curriculum vitae and a list of references to Dr. John Blackson, Building 1897, The Dow Chemical Company, Midland, MI 48667.
Best Regards, Bill William A. Heeschen, Ph.D. Microscopy, Digital Imaging The Dow Chemical Company Midland, MI 48674 waheeschen-at-dow.com
==============================Original Headers============================== 7, 23 -- From WAHeeschen-at-dow.com Tue Feb 7 16:05:53 2006 7, 23 -- Received: from mail86.messagelabs.com (mail86.messagelabs.com [216.82.244.115]) 7, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k17M5q3V029468 7, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 7 Feb 2006 16:05:52 -0600 7, 23 -- X-VirusChecked: Checked 7, 23 -- X-Env-Sender: WAHeeschen-at-dow.com 7, 23 -- X-Msg-Ref: server-4.tower-86.messagelabs.com!1139349950!29545829!1 7, 23 -- X-StarScan-Version: 5.5.9.1; banners=-,-,- 7, 23 -- X-Originating-IP: [204.136.184.23] 7, 23 -- Received: (qmail 26574 invoked from network); 7 Feb 2006 22:05:51 -0000 7, 23 -- Received: from mail6.dow.com (HELO txnte41.nam.dow.com) (204.136.184.23) 7, 23 -- by server-4.tower-86.messagelabs.com with SMTP; 7 Feb 2006 22:05:51 -0000 7, 23 -- Received: by TXNTE41.nam.dow.com with Internet Mail Service (5.5.2658.3) 7, 23 -- id {1A02KCZD} ; Tue, 7 Feb 2006 16:05:50 -0600 7, 23 -- Message-ID: {CC55EF96AD3142438EF779F71571702084A20F-at-USMDLMDOWX004.dow.com} 7, 23 -- From: "Heeschen, Bill (WA)" {WAHeeschen-at-dow.com} 7, 23 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 7, 23 -- Subject: Job Posting from The Dow Chemical Company 7, 23 -- Date: Tue, 7 Feb 2006 16:05:31 -0600 7, 23 -- Expiry-Date: Tue, 6 Feb 2007 23:00:00 -0600 7, 23 -- MIME-Version: 1.0 7, 23 -- X-Mailer: Internet Mail Service (5.5.2658.3) 7, 23 -- Content-Type: text/plain ==============================End of - Headers==============================
I just got off the phone with Stacy (the owner) at Electron Microscopy Sciences. I discussed my recent problems with Spurr's resin with her. She told me that one component, ERL 4206, is no longer maunfactured due to its high toxicity. It has been replaced with ERL 4221. She thinks that this is the cause of recent problems with Spurr embedments. She has discussed this with others (I did not ask who) and the consensus recommendations are to prolong dehydration in graded ethanols to 20 minutes each step, make the 1:1 prop. oxide:Spurr step 4 hours and make the first change of pure resin 6 hours or longer. Since longer times in solvents are known to extract cytoplasmic components (and I cannot imaging that a 100 micron Vibratome section of CNS needs 20 min. per change of graded ethanol) I am going back to Epon substitutes. Also, my tissues are in a second change of fresh resin on a rotator overnight and then spend at least 1 hour under vacuum, I can't see how infiltration could be a problem. I have used both the soft and firm mixtures with different tissues and had inconsistant block textures and infiltration problems with both. I don't have these problems with an Araldite kit I bought at the same time. I can't risk knock-out mice in long-term treatment/recovery experiments to reagents I cannot be sure of.
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, 29 -- From mcauliff-at-umdnj.edu Wed Feb 8 15:25:23 2006 6, 29 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 6, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k18LPNGG000454 6, 29 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Feb 2006 15:25:23 -0600 6, 29 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 6, 29 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id BC9CB4BE54 6, 29 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Feb 2006 16:25:22 -0500 (EST) 6, 29 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 6, 29 -- by zix03.umdnj.edu (Proprietary) with ESMTP id 063404BE5C 6, 29 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Feb 2006 16:25:22 -0500 (EST) 6, 29 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 29 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 29 -- id {0IUD00E01ZO0X6-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 29 -- for microscopy-at-msa.microscopy.com; Wed, 08 Feb 2006 16:25:21 -0500 (EST) 6, 29 -- Received: from [127.0.0.1] ([10.138.2.240]) 6, 29 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 29 -- 2004)) with ESMTP id {0IUD00DAGZSNRH-at-Polaris.umdnj.edu} for 6, 29 -- microscopy-at-msa.microscopy.com; Wed, 08 Feb 2006 16:02:47 -0500 (EST) 6, 29 -- Date: Wed, 08 Feb 2006 16:02:08 -0500 6, 29 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 29 -- Subject: Spurr's resin update 6, 29 -- To: MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 29 -- Message-id: {43EA5C50.6070607-at-umdnj.edu} 6, 29 -- MIME-version: 1.0 6, 29 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 29 -- Content-transfer-encoding: 7BIT 6, 29 -- X-Accept-Language: en-us, en 6, 29 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 29 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
I will be experimenting with alternative scintillators for our Hitachi S4800 STEM unit. As I was advised to no touch the scintillator in the microscope, I hope someone will be able to offer measurements for the original S4800 scintillator.
Thanks, Daniel Salamon Technical Officer, Electron Microscopy
National Institute for Nanotechnology, NRC W6-017A ECERF Bldg, 9107-116 Street Edmonton, AB. T6G 2V4
We are just starting an experiment with en-bloc staining of tissue in uranyl acetate in 70% ethanol. It seems to me that since the blocks are larger than thin sections, they probably take up more radioactive material- so should we take any precautions when handling the blocks after the resin has cured? (i.e. store the blocks in separate special containers, wear gloves when sectioning or are there any special cleaning procedures for the knife?).
thank you for sharing your knowledge
Gerd
Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
I am seeking to buy NanoScope AFM equipment, such as MultiMode or Dimension, to use with my Nanoscope IIIA controller. If you have anything to sell, please contact me offline.
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
==============================Original Headers============================== 3, 21 -- From donc-at-asmicro.com Thu Feb 9 06:56:12 2006 3, 21 -- Received: from smtp112.sbc.mail.re2.yahoo.com (smtp112.sbc.mail.re2.yahoo.com [68.142.229.93]) 3, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k19CuBAg016578 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 06:56:11 -0600 3, 21 -- Received: (qmail 34978 invoked from network); 9 Feb 2006 12:56:11 -0000 3, 21 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.57.205.80 with login) 3, 21 -- by smtp112.sbc.mail.re2.yahoo.com with SMTP; 9 Feb 2006 12:56:11 -0000 3, 21 -- Message-ID: {006801c62d77$ebd28b80$c901a8c0-at-ASM11} 3, 21 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- To: "Microscopy List" {microscopy-at-microscopy.com} 3, 21 -- Subject: AFM equipment wanted 3, 21 -- Date: Thu, 9 Feb 2006 07:54:03 -0500 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: text/plain; 3, 21 -- charset="iso-8859-1" 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Priority: 3 3, 21 -- X-MSMail-Priority: Normal 3, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 3, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
We are looking to purchase a second-hand Gatan 622SC camera (200kV-compatible) in decent working condition to install on our JEOL 2100 TEM. We are hoping to find one with a working intensifier and an intact scintillator, although any problems are possibly acceptable. We would be able to pay for it using a University of Maryland PO. The amount of money might allow you to upgrade your TEM to a digital CCD camera from AMT.
Please do not reply to the listserver. Direct any inquiries or offers directly to me at the email address below.
Many thanks,
-John
-- John Cumings cumings-at-umd.edu Assistant Professor Department of Materials Science and Engineering University of Maryland College Park, MD 20742-2115
office (301) 405-0789 (1246 Kim Building) fax (301) 314-8164 --
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We have a few users study bacterial pili using negative staining with PTA (pH 6.5). We have got some pretty nice images. But often time, we could not find pili even on the positive control. When we did see pili, they were not on all bacteria in the same sample. Is this a common phenomenon? Do bacteria lose their pili easily when the external condition is not favorable? if that is the case, what should be done to minimize the loss.
Thank you in advance.
Hong Emory EM
==============================Original Headers============================== 5, 22 -- From hyi-at-emory.edu Thu Feb 9 10:06:53 2006 5, 22 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19G6qLb005145 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 10:06:53 -0600 5, 22 -- Received: from [170.140.232.204] (localhost [127.0.0.1]) 5, 22 -- by virginia.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k19G6qIx024009 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 11:06:52 -0500 (EST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v622) 5, 22 -- Message-Id: {56be70af163d5629d409dc9e17804801-at-emory.edu} 5, 22 -- Content-Type: text/plain; 5, 22 -- charset=ISO-8859-1; 5, 22 -- format=flowed 5, 22 -- To: Microscopy-at-microscopy.com 5, 22 -- From: Hong Yi {hyi-at-emory.edu} 5, 22 -- Subject: Bacterial Pili 5, 22 -- Date: Thu, 9 Feb 2006 11:06:51 -0500 5, 22 -- X-Mailer: Apple Mail (2.622) 5, 22 -- X-imss-version: 2.037 5, 22 -- X-imss-result: Passed 5, 22 -- X-imss-approveListMatch: *-at-emory.edu 5, 22 -- Content-Transfer-Encoding: 8bit 5, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19G6qLb005145 ==============================End of - Headers==============================
We often have this same problem. At least in our lab, flagella and pili seem to hide when we're trying to find them and show up when we don't care if we see them or not.
As a rule, they seem to be easily detached by rough handling, including centrifuging, and maybe by the staining process itself, since we often find them isolated from the bacteria on the grid.
One group of researchers gets around this problem by growing the bacteria on carbon coated grids and staining them in situ. Check JOURNAL OF BACTERIOLOGY, Feb. 2005, p. 1173-1181 for images. For preparation details this article references Brown et al, 2001. Immunocytochemical localization of HrpA and HrpZ supports a role for the Hrp pilus in the transfer of effector proteins from Pseudomonassyringae pv. tomato across the host plant cell wall. Mol. Plant-Microbe Interact. 14:394-404. I have not yet read the last article, so I can't comment on it.
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: hyi-at-emory.edu [mailto:hyi-at-emory.edu] Sent: Thursday, February 09, 2006 10:08 AM To: Tindall, Randy D.
Dear Microscopists:
We have a few users study bacterial pili using negative staining with PTA (pH 6.5). We have got some pretty nice images. But often time, we could not find pili even on the positive control. When we did see pili, they were not on all bacteria in the same sample. Is this a common phenomenon? Do bacteria lose their pili easily when the external condition is not favorable? if that is the case, what should be done to minimize the loss.
Thank you in advance.
Hong Emory EM
==============================Original Headers============================== 5, 22 -- From hyi-at-emory.edu Thu Feb 9 10:06:53 2006 5, 22 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19G6qLb005145 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 10:06:53 -0600 5, 22 -- Received: from [170.140.232.204] (localhost [127.0.0.1]) 5, 22 -- by virginia.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k19G6qIx024009 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 11:06:52 -0500 (EST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v622) 5, 22 -- Message-Id: {56be70af163d5629d409dc9e17804801-at-emory.edu} 5, 22 -- Content-Type: text/plain; 5, 22 -- charset=ISO-8859-1; 5, 22 -- format=flowed 5, 22 -- To: Microscopy-at-microscopy.com 5, 22 -- From: Hong Yi {hyi-at-emory.edu} 5, 22 -- Subject: Bacterial Pili 5, 22 -- Date: Thu, 9 Feb 2006 11:06:51 -0500 5, 22 -- X-Mailer: Apple Mail (2.622) 5, 22 -- X-imss-version: 2.037 5, 22 -- X-imss-result: Passed 5, 22 -- X-imss-approveListMatch: *-at-emory.edu 5, 22 -- Content-Transfer-Encoding: 8bit 5, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19G6qLb005145 ==============================End of - Headers==============================
==============================Original Headers============================== 20, 24 -- From TindallR-at-missouri.edu Thu Feb 9 10:24:23 2006 20, 24 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 20, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19GOMnC014637 20, 24 -- for {microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 10:24:22 -0600 20, 24 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 20, 24 -- Thu, 9 Feb 2006 10:24:22 -0600 20, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 20, 24 -- Content-class: urn:content-classes:message 20, 24 -- MIME-Version: 1.0 20, 24 -- Content-Type: text/plain; 20, 24 -- charset="iso-8859-1" 20, 24 -- Subject: RE: [Microscopy] Bacterial Pili 20, 24 -- Date: Thu, 9 Feb 2006 10:24:21 -0600 20, 24 -- Message-ID: {BA876152E8653240BE8572E897083EE701849D33-at-UM-EMAIL09.um.umsystem.edu} 20, 24 -- X-MS-Has-Attach: 20, 24 -- X-MS-TNEF-Correlator: 20, 24 -- Thread-Topic: [Microscopy] Bacterial Pili 20, 24 -- thread-index: AcYtkwuBClshBOD6QsCkENhoqgFICQAAJ20Q 20, 24 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 20, 24 -- To: {hyi-at-emory.edu} 20, 24 -- Cc: {microscopy-at-microscopy.com} 20, 24 -- X-OriginalArrivalTime: 09 Feb 2006 16:24:22.0502 (UTC) FILETIME=[49041860:01C62D95] 20, 24 -- Content-Transfer-Encoding: 8bit 20, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19GOMnC014637 ==============================End of - Headers==============================
Rough handling may contribute to loss of pili, however the PTA itself is probably the culprit. Try changing the pH either up or down. Way back when, I did a study with rotavirus and various negative stains. Bottom line is that the length of time of staining, pH and concentration all contributed to the destruction (preservation) of the viral particles. If I remember correctly, the length of time was the most important factor. Also try using either uranyl acetate or ammonium molybdate as the negative stain.
Best of luck,
Ed
Edward P. Calomeni Director EM Lab - Pathology The Ohio State University M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210 614-293-5580 (office) 614-293-8806 (lab) edward.calomeni-at-osumc.edu -----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Thursday, February 09, 2006 11:29 AM To: Edward Calomeni
Hong,
We often have this same problem. At least in our lab, flagella and pili seem to hide when we're trying to find them and show up when we don't care if we see them or not.
As a rule, they seem to be easily detached by rough handling, including centrifuging, and maybe by the staining process itself, since we often find them isolated from the bacteria on the grid.
One group of researchers gets around this problem by growing the bacteria on carbon coated grids and staining them in situ. Check JOURNAL OF BACTERIOLOGY, Feb. 2005, p. 1173-1181 for images. For preparation details this article references Brown et al, 2001. Immunocytochemical localization of HrpA and HrpZ supports a role for the Hrp pilus in the transfer of effector proteins from Pseudomonassyringae pv. tomato across the host plant cell wall. Mol. Plant-Microbe Interact. 14:394-404. I have not yet read the last article, so I can't comment on it.
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: hyi-at-emory.edu [mailto:hyi-at-emory.edu] Sent: Thursday, February 09, 2006 10:08 AM To: Tindall, Randy D.
Dear Microscopists:
We have a few users study bacterial pili using negative staining with PTA (pH 6.5). We have got some pretty nice images. But often time, we could not find pili even on the positive control. When we did see pili, they were not on all bacteria in the same sample. Is this a common phenomenon? Do bacteria lose their pili easily when the external condition is not favorable? if that is the case, what should be done to minimize the loss.
Thank you in advance.
Hong Emory EM
==============================Original Headers============================== 5, 22 -- From hyi-at-emory.edu Thu Feb 9 10:06:53 2006 5, 22 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19G6qLb005145 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 10:06:53 -0600 5, 22 -- Received: from [170.140.232.204] (localhost [127.0.0.1]) 5, 22 -- by virginia.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k19G6qIx024009 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 11:06:52 -0500 (EST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v622) 5, 22 -- Message-Id: {56be70af163d5629d409dc9e17804801-at-emory.edu} 5, 22 -- Content-Type: text/plain; 5, 22 -- charset=ISO-8859-1; 5, 22 -- format=flowed 5, 22 -- To: Microscopy-at-microscopy.com 5, 22 -- From: Hong Yi {hyi-at-emory.edu} 5, 22 -- Subject: Bacterial Pili 5, 22 -- Date: Thu, 9 Feb 2006 11:06:51 -0500 5, 22 -- X-Mailer: Apple Mail (2.622) 5, 22 -- X-imss-version: 2.037 5, 22 -- X-imss-result: Passed 5, 22 -- X-imss-approveListMatch: *-at-emory.edu 5, 22 -- Content-Transfer-Encoding: 8bit 5, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19G6qLb005145 ==============================End of - Headers==============================
==============================Original Headers============================== 20, 24 -- From TindallR-at-missouri.edu Thu Feb 9 10:24:23 2006 20, 24 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 20, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19GOMnC014637 20, 24 -- for {microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 10:24:22 -0600 20, 24 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 20, 24 -- Thu, 9 Feb 2006 10:24:22 -0600 20, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 20, 24 -- Content-class: urn:content-classes:message 20, 24 -- MIME-Version: 1.0 20, 24 -- Content-Type: text/plain; 20, 24 -- charset="iso-8859-1" 20, 24 -- Subject: RE: [Microscopy] Bacterial Pili 20, 24 -- Date: Thu, 9 Feb 2006 10:24:21 -0600 20, 24 -- Message-ID: {BA876152E8653240BE8572E897083EE701849D33-at-UM-EMAIL09.um.umsystem.edu} 20, 24 -- X-MS-Has-Attach: 20, 24 -- X-MS-TNEF-Correlator: 20, 24 -- Thread-Topic: [Microscopy] Bacterial Pili 20, 24 -- thread-index: AcYtkwuBClshBOD6QsCkENhoqgFICQAAJ20Q 20, 24 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 20, 24 -- To: {hyi-at-emory.edu} 20, 24 -- Cc: {microscopy-at-microscopy.com} 20, 24 -- X-OriginalArrivalTime: 09 Feb 2006 16:24:22.0502 (UTC) FILETIME=[49041860:01C62D95] 20, 24 -- Content-Transfer-Encoding: 8bit 20, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19GOMnC014637 ==============================End of - Headers==============================
==============================Original Headers============================== 31, 31 -- From Edward.Calomeni-at-osumc.edu Thu Feb 9 11:18:04 2006 31, 31 -- Received: from pluto.osumc.edu (pluto.osumc.edu [140.254.120.27]) 31, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19HI3nX024586 31, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 11:18:03 -0600 31, 31 -- Received: from localhost (unknown [127.0.0.1]) 31, 31 -- by pfeg02.osumc.edu (Postfix) with ESMTP id 01EE2E22B 31, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 12:18:04 -0500 (EST) 31, 31 -- Received: from pfeg02.osumc.edu ([127.0.0.1]) 31, 31 -- by localhost (pfeg02 [127.0.0.1]) (amavisd-new, port 10024) with LMTP 31, 31 -- id 04255-01-25 for {Microscopy-at-microscopy.com} ; 31, 31 -- Thu, 9 Feb 2006 17:18:03 +0000 (GMT) 31, 31 -- Received: from msxc01.OSUMC.EDU (unknown [10.127.29.33]) 31, 31 -- by pfeg02.osumc.edu (Postfix) with ESMTP id D2A8BE03F 31, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 12:18:03 -0500 (EST) 31, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 31, 31 -- Content-class: urn:content-classes:message 31, 31 -- MIME-Version: 1.0 31, 31 -- Content-Type: text/plain; 31, 31 -- charset="iso-8859-1" 31, 31 -- Subject: RE: Bacterial Pili 31, 31 -- Date: Thu, 9 Feb 2006 12:18:03 -0500 31, 31 -- Message-ID: {7A541592F9A53A4E86E7A3D5C4C7F68C27F9F4-at-MSXC01} 31, 31 -- X-MS-Has-Attach: 31, 31 -- X-MS-TNEF-Correlator: 31, 31 -- Thread-Topic: RE: Bacterial Pili 31, 31 -- Thread-Index: AcYtleiHrKzoLX0fRmSx0dkDmP6AHwABibyw 31, 31 -- From: "Edward Calomeni" {Edward.Calomeni-at-osumc.edu} 31, 31 -- To: {Microscopy-at-microscopy.com} 31, 31 -- X-Virus-Scanned: amavisd-new at osumc.edu 31, 31 -- Content-Transfer-Encoding: 8bit 31, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19HI3nX024586 ==============================End of - Headers==============================
On Feb 9, 2006, at 4:45 AM, gerd.leitinger-at-meduni-graz.at wrote:
} We are just starting an experiment with en-bloc staining of tissue in } uranyl } acetate in 70% ethanol. } It seems to me that since the blocks are larger than thin sections, } they } probably take up more radioactive material- so should we take any } precautions when handling the blocks after the resin has cured? (i.e. } store } the blocks in separate special containers, wear gloves when sectioning } or } are there any special cleaning procedures for the knife?). } } thank you for sharing your knowledge } Dear Gerd, There is still a very small amount of radioactive material in the block; furthermore, uranium has a very long half-life, thus very small activity, and the alpha particles it emits have a short range, so any decays except those nearer the surface of the block than the thickness of the dead layer of your skin will not result in much radiation leaving the block. You can easily measure this with an ionization chamber (to detect the x- and gamma-radiation that make up most of the escaping radiation). That said, the laws regarding the handling of radioactive materials--even those with very little activity--vary from country to country, locale to locale, and sometimes for different institutions within a particular locale, so your safety office may mandate special procedures for storage and handling. 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 Feb 9 12:48:19 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19ImIdP002791 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 9 Feb 2006 12:48:19 -0600 4, 22 -- Received: from localhost (water-dog [192.168.1.26]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 33AB23422E 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 9 Feb 2006 10:48:18 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id 6844F33A1E 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 9 Feb 2006 10:48:17 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602091245.k19Cj91Q007324-at-ns.microscopy.com} 4, 22 -- References: {200602091245.k19Cj91Q007324-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {9fac5de4ab7271490d0c8faa23e1db2b-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] EM: precautions when en bloc staining 4, 22 -- Date: Thu, 9 Feb 2006 10:55:56 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Dear Hong, I have been using 2% Uranyl acetate to visualize bacterial pili.
I have been using 2% Uranyl acetate to visualize pili. I have had to play with the time of staining and rinsing in order to get a good contrast to visualize the pili on different bacterial genera. I have noticed that not all bacteria show the same distribution of pili. I have also noticed that the way you grow the bacteria also affect visualization of pili. For our bacteria I have noticed that I get far better results when I grow them up in stationary cultures without shaking them.
What I have been seeing it the bacteria behave differently when in aggregation versus solitude. This is more so owing to the several differnt kinds of pili each bacterial strain is capable of forming. If the bacteria you are working with produces different types of pili then you will see differences between the bacterial cells visualized on the same grid.
I have had the same strain of bacteria producing really small pili which were visualized only at a very high mag. Hope that was helpful regards, Vinod Graduate Student Dept Of Biology New Mexico State University On 2/9/06, hyi-at-emory.edu {hyi-at-emory.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 Microscopists: } } } We have a few users study bacterial pili using negative staining with } PTA (pH 6.5). We have got some pretty nice images. But often time, we } could not find pili even on the positive control. When we did see } pili, they were not on all bacteria in the same sample. Is this a } common phenomenon? Do bacteria lose their pili easily when the external } condition is not favorable? if that is the case, what should be done to } minimize the loss. } } Thank you in advance. } } Hong } Emory EM } } } ==============================Original Headers============================== } 5, 22 -- From hyi-at-emory.edu Thu Feb 9 10:06:53 2006 } 5, 22 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu } [170.140.8.222]) } 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19G6qLb005145 } 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 10:06:53 -0600 } 5, 22 -- Received: from [170.140.232.204] (localhost [127.0.0.1]) } 5, 22 -- by virginia.cc.emory.edu (8.13.4/8.13.4) with ESMTP id } k19G6qIx024009 } 5, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 11:06:52 -0500 } (EST) } 5, 22 -- Mime-Version: 1.0 (Apple Message framework v622) } 5, 22 -- Message-Id: {56be70af163d5629d409dc9e17804801-at-emory.edu} } 5, 22 -- Content-Type: text/plain; } 5, 22 -- charset=ISO-8859-1; } 5, 22 -- format=flowed } 5, 22 -- To: Microscopy-at-microscopy.com } 5, 22 -- From: Hong Yi {hyi-at-emory.edu} } 5, 22 -- Subject: Bacterial Pili } 5, 22 -- Date: Thu, 9 Feb 2006 11:06:51 -0500 } 5, 22 -- X-Mailer: Apple Mail (2.622) } 5, 22 -- X-imss-version: 2.037 } 5, 22 -- X-imss-result: Passed } 5, 22 -- X-imss-approveListMatch: *-at-emory.edu } 5, 22 -- Content-Transfer-Encoding: 8bit } 5, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k19G6qLb005145 } ==============================End of - Headers============================== }
==============================Original Headers============================== 5, 25 -- From nairvinods-at-gmail.com Thu Feb 9 13:08:22 2006 5, 25 -- Received: from wproxy.gmail.com (wproxy.gmail.com [64.233.184.202]) 5, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19J8Mrd012527 5, 25 -- for {microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 13:08:22 -0600 5, 25 -- Received: by wproxy.gmail.com with SMTP id i6so371281wra 5, 25 -- for {microscopy-at-microscopy.com} ; Thu, 09 Feb 2006 11:08:22 -0800 (PST) 5, 25 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 25 -- s=beta; d=gmail.com; 5, 25 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 5, 25 -- b=EaYibLaG48bwxRHn2D1rKg1YmKGyCmDnLAqNF+/f9UuSlcSOnBwQBCRPkBMXnmuuxJwTWlUo0r2HcibDDb78jLOk2qjdWIZ4b+Fwteoxowx/jA8BP9NFOLKwkpPzMMYrvgo4Ba23mGe3Ujr2MYVW72FvurUNj517iGySHrdgce8= 5, 25 -- Received: by 10.65.180.18 with SMTP id h18mr161301qbp; 5, 25 -- Thu, 09 Feb 2006 11:08:21 -0800 (PST) 5, 25 -- Received: by 10.64.203.13 with HTTP; Thu, 9 Feb 2006 11:08:21 -0800 (PST) 5, 25 -- Message-ID: {ea42a3900602091108occ4907aw88fdbc39652af5a0-at-mail.gmail.com} 5, 25 -- Date: Thu, 9 Feb 2006 12:08:21 -0700 5, 25 -- From: Vinod Nair {nairvinods-at-gmail.com} 5, 25 -- To: microscopy-at-microscopy.com 5, 25 -- Subject: Re: Bacterial Pili 5, 25 -- In-Reply-To: {200602091612.k19GC6Bt012799-at-ns.microscopy.com} 5, 25 -- MIME-Version: 1.0 5, 25 -- Content-Type: text/plain; charset=UTF-8 5, 25 -- Content-Disposition: inline 5, 25 -- References: {200602091612.k19GC6Bt012799-at-ns.microscopy.com} 5, 25 -- Content-Transfer-Encoding: 8bit 5, 25 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k19J8Mrd012527 ==============================End of - Headers==============================
I'm compiling a list of chemical stains/etches for delineating N+ and P+ source/drain implants/diffusions on semiconductor die cross sections. Any suggestions?
==============================Original Headers============================== 3, 22 -- From icmicroanalysis-at-cox.net Thu Feb 9 13:09:32 2006 3, 22 -- Received: from fed1rmmtao03.cox.net (fed1rmmtao03.cox.net [68.230.241.36]) 3, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19J9WI4014727 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 13:09:32 -0600 3, 22 -- Received: from officehp1 ([68.98.17.129]) by fed1rmmtao03.cox.net 3, 22 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with SMTP 3, 22 -- id {20060209190811.LEDO20875.fed1rmmtao03.cox.net-at-officehp1} 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 14:08:11 -0500 3, 22 -- From: "IC Microanalysis" {icmicroanalysis-at-cox.net} 3, 22 -- To: {Microscopy-at-microscopy.com} 3, 22 -- Subject: Stains/etches for delineating source/drains diffusions on semiconductor die cross sections 3, 22 -- Date: Thu, 9 Feb 2006 12:09:30 -0700 3, 22 -- Message-ID: {IKEHJCMBLNGNPJBGCPGJIEKACCAA.icmicroanalysis-at-cox.net} 3, 22 -- MIME-Version: 1.0 3, 22 -- Content-Type: text/plain; 3, 22 -- charset="iso-8859-1" 3, 22 -- Content-Transfer-Encoding: 7bit 3, 22 -- X-Priority: 3 (Normal) 3, 22 -- X-MSMail-Priority: Normal 3, 22 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2910.0) 3, 22 -- Importance: Normal 3, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
I put together a summary of the responses to my query on ESEM gas choices. People responded that they had experimented with Nitrogen, Nitrous Oxide, Oxygen, and Helium. Opinions were mixed on the performance of various gases (see below), but in general I feel comfortable trying Nitrogen in the near future. I received warnings that Helium can damage photomultipliers and Argon can damage ion pumps. Because of safety issues, we will most likely avoid oxygen and nitrous oxide for the time being.
FEI also contacted me and reminded that before experimenting with various gases in the ESEM, I should check with them to make sure that warranties are not voided by using any particular gas. If a gas could damage your system or components, the manufacturer should be able to tell you before you damage your system. Always check with the manufacturer before changing major system parameters.
My detector issue drew a number of separate questions and queries, and we are working separately with the manufacturers on this. We did learn that we can collect spectra with the SDD crystal at room temperature. This eliminates the potential for vapor condensation on the crystal surface. There is some peak broadening and a slightly higher background when we do this, but it does provide us with bulk data and may prove to be our final solution.
I also learned that many bulk gases also contain amounts of moisture that might condense under variable pressure conditions. Ultra high purity, low moisture gas, is sometimes considerably more expensive than bulk lab gas. I have to check with my vendor to find out what kind of moisture my tank might contain before hooking it to the ESEM.
Thanks again for everyone's help on this. The list has proven itself invaluable again.
Karl
----- "I have used water vapor, N2 and Argon. None of them gave as good a signal (image) as the water vapor did.... The Ionization of the other gasses is apparently not that good and give no nice images" ----- "I have been using dry nitrogen with my variable pressure instrument. We vent the system with the same gas. My EDS system seems to work well but have not run extensive tests to determine what effects the gas has other than the beam scattering issues." ----- " A disadvantage to using helium is your PMT's could be damaged, if it is released into the room. Also, another gas you could be careful with is argon, as it is a ion-pump poison (if released into a ion-pumped system). Nitrogen sounds friendly to me" ----- "In my experience, if you can't use oxygen the next best gas to use is argon - easily obtainable and provides good imaging conditions. Air and nitrogen should be avoided, since the nitrogen breaks down at too low a voltage and you can't get much signal. Some European groups looked at a wider range of imaging gases and nitrous oxide I believe did very well, although I have not tried it. I have tried oxygen and it works well, but tends to ruin the pump oil fairly quickly." ----- "The Quanta 200 uses a W filament. Unless it is a SFEG. If there are no ion pumps, you should be able to use Nitrogen or He. If there is an ion pump, do not use He. It will kill the pump.
I found that for VP (20-120Pa) that N2 works fine and is better than air (less vacuum issues). So, for ESEM, I would think that N2 ought to be the gas of choice as well." ----- " We routinely use He in our Hitachi VP-SEM. It has a tungsten gun and no ion pumps, so there is no problem with fouling those pumps as Gary Gaugler mentioned.
If you consider that He is a monoatomic species with a weight of 4 and that nitrogen is diatomic with a weight of 28, your gut can tell you that He will scatter less than N2 or room air. That is our experience and we can see the effect in iamges.
There is no effect on the x-ray signals from He that we have ever seen. He x-rays are below our low-energy discriminator. I suppose one could see N or O x-rays due to the gas, but I suppose their contribution is much less than from the sample. You could arrange a test to evaluate that, say you collected spectra from a beryllium or boron sample using high vacuum, helium, and nitrogen." ----- Original Post: --- ----
I am interested in hearing what type of gases people are using in their ESEM applications. We recently learned our silicon drift detector is incompatible with water vapor, and we apparently cannot complete any x-ray microanalysis while working in standard ESEM or VP modes (I was pretty surprised to learn this). We are thinking that using a dry gas such as nitrogen or helium will allow us to work in ESEM modes and use our x-ray system as well, as there is no potential for moisture condensing on the crystal surface.
Our ESEM is a Quanta 200, and it is not equipped with the peltier stage, so we mainly use the environmental modes to alleviate charging in uncoated samples. The x-ray system was purchased with the microscope about four years ago. I would rather not discuss the name of the x-ray vendor on the list, as we have a significant investment in this detector and do not foresee finding the money to purchase a new one soon.
Our first concern is the impact of the gas on spectra. We can coat samples and run them in high vacuum mode, but customers like spectra that represent the sample and not the coating material. We periodically have samples that cannot be coated, so ESEM becomes critical for our imaging needs. If the gas is going to have a dramatic impact on signal, are we better off coating and running high vacuum?
Is there a best choice for chamber gas? Our service engineer recommends nitrogen as having benefits for keeping the system clean. We can set it up fairly quickly, and I have a spare tank and regulator. I have heard of people using helium and believe that I read somewhere that there was some advantage to using helium.
I am still trying to get information from the vendor on whether this will allow us to use the x-ray and ESEM simultaneously, or whether we have options with different collimators or windows. Their responses have been pretty slow coming. _____
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 18, 23 -- From hagglundk1-at-nku.edu Fri Feb 10 10:10:59 2006 18, 23 -- Received: from mailfe2.nku.edu (exchange.nku.edu [192.122.237.68]) 18, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1AGAxYh024896 18, 23 -- for {microscopy-at-microscopy.com} ; Fri, 10 Feb 2006 10:10:59 -0600 18, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 18, 23 -- Fri, 10 Feb 2006 11:10:59 -0500 18, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 18, 23 -- Content-class: urn:content-classes:message 18, 23 -- MIME-Version: 1.0 18, 23 -- Content-Type: text/plain; 18, 23 -- charset="us-ascii" 18, 23 -- Subject: Follow up ESEM gas choices 18, 23 -- Date: Fri, 10 Feb 2006 11:10:56 -0500 18, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F358647-at-mailfac1.hh.nku.edu} 18, 23 -- X-MS-Has-Attach: 18, 23 -- X-MS-TNEF-Correlator: 18, 23 -- Thread-Topic: Follow up ESEM gas choices 18, 23 -- Thread-Index: AcYuXJL+d7H9dmpuRdS+OZyc5Rw51w== 18, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 18, 23 -- To: {microscopy-at-microscopy.com} 18, 23 -- X-OriginalArrivalTime: 10 Feb 2006 16:10:59.0208 (UTC) FILETIME=[94A0E880:01C62E5C] 18, 23 -- Content-Transfer-Encoding: 8bit 18, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1AGAxYh024896 ==============================End of - Headers==============================
Daniel, If you contact Gene Taylor at M. E. Taylor Engineering
Phone 301-774-6246
Fax 301-774-6711
www.semsupplies.com
I'm sure he can give you what you need. Scintillators have been his specialty for decades. Also, many of the other EM supply companies should be able to provide what you're looking for.
Disclaimer: A number of years ago when my business was larger, I was a distributor of Taylor supplies and have many happy customers.
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: Daniel.Salamon-at-nrc-cnrc.gc.ca [mailto:Daniel.Salamon-at-nrc-cnrc.gc.ca] Sent: Wednesday, February 08, 2006 4:35 PM To: kenconverse-at-qualityimages.biz
Hi everyone,
I will be experimenting with alternative scintillators for our Hitachi S4800 STEM unit. As I was advised to no touch the scintillator in the microscope, I hope someone will be able to offer measurements for the original S4800 scintillator.
Thanks, Daniel Salamon Technical Officer, Electron Microscopy
National Institute for Nanotechnology, NRC W6-017A ECERF Bldg, 9107-116 Street Edmonton, AB. T6G 2V4
==============================Original Headers============================== 5, 23 -- From Daniel.Salamon-at-nrc-cnrc.gc.ca Wed Feb 8 15:33:09 2006 5, 23 -- Received: from nrccenexf2.nrc.ca (nrccenexf2.nrc.ca [132.246.15.83]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k18LX8mG009977 5, 23 -- for {microscopy-at-microscopy.com} ; Wed, 8 Feb 2006 15:33:08 -0600 5, 23 -- Received: from nrccenexb2.nrc.ca ([132.246.15.98]) by nrccenexf2.nrc.ca with Microsoft SMTPSVC(6.0.3790.1830); 5, 23 -- Wed, 8 Feb 2006 16:33:07 -0500 5, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 23 -- Content-class: urn:content-classes:message 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; 5, 23 -- charset="us-ascii" 5, 23 -- Subject: S4800 STEM scintillator size 5, 23 -- Date: Wed, 8 Feb 2006 16:32:19 -0500 5, 23 -- Message-ID: {923687253229634E96E808B8D3124C83557517-at-nrccenexb2.nrc.ca} 5, 23 -- X-MS-Has-Attach: 5, 23 -- X-MS-TNEF-Correlator: 5, 23 -- Thread-Topic: S4800 STEM scintillator size 5, 23 -- Thread-Index: AcYs9yOtLSo8zGW/Rh2wOcOIwp9aqA== 5, 23 -- From: "Salamon, Daniel" {Daniel.Salamon-at-nrc-cnrc.gc.ca} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- X-OriginalArrivalTime: 08 Feb 2006 21:33:07.0447 (UTC) FILETIME=[4054A070:01C62CF7] 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k18LX8mG009977 ==============================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============================== 26, 24 -- From kenconverse-at-qualityimages.biz Fri Feb 10 10:20:24 2006 26, 24 -- Received: from qualityimages.biz (dpmail16.doteasy.com [65.61.209.16]) 26, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1AGKLGN000476 26, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Feb 2006 10:20:22 -0600 26, 24 -- Received: from Ken [68.64.242.101] by qualityimages.biz with ESMTP 26, 24 -- (SMTPD32-8.05) id ADB831560154; Fri, 10 Feb 2006 08:22:16 -0800 26, 24 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 26, 24 -- To: {Daniel.Salamon-at-nrc-cnrc.gc.ca} , 26, 24 -- "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 26, 24 -- Subject: RE: [Microscopy] S4800 STEM scintillator size 26, 24 -- Date: Fri, 10 Feb 2006 07:20:44 -0500 26, 24 -- Message-ID: {003b01c62e3c$746b43a0$6501a8c0-at-Ken} 26, 24 -- MIME-Version: 1.0 26, 24 -- Content-Type: text/plain; 26, 24 -- charset="us-ascii" 26, 24 -- X-Priority: 3 (Normal) 26, 24 -- X-MSMail-Priority: Normal 26, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 26, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 26, 24 -- Importance: Normal 26, 24 -- In-Reply-To: {200602082135.k18LZD0D015228-at-ns.microscopy.com} 26, 24 -- X-IMSTrailer: __IMail_7__ 26, 24 -- Content-Transfer-Encoding: 8bit 26, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1AGKLGN000476 ==============================End of - Headers==============================
I am posting the following ad for Dr. Jingyue Liu at Monsanto. Please contact Dr. Liu for further information, email address at the bottom of the request.
Lou Ross
Two Contract Researchers Are Needed in the Catalyst Characterization Group of Monsanto Company
Job Location: Creve Coeur Campus, St. Louis, Missouri 63167
The following two openings (one year contract researchers) are immediately available:
1) Research associate for TEM sample preparation. Required skills: extensive experience and expertise in ultramicrotoming thin sections of catalyst powders or other nanophase materials for transmission electron microscopy observation. This job requires strong hands-on skills and new method development for unique samples. Experience with sample preparation equipments such as high vacuum carbon coating, embedding, fixing and staining biological tissues, and maintenance of sample preparation facility is highly desirable. Experience in operating SEM instruments is a plus.
2) Research associate for catalyst preparation, treatment and characterization. Required skills: extensive experience in preparing model and practical catalysts or nanoparticles and TEM characterization of such samples. Experience in catalyst treatment and testing is a plus. Strong hands-on skills and demonstrated capability of designing complex experiments are required for this job.
Both jobs require extensive hands-on experiences in research labs. The following competencies are required: innovation in solving challenging problems; good communication and interpersonal skills; teamwork skills and results orientation.
Since Monsanto does not directly hire contract researchers, the selected candidates will work for a contract agency. Interested parties please send your resume and application letter to: Jingyue.liu-at-monsanto.com.
-- 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============================== 11, 17 -- From RossLM-at-missouri.edu Fri Feb 10 11:55:40 2006 11, 17 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 11, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1AHtd3Q012471 11, 17 -- for {microscopy-at-microscopy.com} ; Fri, 10 Feb 2006 11:55:39 -0600 11, 17 -- Received: from um-exproto9.um.umsystem.edu ([207.160.151.148]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 11, 17 -- Fri, 10 Feb 2006 11:55:39 -0600 11, 17 -- Received: from [128.206.78.231] ([128.206.78.231]) by um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft SMTPSVC(6.0.3790.1830); 11, 17 -- Fri, 10 Feb 2006 11:55:39 -0600 11, 17 -- Mime-Version: 1.0 11, 17 -- X-Sender: RossLM-at-pop.missouri.edu 11, 17 -- Message-Id: {p05200f30c0128247bb75-at-[128.206.78.231]} 11, 17 -- Date: Fri, 10 Feb 2006 11:55:37 -0600 11, 17 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 11, 17 -- From: Lou Ross {RossLM-at-missouri.edu} 11, 17 -- Subject: 2 contract positions at Monsanto in St. Louis 11, 17 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 11, 17 -- X-OriginalArrivalTime: 10 Feb 2006 17:55:39.0395 (UTC) FILETIME=[33E96530:01C62E6B] ==============================End of - Headers==============================
I am comtemplating broadening a mainly TEM undergrad course (with a lab) to include some scanning probe techniques. Does anyone know of reasonable texts which have some coverage?
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2220 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 email: L - marks -at- northwestern . edu http://www.numis.northwestern.edu -----------------------------------------------
==============================Original Headers============================== 4, 16 -- From ldmm-at-risc4.numis.northwestern.edu Fri Feb 10 12:25:19 2006 4, 16 -- Received: from risc4.numis.northwestern.edu (risc4.numis.northwestern.edu [129.105.122.70]) 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1AIPJKL022849 4, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Feb 2006 12:25:19 -0600 4, 16 -- Received: from localhost (ldmm-at-localhost) 4, 16 -- by risc4.numis.northwestern.edu (8.9.3 (PHNE_28760_binary)/8.9.3) with ESMTP id MAA24082 4, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Feb 2006 12:25:14 -0600 (CST) 4, 16 -- Date: Fri, 10 Feb 2006 12:25:14 -0600 (CST) 4, 16 -- From: LDM Microscopy {ldmm-at-risc4.numis.northwestern.edu} 4, 16 -- To: MSA listserver {Microscopy-at-microscopy.com} 4, 16 -- Subject: Texts that combine TEM + scanning probe techniques 4, 16 -- In-Reply-To: {000001c494e3$2098d330$b99bd280-at-paklabpgrover} 4, 16 -- Message-ID: {Pine.GHP.4.63.0602101223340.24076-at-risc4.numis.northwestern.edu} 4, 16 -- References: {000001c494e3$2098d330$b99bd280-at-paklabpgrover} 4, 16 -- MIME-Version: 1.0 4, 16 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (anna8261-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, February 10, 2006 at 02:52:59 ---------------------------------------------------------------------------
Email: anna8261-at-yahoo.com Name: anna naghshegar
Organization: pooysh
Education: Graduate College
Location: tehran, iran
Question: what is stain solution for double heterostructure InP/InGaAsP?
Hello listservers, I realize that this might be a silly question but I was wondering if there was anyone here who knows what the structure of colloidal gold is.
Thanks, Carlo
-- Carlo Franco Bolivar Balane
Box 4058, 222 Church Street, or Wolfe Laboratory Wesleyan University Station Rm. 157, HA Laboratories Middletown, CT, 06459-4058 Wesleyan University
==============================Original Headers============================== 7, 32 -- From cbalane-at-wesleyan.edu Sun Feb 12 01:38:59 2006 7, 32 -- Received: from post1.wesleyan.edu (post1.wesleyan.edu [129.133.6.131]) 7, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1C7cxSn031796 7, 32 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 01:38:59 -0600 7, 32 -- Received: from pony1.wesleyan.edu (pony1.wesleyan.edu [129.133.6.192]) 7, 32 -- by post1.wesleyan.edu (8.12.11/8.12.11) with ESMTP id k1C7dAPY015816 7, 32 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 02:39:10 -0500 7, 32 -- Received: from pony1.wesleyan.edu (pony1.wesleyan.edu [127.0.0.1]) 7, 32 -- by pony1.wesleyan.edu (8.12.11/8.12.11) with ESMTP id k1C7d045011036 7, 32 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 02:39:00 -0500 7, 32 -- Received: (from apache-at-localhost) 7, 32 -- by pony1.wesleyan.edu (8.12.11/8.12.11/Submit) id k1C7d0xP011034; 7, 32 -- Sun, 12 Feb 2006 02:39:00 -0500 7, 32 -- Received: from 129.133.127.145 7, 32 -- (SquirrelMail authenticated user cbalane); 7, 32 -- by webmail.wesleyan.edu with HTTP; 7, 32 -- Sun, 12 Feb 2006 02:39:00 -0500 (EST) 7, 32 -- Message-ID: {2201.129.133.127.145.1139729940.squirrel-at-129.133.127.145} 7, 32 -- Date: Sun, 12 Feb 2006 02:39:00 -0500 (EST) 7, 32 -- Subject: colloidal gold structure 7, 32 -- From: "Carlo Franco Balane-Bolivar" {cbalane-at-wesleyan.edu} 7, 32 -- To: Microscopy-at-microscopy.com 7, 32 -- User-Agent: SquirrelMail/1.4.3a-0.e3.1 7, 32 -- X-Mailer: SquirrelMail/1.4.3a-0.e3.1 7, 32 -- MIME-Version: 1.0 7, 32 -- Content-Type: text/plain;charset=iso-8859-1 7, 32 -- Content-Transfer-Encoding: 8bit 7, 32 -- X-Priority: 3 (Normal) 7, 32 -- Importance: Normal 7, 32 -- X-Wesleyan-MailScanner-Information: Please contact the ISP for more information 7, 32 -- X-Wesleyan-MailScanner: Found to be clean 7, 32 -- X-MailScanner-From: cbalane-at-wesleyan.edu ==============================End of - Headers==============================
In general multiply-twinned fcc domains I would have thought. Although depending on particle size, very small colloids can have structure forbidden by conventional bulk crystallography such as a 5-fold icosahedral or decahedral arrangements, plenty of literature around, both experimental and simulations. I seem to remember passivation of the gold 'cluster' can force the resulting colloid into certain structures. Generally Au over 3-4nm in diameter is mt-fcc though.
Neil Young Cluster Physics / Electron Microscopy Nanoscale Physics Research Laboratory School of Physics and Astronomy University of Birmingham UK
} Message Received: Feb 12 2006, 07:42 AM } From: cbalane-at-wesleyan.edu } To: neil-at-young8696.freeserve.co.uk } Cc: } Subject: [Microscopy] colloidal gold structure } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello listservers, } I realize that this might be a silly question but I was wondering if there } was anyone here who knows what the structure of colloidal gold is. } } Thanks, } Carlo } } } -- } Carlo Franco Bolivar Balane } } Box 4058, 222 Church Street, or Wolfe Laboratory } Wesleyan University Station Rm. 157, HA Laboratories } Middletown, CT, 06459-4058 Wesleyan University } } } } ==============================Original Headers============================== } 7, 32 -- From cbalane-at-wesleyan.edu Sun Feb 12 01:38:59 2006 } 7, 32 -- Received: from post1.wesleyan.edu (post1.wesleyan.edu [129.133.6.131]) } 7, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1C7cxSn031796 } 7, 32 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 01:38:59 -0600 } 7, 32 -- Received: from pony1.wesleyan.edu (pony1.wesleyan.edu [129.133.6.192]) } 7, 32 -- by post1.wesleyan.edu (8.12.11/8.12.11) with ESMTP id k1C7dAPY015816 } 7, 32 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 02:39:10 -0500 } 7, 32 -- Received: from pony1.wesleyan.edu (pony1.wesleyan.edu [127.0.0.1]) } 7, 32 -- by pony1.wesleyan.edu (8.12.11/8.12.11) with ESMTP id k1C7d045011036 } 7, 32 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 02:39:00 -0500 } 7, 32 -- Received: (from apache-at-localhost) } 7, 32 -- by pony1.wesleyan.edu (8.12.11/8.12.11/Submit) id k1C7d0xP011034; } 7, 32 -- Sun, 12 Feb 2006 02:39:00 -0500 } 7, 32 -- Received: from 129.133.127.145 } 7, 32 -- (SquirrelMail authenticated user cbalane); } 7, 32 -- by webmail.wesleyan.edu with HTTP; } 7, 32 -- Sun, 12 Feb 2006 02:39:00 -0500 (EST) } 7, 32 -- Message-ID: {2201.129.133.127.145.1139729940.squirrel-at-129.133.127.145} } 7, 32 -- Date: Sun, 12 Feb 2006 02:39:00 -0500 (EST) } 7, 32 -- Subject: colloidal gold structure } 7, 32 -- From: "Carlo Franco Balane-Bolivar" {cbalane-at-wesleyan.edu} } 7, 32 -- To: Microscopy-at-microscopy.com } 7, 32 -- User-Agent: SquirrelMail/1.4.3a-0.e3.1 } 7, 32 -- X-Mailer: SquirrelMail/1.4.3a-0.e3.1 } 7, 32 -- MIME-Version: 1.0 } 7, 32 -- Content-Type: text/plain;charset=iso-8859-1 } 7, 32 -- Content-Transfer-Encoding: 8bit } 7, 32 -- X-Priority: 3 (Normal) } 7, 32 -- Importance: Normal } 7, 32 -- X-Wesleyan-MailScanner-Information: Please contact the ISP for more information } 7, 32 -- X-Wesleyan-MailScanner: Found to be clean } 7, 32 -- X-MailScanner-From: cbalane-at-wesleyan.edu } ==============================End of - Headers============================== } }
==============================Original Headers============================== 6, 25 -- From neil-at-young8696.freeserve.co.uk Sun Feb 12 06:20:41 2006 6, 25 -- Received: from smtp2.freeserve.com (smtp2.wanadoo.co.uk [193.252.22.157]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1CCKd7F018464 6, 25 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 06:20:40 -0600 6, 25 -- Received: from me-wanadoo.net (localhost [127.0.0.1]) 6, 25 -- by mwinf3106.me.freeserve.com (SMTP Server) with ESMTP id 266D41C00085 6, 25 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 13:20:37 +0100 (CET) 6, 25 -- Received: from wwinf3103 (wwinf3103 [172.22.158.30]) 6, 25 -- by mwinf3106.me.freeserve.com (SMTP Server) with ESMTP id 205A51C00083 6, 25 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 13:20:37 +0100 (CET) 6, 25 -- X-ME-UUID: 20060212122037132.205A51C00083-at-mwinf3106.me.freeserve.com 6, 25 -- Message-ID: {30650493.1139746837118.JavaMail.www-at-wwinf3103} 6, 25 -- From: "Neil P. Young" {neil-at-young8696.freeserve.co.uk} 6, 25 -- Reply-To: neil-at-young8696.freeserve.co.uk 6, 25 -- To: Microscopy-at-microscopy.com 6, 25 -- Subject: RE: [Microscopy] colloidal gold structure 6, 25 -- Mime-Version: 1.0 6, 25 -- Content-Type: text/plain; charset=UTF-8 6, 25 -- Content-Transfer-Encoding: 7bit 6, 25 -- X-Originating-IP: [195.92.168.168] 6, 25 -- X-Wum-Nature: EMAIL-NATURE 6, 25 -- X-WUM-FROM: |~| 6, 25 -- X-WUM-TO: |~| 6, 25 -- X-WUM-REPLYTO: |~| 6, 25 -- Date: Sun, 12 Feb 2006 13:20:37 +0100 (CET) ==============================End of - Headers==============================
Karl, I hadn't seen any answer to your question so I'll give it a shot. For those of us who grew up with Be windows, K bigger than L seems normal. However, the norm for light element detectors is the opposite, at least if you normally operate at 20kV or so. I'm wondering if you've had a long-standing contamination problem with your light element window. Now that it's clean, you may be seeing the correct ratio and may find that your light element detection is also better.
The other thing to be certain of is that you are operating at the same kV as you were earlier. Lower kV will favor lower energy peaks and higher kV will favor higher energy peaks. It can be quite dramatic. Put a little graphite on a piece of Cu tape and collect a spectrum from an area containing both. Maintain a constant dead-time while collecting a spectrum at 30 kV and another at about 2 kV. You'd never know it's the same sample.
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: hagglundk1-at-nku.edu [mailto:hagglundk1-at-nku.edu] Sent: Tuesday, January 24, 2006 10:33 AM To: kenconverse-at-qualityimages.biz
I have a colleague who recently experienced some down time on his SEM. The system went through repeated cycles of pump down, and venting and was eventually left powered down while waiting on a computer replacement.
After getting everything operating again, an oil film was found that had built up on the x-ray detector thin window. This was initially causing an unacceptable amount of background in the 0-3kV range of the x-ray signal as well as an overall low count rate. The collimator was removed and cleaned, which corrected the noise and count rate. They are in the process of replacing the roughing lines and cleaning the chamber to prevent further contamination. Now, when calibrating with a copper standard, the ratio of the L line signal versus the K lines is dramatically favoring the L. This was not the case before.
Any ideas why this might be happening and how to correct it?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 5, 23 -- From hagglundk1-at-nku.edu Tue Jan 24 09:02:19 2006 5, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k0OF2JlW001861 5, 23 -- for {microscopy-at-microscopy.com} ; Tue, 24 Jan 2006 09:02:19 -0600 5, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 5, 23 -- Tue, 24 Jan 2006 10:02:19 -0500 5, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 5, 23 -- Content-class: urn:content-classes:message 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; 5, 23 -- charset="us-ascii" 5, 23 -- Subject: SEM x-ray peak ratios 5, 23 -- Date: Tue, 24 Jan 2006 10:02:18 -0500 5, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3585FA-at-mailfac1.hh.nku.edu} 5, 23 -- X-MS-Has-Attach: 5, 23 -- X-MS-TNEF-Correlator: 5, 23 -- Thread-Topic: SEM x-ray peak ratios 5, 23 -- Thread-Index: AcYg9yuMm942rGb1QfKCevfjxyMD/g== 5, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- X-OriginalArrivalTime: 24 Jan 2006 15:02:19.0222 (UTC) FILETIME=[2BE72F60:01C620F7] 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k0OF2JlW001861 ==============================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============================== 22, 23 -- From kenconverse-at-qualityimages.biz Sun Feb 12 14:39:26 2006 22, 23 -- Received: from qualityimages.biz (dpmail16.doteasy.com [65.61.209.16]) 22, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1CKdPhn012116 22, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 12 Feb 2006 14:39:26 -0600 22, 23 -- Received: from Ken [68.64.242.101] by qualityimages.biz with ESMTP 22, 23 -- (SMTPD32-8.05) id AD6DA2CE00B8; Sun, 12 Feb 2006 12:41:17 -0800 22, 23 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 22, 23 -- To: {hagglundk1-at-nku.edu} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 22, 23 -- Subject: RE: [Microscopy] SEM x-ray peak ratios 22, 23 -- Date: Sun, 12 Feb 2006 15:38:54 -0500 22, 23 -- Message-ID: {001701c63014$5fed1550$6501a8c0-at-Ken} 22, 23 -- MIME-Version: 1.0 22, 23 -- Content-Type: text/plain; 22, 23 -- charset="us-ascii" 22, 23 -- X-Priority: 3 (Normal) 22, 23 -- X-MSMail-Priority: Normal 22, 23 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 22, 23 -- Importance: Normal 22, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 22, 23 -- In-Reply-To: {200601241533.k0OFX9f9007423-at-ns.microscopy.com} 22, 23 -- X-IMSTrailer: __IMail_7__ 22, 23 -- Content-Transfer-Encoding: 8bit 22, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1CKdPhn012116 ==============================End of - Headers==============================
A colleague has a Nikon SMZ-2T Microscope that he is trying to rescue. He is after a copy of the manual. If anyone can help pls contact him at hbeh-at-hotmail.com or via myself.
Thank you for your help
Regards George
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==============================Original Headers============================== 8, 27 -- From George.Theodossiou-at-amcor.com.au Sun Feb 12 18:58:09 2006 8, 27 -- Received: from aiti251.amcor.com.au (mail.amcor.com.au [202.14.180.248] (may be forged)) 8, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1D0w78V024534 8, 27 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Feb 2006 18:58:08 -0600 8, 27 -- Received: from aadcex0002.amcor.net (unverified) by aiti251.amcor.com.au 8, 27 -- (Content Technologies SMTPRS 4.3.19) with ESMTP id 8, 27 -- {T766fb3d9f0a0de98c8538-at-aiti251.amcor.com.au} for 8, 27 -- {Microscopy-at-microscopy.com} ; Mon, 13 Feb 2006 12:03:18 +1100 8, 27 -- Received: from aadcex0004.amcor.net ([160.222.80.235]) by 8, 27 -- aadcex0002.amcor.net with Microsoft SMTPSVC (6.0.3790.211); Mon, 13 Feb 8, 27 -- 2006 11:58:05 +1100 8, 27 -- Received: from 160.222.214.35 ([160.222.214.35]) by aadcex0004.amcor.net 8, 27 -- ([160.222.80.235]) with Microsoft Exchange Server HTTP-DAV; Mon, 13 Feb 8, 27 -- 2006 00:58:05 +0000 8, 27 -- User-Agent: Microsoft-Entourage/11.2.1.051004 8, 27 -- Date: Mon, 13 Feb 2006 11:57:31 +1100 8, 27 -- Subject: Nikon Microscope 8, 27 -- From: "George.Theodossiou" {George.Theodossiou-at-amcor.com.au} 8, 27 -- To: {Microscopy-at-microscopy.com} 8, 27 -- Message-ID: {C01624AB.8E6%George.Theodossiou-at-Amcor.com.au} 8, 27 -- Thread-Topic: Nikon Microscope 8, 27 -- Thread-Index: AcYwOHehtgRappwrEdqDLgANkzYUMg== 8, 27 -- Mime-version: 1.0 8, 27 -- Content-type: text/plain; charset="US-ASCII" 8, 27 -- Content-transfer-encoding: 7bit 8, 27 -- X-OriginalArrivalTime: 13 Feb 2006 00:58:06.0019 (UTC) 8, 27 -- FILETIME=[8C80C930:01C63038] ==============================End of - Headers==============================
Thank you everybody who replied to my question regarding safety measures when working with uranyl acetate stained blocks.
To sum up the answers: The binding capacity of tissue to uranyl acetate is apparently low and therefore very little (if any) radioactivity can escape the blocks. However, when trimming I have been advised to wear gloves and a mask to prevent myself from inhaling chips from the specimen, and it seems necessary that we collect the chips and dispose of them as radioactive waste.
thank you
Gerd
Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
We are trying to viusalize infected erythrocytes (malarial parasite) on confocal microscope by indirect fluorescence without any fixation and washing is carried out with PBS
The immages are not satisfactory because fading/bleaching is fast though we are using % DABCO in 50% glycerol and secondly lot of background noise.
Most of the literature shows the fixation with ethanol/methenol but I would like to carry out the fixation with paraformaldehyde for the obvious reason and washing with MSM-PIPES with tris.
any suggestion???? or any body can describle the sample preparation for infected erythrocytes using paraformaldehy as a fixative and MSM PIPES as a washing buffer...
Does non fixation of specimen play any role in fast bleaching???
Regards Shrunali Scientist IMTECH, India
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==============================Original Headers============================== 10, 18 -- From aarti_harle-at-yahoo.co.in Mon Feb 13 04:53:08 2006 10, 18 -- Received: from web8308.mail.in.yahoo.com (web8308.mail.in.yahoo.com [202.43.219.220]) 10, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1DAr74m018713 10, 18 -- for {Microscopy-at-Microscopy.com} ; Mon, 13 Feb 2006 04:53:07 -0600 10, 18 -- Received: (qmail 90502 invoked by uid 60001); 13 Feb 2006 10:53:05 -0000 10, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 18 -- s=s1024; d=yahoo.co.in; 10, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 10, 18 -- b=XV1GTCKSYckbhHBodcI+9aRMHIyL3k7aSk/fx0siVLqY2GubzN4t3ETtsQ5SvUNHdvo0HZCbwY0LNwqIEVo5wG0A2w5hFGhRK2sdU4Sw5T+v7Wv8JD5Q0rRO14w39oy6TAwfWGnMr0IXewk2yCdWRCswRSZSwMNHWt722M1sryM= ; 10, 18 -- Message-ID: {20060213105305.90500.qmail-at-web8308.mail.in.yahoo.com} 10, 18 -- Received: from [203.197.210.215] by web8308.mail.in.yahoo.com via HTTP; Mon, 13 Feb 2006 02:53:05 PST 10, 18 -- Date: Mon, 13 Feb 2006 02:53:05 -0800 (PST) 10, 18 -- From: Aarti Harle {aarti_harle-at-yahoo.co.in} 10, 18 -- Subject: confocal microscopy 10, 18 -- To: Microscopy-at-Microscopy.com 10, 18 -- MIME-Version: 1.0 10, 18 -- Content-Type: text/plain; charset=iso-8859-1 10, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
You don't specify which fluorochrome you are using. If you are using FITC or Rhodamine, that is one of the reasons you are getting rapid fading. The Alexa488 and Alexa568 fluorochromes from Molecular Probes (Invitrogen) are far superior in this regard - especially for confocal. Molecular Probes also has a new anti-fade mounting medium called Prolong Gold but I don't have enough experience at the moment to discuss its efficacy. Fixation will not have any effect on bleaching but will contribute to background noise (especially glutaraldehyde). Generally 2% PF is okay in regards to background fluorescence and sometimes it is safe to add 0.1 - 02% glutaraldehyde. Aldehyde fixatives may, however, interfere with your antibody recognizing its epitope. good luck.
rote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I'm working on getting a Zeiss 940A up and running for my local high school. We are working on a shoe-string budget as one may expect from a small public high school.
I am looking for technical documentation for this instrument so that I can try and get it functioning. If anyone has schematics or any other technical documentation for instrument this I would greatly appreciate getting copies in some way.
If anyone has a similar instrument that would be able to donate it to us for a spare parts machine, we would come to pick it up. We are located in the Hudson Valley.
I am willing to pay for copies if need be, as well as transportation costs.
Thank you in advance,
dj
==============================Original Headers============================== 7, 22 -- From dljones-at-bestweb.net Mon Feb 13 09:22:44 2006 7, 22 -- Received: from smtp2.bestweb.net (smtp2.bestweb.net [209.94.103.44]) 7, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1DFMigH008985 7, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 13 Feb 2006 09:22:44 -0600 7, 22 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) 7, 22 -- by smtp2.bestweb.net (Postfix) with ESMTP id 0659D1CD06 7, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 13 Feb 2006 10:22:42 -0500 (EST) 7, 22 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) 7, 22 -- by smtp2.bestweb.net (Postfix) with ESMTP id 9164D1CCCB 7, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 13 Feb 2006 10:22:41 -0500 (EST) 7, 22 -- Date: Mon, 13 Feb 2006 10:31:56 -0500 (Eastern Standard Time) 7, 22 -- From: "David L. Jones" {dljones-at-bestweb.net} 7, 22 -- To: Microscopy-at-microscopy.com 7, 22 -- Subject: Zeiss 940A 7, 22 -- Message-ID: {Pine.WNT.4.62.0602131022110.1512-at-dlj} 7, 22 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 7, 22 -- MIME-Version: 1.0 7, 22 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 7, 22 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net 7, 22 -- X-Spam-Level: 7, 22 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed 7, 22 -- version=3.0.2 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (roncastellano-at-adelphia.net) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, February 13, 2006 at 13:21:29 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both roncastellano-at-adelphia.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: roncastellano-at-adelphia.net Name: Ron Castellano
Organization: Wolfram Analytical Labs
Education: Undergraduate College
Location: 924 Fords Corner Road, Nanty Glo, PA 15943
Title: Assistance to setup a JOEL JSM-35C SEM
Question: I'm a retired assay technician(precious metal analysis) I have purchased an old SEM for private use in a small lab I am building here. I need someone to set up and test unit at my site. The unit is said to be operational when de-installed, it appears to be in good shape. Are there any technicians, perhaps a graduate student, who can assist me. Of course I am willing to pay any reasonable fee for this service. Ron Castellano
I am in need of a working video card for a Gatan 673 Wide TV Camera, vintage 1987. This camera system is used on a Philips CM12 for teaching purposes. The video card model is Schlumberger/Fairchild FAB-2-1-28 rev. F.
I will consider buying the controller if you do not wish to sell the card separately.
If you wish you can contact me offline.
Thanks in advance,
Fred Pearson
******************************************* Fred Pearson McMaster University Brockhouse Institute for Materials Research ABB-B145 1280 Main Street West Hamilton ON. Canada L8S 4M1
Lehigh University, Center for Advanced Materials and Nanotechnology, Bethlehem, PA is offering for sale a JEOL-6300F FEGSEM which will be decommissioned in mid-March 2006. The Microscope includes an Oxford/ISIS thin window EDS system, JEOL dry airlock and ARC64 digital imaging system, solid state backscatter detector, IR chamberview camera. Asking price: $60K Interested parties can contact Dr. Chris Kiely at chk5-at-lehigh.edu.
For technical questions regarding the microscope please contact Bill Mushock wim5-at-lehigh.edu
--
==============================Original Headers============================== 6, 22 -- From wim5-at-lehigh.edu Wed Feb 15 12:43:09 2006 6, 22 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 6, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1FIh93u029868 6, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Feb 2006 12:43:09 -0600 6, 22 -- Received: from lehigh.edu (Dyn055133.mat.Lehigh.EDU [128.180.55.133]) 6, 22 -- (authenticated bits=0) 6, 22 -- by rain.CC.Lehigh.EDU (8.13.5/8.13.5) with ESMTP id k1FIh9Uv018394 6, 22 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 6, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Feb 2006 13:43:09 -0500 6, 22 -- Message-ID: {43F37517.5020702-at-lehigh.edu} 6, 22 -- Date: Wed, 15 Feb 2006 13:38:15 -0500 6, 22 -- From: William J Mushock {wim5-at-lehigh.edu} 6, 22 -- Organization: Lehigh University 6, 22 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 6, 22 -- X-Accept-Language: en,en-US 6, 22 -- MIME-Version: 1.0 6, 22 -- To: Microscopy-at-microscopy.com 6, 22 -- Subject: JEOL JSM-6300f FEGSEM for sale 6, 22 -- Content-Type: text/plain; charset=us-ascii; format=flowed 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- X-Virus-Scanned: ClamAV version 0.88, clamav-milter version 0.87 on rain.CC.Lehigh.EDU 6, 22 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I'm writing this as a response to Chris Kiely's post, but it is a very general point that probably bears repeating from time to time.
If a piece of equipment has been purchased by US Government funds (regardless of agency), then it is not permissible to use other US Government funds to re-purchase the same piece of equipment later. This is true even if title to the equipment has been passed to the holding instrumentation.
To use the specific example of the Lehigh SEM, if it was purchased through a government grant, I would not be able to buy it from them, regardless of how much use it would be to me, because the only funds I have available (at least for the purposes of my illustration!) are NSF funds. This is a limitation enforced on me, as the spender of Government funds, rather than on the seller (for they are - presumably - selling their own property), but it does mean that I need to know up front whether the instrument was purchased with any US Government funds (even if other funds were used as well) before I can decide whether I might be interested in it. I, and my institution, would be in major hot water should an auditor discover I had used government funds in this way (I suspect that here at MIT our internal systems would catch this before the sale went through, but that may not be true everywhere).
Tony Garratt-Reed.
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*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Because it's abstract time for both SCANNING 2006 and M&M, we know there is time pressure on all contributors. We are therefore extending the abstract deadline for SCANNING 2006 to March 1. Abstracts will appear in the Program Issue of SCANNING (March-April Issue) if received by March 1. Thank you.
For current program information and to download the meeting and hotel registration forms, please visit www.scanning.org.
This Question was submitted to Ask-A-Microscopist by (grmitch-at-netzero.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, February 15, 2006 at 20:08:01 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both grmitch-at-netzero.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: grmitch-at-netzero.com Name: Linda Mitchell
Organization: BPS Environmental Center
Education: K-8 Grade Grammar School
Location: Birmingham, Michigan USA
Title: Microorganisms!
Question: I will be teaching a unit on microscopic life to 5th graders next month, March (quite a cold month here in Michigan). My question for you: What is the best way to keep my microorganisms alive throughout the program? We obtain our samples directly from the 10 acres of the property here at the nature center. One of the samples is from our pond area and the second from the swamp woods area. We have no difficulty in finding a healthy sample, yet the problem that we often encounter is keeping them alive. We have supplies - lab pans, microscopes, even an aquarium that is our "indoor pond" when the water ices over. Do you have any feeding, climate tips that would help in these areas? This is a program that is enjoyed by both the staff and students - they are so amazed by what they find. Thanks for your input.
I'm out of the country right now, but I have been getting multiple questions about problems with the MM2006 server for submitting papers.
Please note that there was a HUGE spike in the paper submissions and the server is overloaded. As a consequence the MM2006 Executive committee has extended the deadline for paper submissions.
A note to this effect is appended below.
Nestor Your Friendly Neighborhood SysOp (in Australia for the moment).
The paper submission deadline for M&M 2006 in Chicago has been extended two days until 5:00pm Pacific Standard Time on Friday February 17, 2006. Due to an ongoing problem with the server today we have decided to extend the deadline to allow everyone the opportunity to submit their papers. Please try to access the paper submission site {http://bono.cup.org/} http://bono.cup.org/ later to register and submit your paper. Our sincerest apologies for any problems this may have caused.
Thanks, Paul Kotula Microscopy &Microanalysis 2006 Program Chair
Paul G. Kotula, Ph.D. Principal Member of Technical Staff Materials Characterization Department Sandia National Laboratories PO Box 5800, MS 0886 Albuquerque, NM 87185-0886
ph:(505) 844-8947
==============================Original Headers============================== 12, 11 -- From zaluzec-at-microscopy.com Thu Feb 16 14:52:48 2006 12, 11 -- Received: from [203.33.121.155] (msdvpn28.msd.anl.gov [130.202.238.92]) 12, 11 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1GKqh4n008778 12, 11 -- for {microscopy-at-microscopy.com} ; Thu, 16 Feb 2006 14:52:45 -0600 12, 11 -- Mime-Version: 1.0 12, 11 -- Message-Id: {p06020404c01a95b6217e-at-[203.33.121.155]} 12, 11 -- Date: Fri, 17 Feb 2006 07:52:41 +1100 12, 11 -- To: microscopy-at-microscopy.com 12, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 12, 11 -- Subject: MM2006 Server OverLoad - Deadline extended 12, 11 -- 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 sue.tyler-at-noaa.gov as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: sue.tyler-at-noaa.gov Name: Sue Tyler
Organization: NOAA
Title-Subject: [Filtered] Glass knife breaker
Question: I read the recent string on the glass knife makers and I was interested in the conclusion. We are interested in purchasing either the GKM or the Leica KMR. Any pros or cons would be appreciated.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both diller-at-stefan-diller.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Organization: Scientific Photography
Title-Subject: [Filtered] Tracor Northern TN2000 EDS system
Question: Dear All, is anybody out there knowing some details or having some images available of an 1991 Tracor Northern TN2000 EDS system?
Is there still a possiblity to get service in Germany? Is there any possibility to get the EDS data out of the analyzer into a PC for saving, printing, emailing? What is the latest version of software for the TN2000?
Please reply offline, if convenient... I will do a summarizing for the list...
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 soleimanij-at-tbzmed.ac.ir as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: soleimanij-at-tbzmed.ac.ir Name: Jafar Soleimani Rad
Organization: University
Title-Subject: [Filtered] cutting a polymer
Question: We are having problem with cutting a polymer, Acrylonitril Butadien Styrene (ABS. After embedding in resin it dose not stick to it and becomes separated when trying to cut with ultramicrotom. It is also impossible to cut paraffin blocks. I would appreciate if you guide me with your experiences in this matter.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both b.farwell-at-unf.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: We are using a Molecular Imaging AFM and controller and are using "MI Metrology Series 2000 AFM System" program to interface to the AFM controller. The computer is having connection issues with the controller. Cables have been checked, and the setup disk has run on the controller, it gives 4 beeps, however the computer still can't connect. Ping gives no response. Anybody have any ideas? Thanks very much Brenton
I recently acquired a used Balzers MED 010 of unknown vintage. I am in need of the User Manual, or any such literature. I would like to get this unit back to operating condition.
Thanks,
John Crum NCMIR UCSD
==============================Original Headers============================== 4, 18 -- From jcrum-at-ncmir.ucsd.edu Thu Feb 16 15:52:19 2006 4, 18 -- Received: from ncmir.ucsd.edu (ncmir.ucsd.edu [132.239.16.23]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1GLqIWF027723 4, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Feb 2006 15:52:19 -0600 4, 18 -- Received: from [192.168.10.237] (capisce [132.239.16.109]) 4, 18 -- by ncmir.ucsd.edu (8.11.7p1+Sun/8.11.7) with ESMTP id k1GLqIL04880 4, 18 -- for {Microscopy-at-Microscopy.Com} ; Thu, 16 Feb 2006 13:52:18 -0800 (PST) 4, 18 -- Message-ID: {43F4F412.1060004-at-ncmir.ucsd.edu} 4, 18 -- Date: Thu, 16 Feb 2006 13:52:18 -0800 4, 18 -- From: John Crum {jcrum-at-ncmir.ucsd.edu} 4, 18 -- Organization: University of California San Diego CRBS/NCMIR 4, 18 -- User-Agent: Mozilla Thunderbird 1.0.2 (Macintosh/20050317) 4, 18 -- X-Accept-Language: en-us, en 4, 18 -- MIME-Version: 1.0 4, 18 -- To: Microscopy-at-microscopy.com 4, 18 -- Subject: Wanted: Balzers MED 010 Manual 4, 18 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am looking for a used PSEM (or parts) to purchase. Preferably this system would be the original model manufactured by RJ Lee Instruments in the mid to late 90’s. Anyone that can help me out, please email directly to andy-at-psemdoctor.com. Thank you,
Andrew L. Zobel
PSEMDOCTOR, LLC 117 Bryant Dr. Pittsburgh, PA 15235 Tel. 412-215-8906 Fax 412-241-3598 WWW.PSEMDOCTOR.COM
==============================Original Headers============================== 6, 22 -- From andy-at-psemdoctor.com Thu Feb 16 17:20:44 2006 6, 22 -- Received: from rwcrmhc14.comcast.net (rwcrmhc14.comcast.net [204.127.192.84]) 6, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1GNKinN025744 6, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Feb 2006 17:20:44 -0600 6, 22 -- Received: from azdesktop (c-24-3-48-75.hsd1.pa.comcast.net[24.3.48.75]) 6, 22 -- by comcast.net (rwcrmhc14) with SMTP 6, 22 -- id {20060216232043m1400qhsbqe} ; Thu, 16 Feb 2006 23:20:43 +0000 6, 22 -- Reply-To: {andy-at-psemdoctor.com} 6, 22 -- From: "Andrew L. Zobel" {andy-at-psemdoctor.com} 6, 22 -- To: {Microscopy-at-microscopy.com} 6, 22 -- Subject: [Microscopy] Looking for used PSEM 6, 22 -- Date: Thu, 16 Feb 2006 18:20:43 -0500 6, 22 -- Organization: PSEMDOCTOR, LLC 6, 22 -- Message-ID: {000701c6334f$9c923ba0$6501a8c0-at-azdesktop} 6, 22 -- MIME-Version: 1.0 6, 22 -- Content-Type: text/plain; 6, 22 -- charset="iso-8859-1" 6, 22 -- X-Mailer: Microsoft Office Outlook 11 6, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 6, 22 -- Thread-Index: AcYzTv46OmN0PyDUSIGoUkBwqvLP/AAAFumw 6, 22 -- Content-Transfer-Encoding: 8bit 6, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1GNKinN025744 ==============================End of - Headers==============================
Hi Sue We have the Leica glass knife breaker since 2001. We are very happy with it. Reliable, good knives! Our users like it in preference to our old workhorse LKB knife breakers. Elaine
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-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada (2003-2005) University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
==============================Original Headers============================== 4, 29 -- From ech-at-interchange.ubc.ca Thu Feb 16 19:56:45 2006 4, 29 -- Received: from mta1.mail-relay.ubc.ca (mta1.mail-relay.ubc.ca [137.82.45.2]) 4, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1H1uiLA004198 4, 29 -- for {microscopy-at-microscopy.com} ; Thu, 16 Feb 2006 19:56:44 -0600 4, 29 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 4, 29 -- by mta1.mail-relay.ubc.ca (8.12.11/8.12.11) with ESMTP id k1H1ufaE023680 4, 29 -- for {microscopy-at-microscopy.com} ; Thu, 16 Feb 2006 17:56:41 -0800 (PST) 4, 29 -- (envelope-from ech-at-interchange.ubc.ca) 4, 29 -- Received: from [137.82.85.193] (echpowerbook.emlab.ubc.ca [137.82.85.193]) 4, 29 -- by smtp.interchange.ubc.ca 4, 29 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 4, 29 -- with ESMTPA id {0IUT00CU86QGQU-at-smtp.interchange.ubc.ca} for 4, 29 -- microscopy-at-microscopy.com; Thu, 16 Feb 2006 17:56:41 -0800 (PST) 4, 29 -- Date: Thu, 16 Feb 2006 17:56:38 -0800 4, 29 -- From: Elaine Humphrey {ech-at-interchange.ubc.ca} 4, 29 -- Subject: Re: [Microscopy] viaWWW: Glass Knife Breaker 4, 29 -- In-reply-to: {200602162058.k1GKw9iM025479-at-ns.microscopy.com} 4, 29 -- X-Sender: ech-at-mail.interchange.ubc.ca 4, 29 -- To: microscopy-at-microscopy.com 4, 29 -- Cc: sue.tyler-at-noaa.gov 4, 29 -- Message-id: {a06100504c01adcc79da1-at-[137.82.85.193]} 4, 29 -- MIME-version: 1.0 4, 29 -- Content-type: text/plain; format=flowed; charset=us-ascii 4, 29 -- References: {200602162058.k1GKw9iM025479-at-ns.microscopy.com} 4, 29 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.02.16.164605 4, 29 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 4, 29 -- X-PerlMx-Spam: Probability=7%, Report=IP_HTTP_ADDR 0, __C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0 4, 29 -- X-Spam-Level: 4, 29 -- X-Spam-Flag: No ==============================End of - Headers==============================
A colleague is looking for the instruction manual for a Bausch & Lomb Research I metallograph. If anybody can help him in this quest please respond directly to him, Gregory Dexter at greg-at-met-sol.com .
Thanks,
Jeff Stewart Metallographic Laboratory Manager Stern-Leach Company 49 Pearl Street Attleboro, MA 02703 508-222-7400 x1329
==============================Original Headers============================== 5, 21 -- From jeff-at-metallography.com Fri Feb 17 08:43:23 2006 5, 21 -- Received: from Q1-ABI-TX.sanimail.com (mail6.mailsystem.us [67.97.234.238]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HEhM02018323 5, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 08:43:23 -0600 5, 21 -- Received: (qmail 68083 invoked by uid 1010); 17 Feb 2006 15:00:57 -0000 5, 21 -- Received: from unknown (HELO jstewart) (4.154.213.130) 5, 21 -- by mail6.mailsystem.us with SMTP; 17 Feb 2006 15:00:57 -0000 5, 21 -- Message-ID: {001801c633d0$9895b500$959610ac-at-sternleach.com} 5, 21 -- Reply-To: "Jeff Stewart" {jeff-at-metallography.com} 5, 21 -- From: "Jeff Stewart" {jeff-at-metallography.com} 5, 21 -- To: {Microscopy-at-microscopy.com} 5, 21 -- Subject: Seeking manual for B&L Research I metallograph 5, 21 -- Date: Fri, 17 Feb 2006 09:41:06 -0500 5, 21 -- MIME-Version: 1.0 5, 21 -- Content-Type: text/plain; 5, 21 -- charset="iso-8859-1" 5, 21 -- Content-Transfer-Encoding: 7bit 5, 21 -- X-Priority: 3 5, 21 -- X-MSMail-Priority: Normal 5, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1409 5, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 ==============================End of - Headers==============================
For years we have been wondering where the dark material was coming from that accumulated in our water filters. These are filters in the closed circuit lines between our microscopes and water recirculating units. The lines are mainly in the ceiling or totally insulated as they run down the walls in the scope rooms. Imaging our surprise when we went to do a minor repair on one and watched as the plumber removed a regulator and inserted a piece of galvanized pipe.
Apparently when the building was built (20 years ago), the contractor used galvanized pipe when copper had been specified. As it was hidden, we did not know about the switch. All visible lines hooking up the chiller compressor cooling with building water were copper.
Well now these galvanized lines are really breaking down and clogging the water pumps. We intend to replace all but were questioning whether it would be best to replace with copper or PVC piping. Any suggestions?
One concern was whether there would be the need to acid clean these lines in the future (this is done routinely to the compressor lines to remove mineral build-up). Since it is closed circuit, we should not accumulate large amounts of minerals even though tap or deionized water will be used for the system. We also can control algae growth with chemicals. Any suggestions on this and should this dictate which material is used for the pipes?
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 Feb 17 11:51:22 2006 8, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 8, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HHpMgE030390 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 11:51:22 -0600 8, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 8, 21 -- Fri, 17 Feb 2006 12:51:21 -0500 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, 17 Feb 2006 17:51:21 +0000 8, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 8, 21 -- Date: Fri, 17 Feb 2006 12:51:20 -0500 8, 21 -- Subject: Microscope cooling lines 8, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 8, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 8, 21 -- Message-ID: {C01B7748.D5AB%dsherman-at-purdue.edu} 8, 21 -- Thread-Topic: Microscope cooling lines 8, 21 -- Thread-Index: AcYz6sIxAPacp5/eEdqwBAARJN08Mg== 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: 17 Feb 2006 17:51:21.0666 (UTC) FILETIME=[C32F5220:01C633EA] ==============================End of - Headers==============================
I wonder if anyone has a manual for the Denton DV-502A vacuum evaporator that I can borrow. Ours was lost during moving to storage and now I have to set it up again. It would be nice to make sure I set it up correctly. Denton can supply the manual for $250 so I am looking at other less expensive ways first. Thanks a lot!
-- Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519) The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
==============================Original Headers============================== 3, 19 -- From wpchan-at-u.washington.edu Fri Feb 17 12:15:44 2006 3, 19 -- Received: from mxout5.cac.washington.edu (mxout5.cac.washington.edu [140.142.32.135]) 3, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HIFhWL007641 3, 19 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 12:15:44 -0600 3, 19 -- Received: from homer24.u.washington.edu (homer24.u.washington.edu [140.142.15.10]) 3, 19 -- by mxout5.cac.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k1HIFg5m011897 3, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 3, 19 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 10:15:43 -0800 3, 19 -- Received: from localhost (wpchan-at-localhost) 3, 19 -- by homer24.u.washington.edu (8.13.5+UW05.10/8.13.5+Submit) with ESMTP id k1HIFgup027346 3, 19 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 10:15:42 -0800 3, 19 -- Date: Fri, 17 Feb 2006 10:15:42 -0800 (PST) 3, 19 -- From: "W. Chan" {wpchan-at-u.washington.edu} 3, 19 -- To: microscopy-at-microscopy.com 3, 19 -- Subject: manual for Denton DV-502A 3, 19 -- Message-ID: {Pine.LNX.4.64.0602171008220.7757-at-homer24.u.washington.edu} 3, 19 -- MIME-Version: 1.0 3, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 3, 19 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_7 0' ==============================End of - Headers==============================
For our new IMAGE building, I specified PVC piping for the closed circ loop between the water chillers and scopes. We still notice a greenish sludge building up in the water filters (takes about 6-8 months to become significant) but I am certain that this is coming from the EM (copper cooling coils and iron connections---} electrolytic reaction). The EM service people told us that if we ever used acid to clean the lines that they would no longer warranty the microscope. The PVC lines are perfectly clean.
JB
} For years we have been wondering where the dark material was coming from } that accumulated in our water filters. These are filters in the closed } circuit lines between our microscopes and water recirculating units. The } lines are mainly in the ceiling or totally insulated as they run down the } walls in the scope rooms. Imaging our surprise when we went to do a minor } repair on one and watched as the plumber removed a regulator and inserted a } piece of galvanized pipe. } } Apparently when the building was built (20 years ago), the contractor } used galvanized pipe when copper had been specified. As it was hidden, we } did not know about the switch. All visible lines hooking up the chiller } compressor cooling with building water were copper. } } Well now these galvanized lines are really breaking down and clogging the } water pumps. We intend to replace all but were questioning whether it would } be best to replace with copper or PVC piping. Any suggestions? } } One concern was whether there would be the need to acid clean these lines in } the future (this is done routinely to the compressor lines to remove mineral } build-up). Since it is closed circuit, we should not accumulate large } amounts of minerals even though tap or deionized water will be used for the } system. We also can control algae growth with chemicals. Any suggestions on } this and should this dictate which material is used for the pipes? } } 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 Feb 17 11:51:22 2006 } 8, 21 -- Received: from exchange.purdue.edu } (1061exfe02.adpc.purdue.edu [128.210.63.223]) } 8, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k1HHpMgE030390 } 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 } 11:51:22 -0600 } 8, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by } exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 8, 21 -- Fri, 17 Feb 2006 12:51:21 -0500 } 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, 17 Feb 2006 17:51:21 +0000 } 8, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 } 8, 21 -- Date: Fri, 17 Feb 2006 12:51:20 -0500 } 8, 21 -- Subject: Microscope cooling lines } 8, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} } 8, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 8, 21 -- Message-ID: {C01B7748.D5AB%dsherman-at-purdue.edu} } 8, 21 -- Thread-Topic: Microscope cooling lines } 8, 21 -- Thread-Index: AcYz6sIxAPacp5/eEdqwBAARJN08Mg== } 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: 17 Feb 2006 17:51:21.0666 (UTC) } FILETIME=[C32F5220:01C633EA] } ==============================End of - Headers==============================
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 9, 18 -- From bozzola-at-siu.edu Fri Feb 17 12:57:04 2006 9, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HIv3LI017656 9, 18 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 12:57:04 -0600 9, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 9, 18 -- by abbmta2.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k1HIv252005395 9, 18 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 12:57:03 -0600 (CST) 9, 18 -- Mime-Version: 1.0 9, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 9, 18 -- Message-Id: {p0611040bc01bcb431cdd-at-[131.230.177.142]} 9, 18 -- In-Reply-To: {200602171754.k1HHsAVw032278-at-ns.microscopy.com} 9, 18 -- References: {200602171754.k1HHsAVw032278-at-ns.microscopy.com} 9, 18 -- Date: Fri, 17 Feb 2006 12:57:00 -0600 9, 18 -- To: Microscopy-at-msa.microscopy.com 9, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 9, 18 -- Subject: Re: [Microscopy] Microscope cooling lines 9, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 9, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
Another material you may wish to consider is called PEX, which is a cross-linked polyethylene. I don't know too much about its characteristics, except that it's very smooth inside, which should retard crud accumulation and it's more opaque than white pvc. It may be worth checking out.
Paul
-------------------------------------------------------------------------- Famous Last Words Department: "I did not get my Spaghetti-O's, I got spaghetti. I want the press to know this." ~~ Thomas J. Grasso, d. March 20, 1995 Executed by injection, Oklahoma.
==============================Original Headers============================== 7, 21 -- From pgrover-at-bilbo.bio.purdue.edu Fri Feb 17 13:19:13 2006 7, 21 -- Received: from mailhub128.itcs.purdue.edu (mailhub128.itcs.purdue.edu [128.210.5.128]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HJJCk8027344 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 13:19:12 -0600 7, 21 -- Received: from paklabpgrover (dhcp155-122.bio.purdue.edu [128.210.155.122]) 7, 21 -- by mailhub128.itcs.purdue.edu (8.13.4/8.13.4/internal-smtp) with ESMTP id k1HJJCLp010612 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 14:19:12 -0500 7, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} 7, 21 -- To: {microscopy-at-microscopy.com} 7, 21 -- Subject: re: microscope cooling lines 7, 21 -- Date: Fri, 17 Feb 2006 14:19:17 -0500 7, 21 -- Message-ID: {000301c633f7$0bb24910$7a9bd280-at-paklabpgrover} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- Thread-Index: AcYz9wt9iKTkG2SVRtGWvOlC3m8itw== 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 7, 21 -- X-PMX-Version: 4.7.1.128075 7, 21 -- X-PerlMx-Virus-Scanned: Yes ==============================End of - Headers==============================
It would be good to know what is the operating temperature, pressure, flow rate and the characteristics of the water feed for make-up.
There is no material in-compatibility between copper piping and PVC piping. They can be used in the same water curcuit.
If you want to know if there is a problem using copper, you can take some of the water out of the system and see how much copper is present. You need to have a base line of copper from the water source, so you should take a water sample from the source also. There is likely not a problem, it depends upon your water chemistry. Knowing your water chemistry is fundamental to knowing what piping is preferred.
You may have building code restrictions regarding PVC piping that is hidden, which may be the reason the orginal contractor put in galvanized piping. You will have to look at what codes apply to where you are.
Regarding acid cleaning, you should know what your deposits are in your piping before deciding how to clean them. As an FYI, copper will generally corrode at pH's below about 6.3 or so. There are low pH cleaners that can be used with copper, but they contain corrosion inhibitors.
dj
On Fri, 17 Feb 2006 dsherman-at-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 } ---------------------------------------------------------------------------- } } Listers, } } For years we have been wondering where the dark material was coming from } that accumulated in our water filters. These are filters in the closed } circuit lines between our microscopes and water recirculating units. The } lines are mainly in the ceiling or totally insulated as they run down the } walls in the scope rooms. Imaging our surprise when we went to do a minor } repair on one and watched as the plumber removed a regulator and inserted a } piece of galvanized pipe. } } Apparently when the building was built (20 years ago), the contractor } used galvanized pipe when copper had been specified. As it was hidden, we } did not know about the switch. All visible lines hooking up the chiller } compressor cooling with building water were copper. } } Well now these galvanized lines are really breaking down and clogging the } water pumps. We intend to replace all but were questioning whether it would } be best to replace with copper or PVC piping. Any suggestions? } } One concern was whether there would be the need to acid clean these lines in } the future (this is done routinely to the compressor lines to remove mineral } build-up). Since it is closed circuit, we should not accumulate large } amounts of minerals even though tap or deionized water will be used for the } system. We also can control algae growth with chemicals. Any suggestions on } this and should this dictate which material is used for the pipes? } } 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 Feb 17 11:51:22 2006 } 8, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) } 8, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HHpMgE030390 } 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 11:51:22 -0600 } 8, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 8, 21 -- Fri, 17 Feb 2006 12:51:21 -0500 } 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, 17 Feb 2006 17:51:21 +0000 } 8, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 } 8, 21 -- Date: Fri, 17 Feb 2006 12:51:20 -0500 } 8, 21 -- Subject: Microscope cooling lines } 8, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} } 8, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 8, 21 -- Message-ID: {C01B7748.D5AB%dsherman-at-purdue.edu} } 8, 21 -- Thread-Topic: Microscope cooling lines } 8, 21 -- Thread-Index: AcYz6sIxAPacp5/eEdqwBAARJN08Mg== } 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: 17 Feb 2006 17:51:21.0666 (UTC) FILETIME=[C32F5220:01C633EA] } ==============================End of - Headers============================== }
==============================Original Headers============================== 10, 25 -- From dljones-at-bestweb.net Fri Feb 17 13:24:26 2006 10, 25 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) 10, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HJOOlm003358 10, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 13:24:26 -0600 10, 25 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) 10, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id 747541CD06; 10, 25 -- Fri, 17 Feb 2006 14:24:23 -0500 (EST) 10, 25 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) 10, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id CAD7C1CCC2; 10, 25 -- Fri, 17 Feb 2006 14:24:21 -0500 (EST) 10, 25 -- Date: Fri, 17 Feb 2006 14:33:39 -0500 (Eastern Standard Time) 10, 25 -- From: "David L. Jones" {dljones-at-bestweb.net} 10, 25 -- To: dsherman-at-purdue.edu 10, 25 -- cc: Microscopy-at-microscopy.com 10, 25 -- Subject: Re: [Microscopy] Microscope cooling lines 10, 25 -- In-Reply-To: {200602171802.k1HI2DeL005131-at-ns.microscopy.com} 10, 25 -- Message-ID: {Pine.WNT.4.62.0602171405030.1080-at-dlj} 10, 25 -- References: {200602171802.k1HI2DeL005131-at-ns.microscopy.com} 10, 25 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 10, 25 -- MIME-Version: 1.0 10, 25 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 10, 25 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net 10, 25 -- X-Spam-Level: 10, 25 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed 10, 25 -- version=3.0.2 ==============================End of - Headers==============================
On Feb 17, 2006, at 9:51 AM, dsherman-at-purdue.edu wrote:
} For years we have been wondering where the dark material was coming } from } that accumulated in our water filters. These are filters in the closed } circuit lines between our microscopes and water recirculating units. } The } lines are mainly in the ceiling or totally insulated as they run down } the } walls in the scope rooms. Imaging our surprise when we went to do a } minor } repair on one and watched as the plumber removed a regulator and } inserted a } piece of galvanized pipe. } } Apparently when the building was built (20 years ago), the } contractor } used galvanized pipe when copper had been specified. As it was } hidden, we } did not know about the switch. All visible lines hooking up the } chiller } compressor cooling with building water were copper. } } Well now these galvanized lines are really breaking down and clogging } the } water pumps. We intend to replace all but were questioning whether it } would } be best to replace with copper or PVC piping. Any suggestions? } } One concern was whether there would be the need to acid clean these } lines in } the future (this is done routinely to the compressor lines to remove } mineral } build-up). Since it is closed circuit, we should not accumulate large } amounts of minerals even though tap or deionized water will be used } for the } system. We also can control algae growth with chemicals. Any } suggestions on } this and should this dictate which material is used for the pipes? } Dear Debby, If you intend to clean the lines with acid, I suggest PVC, since Cu can be etched at low pH. In addition to floating some dichlorophene for algal control, we add a corrosion inhibitor. We have been using a Mo-based formula, which was available from Aqua Labs on the East coast and from Skasol on the West coast, so find a distributor in your area. I think that either Aqua or Skasol would be able to give you that info. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Fri Feb 17 13:36:46 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HJakQG013933 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 13:36:46 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 040B535C5A 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 11:36:46 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id AC201340D7 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 11:36:43 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602171751.k1HHpVq9030559-at-ns.microscopy.com} 4, 22 -- References: {200602171751.k1HHpVq9030559-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {6f18d2977f3ac75cf6e6159ba16304e9-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Microscope cooling lines 4, 22 -- Date: Fri, 17 Feb 2006 11:44:38 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
On Feb 17, 2006, at 11:19 AM, pgrover-at-bilbo.bio.purdue.edu wrote:
} Another material you may wish to consider is called PEX, which is a } cross-linked polyethylene. I don't know too much about its } characteristics, } except that it's very smooth inside, which should retard crud } accumulation } and it's more opaque than white pvc. It may be worth checking out. } Dear Paul, PEX sounds like a better material than PVC. It should be essentially inert--I expect that it will withstand concentrated bases and acids and would be unaffected by organic solvents (not that one would want to run those through the scope lines). Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Fri Feb 17 13:42:57 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HJgvr6022901 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 13:42:57 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 28C8235764 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 11:42:57 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id 4558E340D7 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Feb 2006 11:42:56 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602171919.k1HJJIaZ027514-at-ns.microscopy.com} 4, 22 -- References: {200602171919.k1HJJIaZ027514-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {929b43e7c910fd8036e0032d59e72528-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] re: microscope cooling lines 4, 22 -- Date: Fri, 17 Feb 2006 11:50:51 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
As an FYI, if you have copper based and iron based materials mixed in the same system, the iron will corrode preferentially through galvanic coupling, not the copper. In fact, the iron becomes a sacrifical anode protecting the copper from corrosion. Any time you have those two materials in the same system, you should have dielectric couplings between the two or you will actively corrode the ferrous based material.
If you are having a copper corrosion problem, you should be looking elsewhere for the cause...
dj
On Fri, 17 Feb 2006 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 } ---------------------------------------------------------------------------- } } Debby, } } For our new IMAGE building, I specified PVC piping for the closed } circ loop between the water chillers and scopes. We still notice a } greenish sludge building up in the water filters (takes about 6-8 } months to become significant) but I am certain that this is coming } from the EM (copper cooling coils and iron } connections---} electrolytic reaction). The EM service people told us } that if we ever used acid to clean the lines that they would no } longer warranty the microscope. The PVC lines are perfectly clean. } } JB }
==============================Original Headers============================== 7, 25 -- From dljones-at-bestweb.net Fri Feb 17 13:43:51 2006 7, 25 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) 7, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HJhoE0024591 7, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 13:43:51 -0600 7, 25 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) 7, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id EDFB81CD04; 7, 25 -- Fri, 17 Feb 2006 14:43:49 -0500 (EST) 7, 25 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) 7, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id 7E4831CCC2; 7, 25 -- Fri, 17 Feb 2006 14:43:48 -0500 (EST) 7, 25 -- Date: Fri, 17 Feb 2006 14:53:06 -0500 (Eastern Standard Time) 7, 25 -- From: "David L. Jones" {dljones-at-bestweb.net} 7, 25 -- To: bozzola-at-siu.edu 7, 25 -- cc: Microscopy-at-microscopy.com 7, 25 -- Subject: Re: [Microscopy] Re: Microscope cooling lines 7, 25 -- In-Reply-To: {200602171902.k1HJ2UNa024787-at-ns.microscopy.com} 7, 25 -- Message-ID: {Pine.WNT.4.62.0602171446420.1080-at-dlj} 7, 25 -- References: {200602171902.k1HJ2UNa024787-at-ns.microscopy.com} 7, 25 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 7, 25 -- MIME-Version: 1.0 7, 25 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 7, 25 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net 7, 25 -- X-Spam-Level: 7, 25 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed 7, 25 -- version=3.0.2 ==============================End of - Headers==============================
Along these lines, I noticed a lot more copper leaching into the lines when I used deionized water than regular or filtered tap water.
Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze intended for cars should have corrosion inhibitors and should be suitable for this purpose.
I have still gotten green particles regardless of whether antifreeze was used or not and have regularly made it a practice to clean the filters in the lines once a semester. Jerry Calvin
At 1:46 PM -0600 2/17/06, dljones-at-bestweb.net wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- **************************** Jerry G. Calvin Science Support Technician Box 0731 Biology Department Vassar College 124 Raymond Avenue Poughkeepsie, NY 12604-0731
} PEX sounds like a better material than PVC. It should be essentially
Beware that PEX will degrade over time in sunlight (ultraviolet). That said, I have PEX in my house instead of Cu, works well and is easy to run, however all transitions through the walls are Cu fittings so the PEX stays in the dark.
Scott -- ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
==============================Original Headers============================== 4, 16 -- From davilla-at-4pi.com Fri Feb 17 15:17:27 2006 4, 16 -- Received: from mx.4pi.com (mx.4pi.com [24.172.19.59]) 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1HLHQjh021101 4, 16 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 15:17:27 -0600 4, 16 -- Received: from [24.172.19.62] (HELO [192.168.9.10]) 4, 16 -- by mx.4pi.com (Stalker SMTP Server 1.8b9d14) 4, 16 -- with ESMTP id S.0000850373 for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:17:26 -0500 4, 16 -- Mime-Version: 1.0 4, 16 -- Message-Id: {p06230957c01bece6d048-at-[192.168.9.10]} 4, 16 -- In-Reply-To: {200602171943.k1HJh1eC023022-at-ns.microscopy.com} 4, 16 -- References: {200602171943.k1HJh1eC023022-at-ns.microscopy.com} 4, 16 -- Date: Fri, 17 Feb 2006 16:17:38 -0500 4, 16 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 4, 16 -- From: "Scott D. Davilla" {davilla-at-4pi.com} 4, 16 -- Subject: Re: [Microscopy] microscope cooling lines 4, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Yes, it is not unusual to find more copper leaching into the lines with deionized water than tap water. It does depend upon the tap water chemistry.
I would not recommend antifreeze for cars, however, I would recommend to use HVAC grade propylene glycol. That would include inhibitors to protect the metals in the system and it includes various stablizers for the glycol. I guess if there are glycols for cars that are propylene glycol based with inhibitors, those may be OK. I most certainly would not use ethylene glycol based antifreeze.
I would ask you what kind of antifreeze you are using and have you done an EDS spectrum of your green deposit from you filters? I would think further discussion of your specifics may best be addressed off the list...
dj
On Fri, 17 Feb 2006, Jerry Calvin wrote:
} Along these lines, I noticed a lot more copper leaching into the lines when } I used deionized water than regular or filtered tap water. } } Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze intended } for cars should have corrosion inhibitors and should be suitable for this } purpose. } } I have still gotten green particles regardless of whether antifreeze was used } or not and have regularly made it a practice to clean the filters in the } lines once a semester. Jerry Calvin } } } } At 1:46 PM -0600 2/17/06, dljones-at-bestweb.net wrote: } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } JB, } } } } As an FYI, if you have copper based and iron based materials mixed in the } } same } } system, the iron will corrode preferentially through galvanic coupling, not } } the } } copper. In fact, the iron becomes a sacrifical anode protecting the copper } } from } } corrosion. Any time you have those two materials in the same system, you } } should } } have dielectric couplings between the two or you will actively corrode the } } ferrous based material. } } } } If you are having a copper corrosion problem, you should be looking } } elsewhere } } for the cause... } } } } dj } } } } On Fri, 17 Feb 2006 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 } } } ---------------------------------------------------------------------------- } } } } } } Debby, } } } } } } For our new IMAGE building, I specified PVC piping for the closed } } } circ loop between the water chillers and scopes. We still notice a } } } greenish sludge building up in the water filters (takes about 6-8 } } } months to become significant) but I am certain that this is coming } } } from the EM (copper cooling coils and iron } } } connections---} electrolytic reaction). The EM service people told us } } } that if we ever used acid to clean the lines that they would no } } } longer warranty the microscope. The PVC lines are perfectly clean. } } } } } } JB } } } } } } } } } ==============================Original } } Headers============================== } } 7, 25 -- From dljones-at-bestweb.net Fri Feb 17 13:43:51 2006 } } 7, 25 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net } } [209.94.103.46]) } } 7, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } k1HJhoE0024591 } } 7, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 13:43:51 } } -0600 } } 7, 25 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) } } 7, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id EDFB81CD04; } } 7, 25 -- Fri, 17 Feb 2006 14:43:49 -0500 (EST) } } 7, 25 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net } } [71.249.9.96]) } } 7, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id 7E4831CCC2; } } 7, 25 -- Fri, 17 Feb 2006 14:43:48 -0500 (EST) } } 7, 25 -- Date: Fri, 17 Feb 2006 14:53:06 -0500 (Eastern Standard Time) } } 7, 25 -- From: "David L. Jones" {dljones-at-bestweb.net} } } 7, 25 -- To: bozzola-at-siu.edu } } 7, 25 -- cc: Microscopy-at-microscopy.com } } 7, 25 -- Subject: Re: [Microscopy] Re: Microscope cooling lines } } 7, 25 -- In-Reply-To: {200602171902.k1HJ2UNa024787-at-ns.microscopy.com} } } 7, 25 -- Message-ID: {Pine.WNT.4.62.0602171446420.1080-at-dlj} } } 7, 25 -- References: {200602171902.k1HJ2UNa024787-at-ns.microscopy.com} } } 7, 25 -- X-X-Sender: dljones-at-pop-croton.bestweb.net } } 7, 25 -- MIME-Version: 1.0 } } 7, 25 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed } } 7, 25 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on } } smtp2-1.bestweb.net } } 7, 25 -- X-Spam-Level: } } 7, 25 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED } } autolearn=failed } } 7, 25 -- version=3.0.2 } } ==============================End of - } } Headers============================== } } } -- } **************************** } Jerry G. Calvin } Science Support Technician } Box 0731 Biology Department } Vassar College } 124 Raymond Avenue } Poughkeepsie, NY 12604-0731 } } (845) 437-7423 - Office } (845) 437-7424 - Confocal Room } FAX: (845) 437-7315 } E-Mail: jecalvin-at-vassar.edu }
==============================Original Headers============================== 8, 26 -- From dljones-at-bestweb.net Fri Feb 17 15:21:32 2006 8, 26 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) 8, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HLLVkH026422 8, 26 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 15:21:31 -0600 8, 26 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) 8, 26 -- by smtp2.bestweb.net (Postfix) with ESMTP id C09831CCF1; 8, 26 -- Fri, 17 Feb 2006 16:21:30 -0500 (EST) 8, 26 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) 8, 26 -- by smtp2.bestweb.net (Postfix) with ESMTP id 27CFB1CCE0; 8, 26 -- Fri, 17 Feb 2006 16:21:30 -0500 (EST) 8, 26 -- Date: Fri, 17 Feb 2006 16:30:47 -0500 (Eastern Standard Time) 8, 26 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 26 -- To: Jerry Calvin {jecalvin-at-vassar.edu} 8, 26 -- cc: Microscopy-at-microscopy.com 8, 26 -- Subject: Re: [Microscopy] Microscope cooling lines 8, 26 -- In-Reply-To: {a06001203c01bde1ab81a-at-[143.229.41.193]} 8, 26 -- Message-ID: {Pine.WNT.4.62.0602171608170.1080-at-dlj} 8, 26 -- References: {200602171946.k1HJkq7E000347-at-ns.microscopy.com} 8, 26 -- {a06001203c01bde1ab81a-at-[143.229.41.193]} 8, 26 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 26 -- MIME-Version: 1.0 8, 26 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 26 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net 8, 26 -- X-Spam-Level: 8, 26 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed 8, 26 -- version=3.0.2 ==============================End of - Headers==============================
A colleague asked me why she is so much more aware of the floaters in her eyes when she is using the microscope (or the telescope) than at other times. I assured her that it wasn't just her, it happens to a lot of us. But why floaters are so much more noticeable when looking in the microscope I do not know? Any suggestions?
Bob -- C. Robert Bagnell, Jr., Ph.D. Professor and Director, Microscopy Services Laboratory Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill Chapel Hill, NC 27599 phone 919-966-2413 fax 919-966-6718 e-mail bagnell-at-med.unc.edu web http://www.med.unc.edu/microscopy
==============================Original Headers============================== 2, 21 -- From bagnell-at-med.unc.edu Fri Feb 17 16:01:56 2006 2, 21 -- Received: from smtp-mx.med.unc.edu (fletcher.med.unc.edu [152.19.4.46]) 2, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HM1u7m008604 2, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:01:56 -0600 2, 21 -- Received: from castlehayne.med.unc.edu (castlehayne.med.unc.edu [152.19.4.29]) 2, 21 -- by smtp-mx.med.unc.edu (8.13.4+Sun/8.13.4) with ESMTP id k1HM1qc1025249 2, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 17:01:56 -0500 (EST) 2, 21 -- Received: from [152.19.80.19] by msrv0.med.unc.edu 2, 21 -- (Sun Java System Messaging Server 6.2-5.02 (built Dec 1 2005)) 2, 21 -- with ESMTPA id {0IUU00C9DQJ236B0-at-msrv0.med.unc.edu} for 2, 21 -- microscopy-at-microscopy.com; Fri, 17 Feb 2006 17:01:51 -0500 (EST) 2, 21 -- Date: Fri, 17 Feb 2006 17:01:49 -0500 2, 21 -- From: Robert Bagnell {bagnell-at-med.unc.edu} 2, 21 -- Subject: Floaters 2, 21 -- X-Sender: rml-at-imap-ns.med.unc.edu 2, 21 -- To: microscopy-at-microscopy.com 2, 21 -- Message-id: {p06110400c01bdabc3b81-at-[152.19.80.19]} 2, 21 -- MIME-version: 1.0 2, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 2, 21 -- Content-transfer-encoding: 7BIT 2, 21 -- X-Scanned-By: UNC/SoM/OIS/Mail_Filter on 152.19.4.46 ==============================End of - Headers==============================
My service engineer recommends neither tap nor distilled water, but rather bottled spring water. Has anyone yet mentioned the possibility that green sludge in the filter might be algae growing in the cooling water or the cooling unit? --Jan Factor
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College, State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
==============================Original Headers============================== 4, 16 -- From jfactor-at-ns.purchase.edu Fri Feb 17 16:02:30 2006 4, 16 -- Received: from zephyr.ns.purchase.edu (zephyr.ns.purchase.edu [199.79.168.193]) 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HM2PAx009529 4, 16 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:02:25 -0600 4, 16 -- Received: from [10.52.0.79] ([10.52.0.79]) 4, 16 -- by zephyr.ns.purchase.edu (AIX5.1/8.11.6p2/8.11.0) with ESMTP id k1HM5lr17594; 4, 16 -- Fri, 17 Feb 2006 17:05:48 -0500 4, 16 -- Message-ID: {43F647F0.1070409-at-ns.purchase.edu} 4, 16 -- Date: Fri, 17 Feb 2006 17:02:24 -0500 4, 16 -- From: Jan Factor {jfactor-at-ns.purchase.edu} 4, 16 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 4, 16 -- MIME-Version: 1.0 4, 16 -- To: microscopy-at-microscopy.com 4, 16 -- Subject: Re: Microscope cooling lines 4, 16 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 16 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Question: Can anybody provide an explanation concerning the refractive index -dependency of the interference colours seen in charts for estimating the thickness of ultrathin sections? The small print in such charts usually reads "valid for refractive index of ca. 1.5", which is fine for methacrylates and similar embedding media, but what type of shift (if significant) would be observed for a lower refractive index, say 1.3? I am not sure whether the usual charts are valid for the interference colours given by cryosections.
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Title-Subject: [Filtered] NTA-nanogold for His tagged protein in EM?
Question: Has anynone had success using NTA-nanogold to disclose the location of His-tagged proteins by electron microscopy? I would like to attach the nanogold this way before injecting the protein into a cell. An alternative would be to apply gold labelled primary anti-His after fixation and permeabilization.
4 Student Scholarships ($500) Available for support for attending and presenting a paper/poster
PARTICLE WORKSHOP 2006
A Joint NIST / Microbeam Analysis Society Special Topics Workshop
Dates: April 24th - 26th 2006
Location: NIST, Gaithersburg, MD
The analysis of microscopic particles represents a measurement challenge in a diverse range of scientific, technological and environmental disciplines. Examples of areas of interest include nanoscale particles as building blocks for nanotechnology, contamination in high technology manufacturing, trace forensic detection for homeland security, and environmental sampling and monitoring. Characterization of particle specimens is complicated by mass, volume and morphological effects that cause well established bulk techniques to break down. In addition, while the measurement types under consideration at this workshop are performed on a particle-by-particle basis, the properties of interest are often statistical measures of populations of similar particles.
This workshop will focus on the challenges that face industrial and governmental application of microscopic particle measurement techniques. The practical tone of the meeting will set by speakers representing various industries and governmental organizations who will provide insight into their particle measurement requirements. These speakers will be followed by sessions that focus on sample preparation and statistical experimental design and established & emerging measurement techniques. Instrumentation to be discussed include SEM/EDS, AEM, TOF/SIMS, optical, FIB and scanned probe. Each of these sessions will be concluded by a round table discussion in which attendees are encouraged to contribute.
Registration Deadline: March 31, 2006
Students interested in applying for one of the 4 student scholarships should immediately contact
Nicholas W. M. Ritchie NIST Microanalysis Research Group 100 Bureau Drive STOP 8371 Gaithersburg, MD 20899-8371 301-975-3929 nicholas.ritchie-at-nist.gov
There is no registration fee: the workshop is free to all attendees. However, space is limited and pre-registration is required by NIST Security for entry onto the NIST campus. This rule is strictly enforced.
Full information at www.cstl.nist.gov/div837/Division/meetings/particleworkshop/particle.htm
==============================Original Headers============================== 9, 24 -- From johnf-at-geology.wisc.edu Fri Feb 17 16:21:54 2006 9, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 9, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HMLs1o013872 9, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 17 Feb 2006 16:21:54 -0600 9, 24 -- Received: from localhost (localhost [127.0.0.1]) 9, 24 -- by localhost (Postfix) with ESMTP id F0BEE20D01 9, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 17 Feb 2006 16:21:53 -0600 (CST) 9, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 9, 24 -- by localhost (ice [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 9, 24 -- id 07362-03 for {Microscopy-at-Microscopy.Com} ; 9, 24 -- Fri, 17 Feb 2006 16:21:49 -0600 (CST) 9, 24 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 9, 24 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 9, 24 -- (No client certificate requested) 9, 24 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 9E1EA20D16 9, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 17 Feb 2006 16:21:49 -0600 (CST) 9, 24 -- Mime-Version: 1.0 9, 24 -- Message-Id: {p0623091dc01bfc64a0eb-at-[144.92.206.57]} 9, 24 -- Date: Fri, 17 Feb 2006 16:19:32 -0600 9, 24 -- To: Microscopy-at-Microscopy.Com 9, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 9, 24 -- Subject: student scholarships: NIST/MAS Particle Workshop April 2006 9, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 9, 24 -- X-Virus-Scanned: by amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
It has to do with the physics of simple pinhole optics. Essentially, when you are just in the right focal plane, you are doing an "entopic exam" of your eye. You can also reproduce the experiment by putting a small hole (about 1/8") into a piece of cardboard (1/2 of a file folder) and staring through it at a neutral surface (blank wall; sky, if not too bright). Adjust the distance between the card and your eye and, at the right distance, you will see the internal structure.
I've had an interesting array of tiny cataracts for over 25 years and keep track of their position and size using this method.
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
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 04:05 PM 2/17/2006, bagnell-at-med.unc.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 15, 17 -- From bfoster-at-mme1.com Fri Feb 17 16:31:50 2006 15, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 15, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HMVnJe023336 15, 17 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:31:49 -0600 15, 17 -- Received: (qmail 19789 invoked from network); 17 Feb 2006 17:33:30 -0500 15, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 15, 17 -- by enterprise.5starpro.com with SMTP; 17 Feb 2006 17:33:30 -0500 15, 17 -- Message-Id: {7.0.1.0.0.20060217162833.01fc9ec0-at-mme1.com} 15, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 17 -- Date: Fri, 17 Feb 2006 16:31:54 -0600 15, 17 -- To: bagnell-at-med.unc.edu, Microscopy Listserver {microscopy-at-microscopy.com} 15, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 15, 17 -- Subject: Re: [Microscopy] Floaters 15, 17 -- In-Reply-To: {200602172205.k1HM5BbH017110-at-ns.microscopy.com} 15, 17 -- References: {200602172205.k1HM5BbH017110-at-ns.microscopy.com} 15, 17 -- Mime-Version: 1.0 15, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I had the same sludge problem about 1.5 years ago in a new Haskris chiller. Haskris recommends using only distilled water. That lasted about three months and the slime appeared. It was a combination of algae and small particles. Dumped the water and replaced with new distilled water and Skasol to flush. Then, new distilled water and one half liter of Hexid A4 from Applied Thermal Control Ltd. UK as supplied by SEM service tech. Chiller seized after about three months. Post mortem indicated that the impeller blades failed. Most likely due to misalignment of motor and pump.
Haskris replaced motor and pump assembly. Fluid was drained and replaced with distilled water and ethylene glycol (.5G to 4.5G distilled water). Filters were changed and no problems for about eight months. Liquid is not starting to become darker and small build up of stuff in chiller main filter. It is time to change liquid and filters. Main filter is in the water tank and is spec'd at about 50u. External toilet paper style filter is spec'd at 2u. So both get changed at the same time.
The Haskris unit specifically says to not use automotive antifreeze since it will deteriorate the BUNA N material in the chiller. Some anti freeze contains ethylene glycol. So I'm puzzled by the successful use of distilled water and EG. Perhaps they meant to say not to use 100% antifreeze rather than a diluted mix.
The other factor is that the SEM came with basically transparent water hoses. This is not good since the light gets into them and advances the algae. So this upcoming liquid and filter replacement will include replacing the hoses with opaque ones.
Overall, there are three aspects to be concerned about:
1. chiller guts and pump 2. hoses 3. SEM items that get chilled water (TMP, coils, etc.)
I don't think that there is a single simple answer to this problem since SEMs are different and chillers are different.
gary g.
At 02:04 PM 2/17/2006, you wrote:
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==============================Original Headers============================== 15, 20 -- From gary-at-gaugler.com Fri Feb 17 16:58:51 2006 15, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 15, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1HMwoD3002918 15, 20 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:58:51 -0600 15, 20 -- Received: (qmail 24885 invoked from network); 17 Feb 2006 14:58:50 -0800 15, 20 -- Received: by simscan 1.1.0 ppid: 24881, pid: 24883, t: 0.0890s 15, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 15, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 15, 20 -- by qsmtp3 with SMTP; 17 Feb 2006 14:58:50 -0800 15, 20 -- Message-Id: {6.2.3.4.2.20060217143542.0205ed40-at-mail.calweb.com} 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 15, 20 -- Date: Fri, 17 Feb 2006 14:58:51 -0800 15, 20 -- To: jfactor-at-ns.purchase.edu 15, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 15, 20 -- Subject: Re: [Microscopy] Re: Microscope cooling lines 15, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 15, 20 -- In-Reply-To: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} 15, 20 -- References: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} 15, 20 -- Mime-Version: 1.0 15, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Just a couple of points I'd like to make w.r.t. your post.
I would not recommend using ethylene glycol as it is toxic. Propylene glycol is non-toxic.
The problem with using "antifreeze" mixtures not intended for use in these kinds of systems is that they do not usually contain the proper additives. Glycol solutions that are not HVAC grade will deteriorate over time through a type of polymerization that will plug things up and render the system inoperable. The resulting deposits thus formed are very inert and to my knowledge no one has ever found a way to clean them so you basically have to replace the chiller system.
HVAC grade polypropylene contains additives to avoid that problem plus inhibitors that stop corrosion of most commercially available materials in piping. That also includes seal materials, but I'm not sure about specifically N-buena seals. I'd have to look that up, but I would think it also compatible being such a commonly used seal material.
Biofouling is quite common in closed loop systems. Using a biocide is usually used in these systems to eliminate this problem.
The original poster to this thread likely is in a location where they have a professional water treatment company taking care of large HVAC systems. Perhaps they should talk with the representative of that company and find out what is being done for chemical treatment of chilled water systems there. They may be able to just get some of the proper chemicals that likely exist on-site already.
I would also like to point out, there is really no reason to go through the expense of using a glycol based system unless there is danger of freezing the coolant for some reason. There are numerous other water treatments that are much less expensive and work very well to keep a closed loop system running well. If there is little to no make up water needed for the closed system, once set-up properly, there is little more to do other than enjoy a clean running system...
dj
On Fri, 17 Feb 2006, gary-at-gaugler.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I had the same sludge problem about 1.5 years ago in } a new Haskris chiller. Haskris recommends using only } distilled water. That lasted about three months and } the slime appeared. It was a combination of algae and } small particles. Dumped the water and replaced with } new distilled water and Skasol to flush. Then, new } distilled water and one half liter of Hexid A4 from } Applied Thermal Control Ltd. UK as supplied by SEM } service tech. Chiller seized after about three months. } Post mortem indicated that the impeller blades failed. } Most likely due to misalignment of motor and pump. } } Haskris replaced motor and pump assembly. Fluid was } drained and replaced with distilled water and ethylene } glycol (.5G to 4.5G distilled water). Filters were } changed and no problems for about eight months. Liquid } is not starting to become darker and small build up of } stuff in chiller main filter. It is time to change liquid and } filters. Main filter is in the water tank and is spec'd } at about 50u. External toilet paper style filter is spec'd } at 2u. So both get changed at the same time. } } The Haskris unit specifically says to not use automotive } antifreeze since it will deteriorate the BUNA N material in } the chiller. Some anti freeze contains ethylene glycol. } So I'm puzzled by the successful use of distilled water and } EG. Perhaps they meant to say not to use 100% antifreeze } rather than a diluted mix. } } The other factor is that the SEM came with basically transparent } water hoses. This is not good since the light gets into them } and advances the algae. So this upcoming liquid and filter } replacement will include replacing the hoses with opaque ones. } } Overall, there are three aspects to be concerned about: } } 1. chiller guts and pump } 2. hoses } 3. SEM items that get chilled water (TMP, coils, etc.) } } I don't think that there is a single simple answer to this } problem since SEMs are different and chillers are different. } } gary g. } } } } At 02:04 PM 2/17/2006, you wrote: } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } My service engineer recommends neither tap nor distilled water, but } } rather bottled spring water. Has anyone yet mentioned the possibility } } that green sludge in the filter might be algae growing in the cooling } } water or the cooling unit? } } --Jan Factor } } } } --------------------------------------- } } Jan Robert Factor, Ph.D. } } Professor of Biology } } --------------------------------------- } } Natural Sciences } } Purchase College, State University of New York } } 735 Anderson Hill Rd. } } Purchase, NY 10577 } } USA } } --------------------------------------- } } Office Tel: 914-251-6659 } } Office Fax: 914-251-6635 } } E-mail: jfactor-at-ns.purchase.edu } } or- jan.factor-at-purchase.edu } } --------------------------------------- } } } } } } } } ==============================Original Headers============================== } } 4, 16 -- From jfactor-at-ns.purchase.edu Fri Feb 17 16:02:30 2006 } } 4, 16 -- Received: from zephyr.ns.purchase.edu } } (zephyr.ns.purchase.edu [199.79.168.193]) } } 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } k1HM2PAx009529 } } 4, 16 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 } } 16:02:25 -0600 } } 4, 16 -- Received: from [10.52.0.79] ([10.52.0.79]) } } 4, 16 -- by zephyr.ns.purchase.edu (AIX5.1/8.11.6p2/8.11.0) } } with ESMTP id k1HM5lr17594; } } 4, 16 -- Fri, 17 Feb 2006 17:05:48 -0500 } } 4, 16 -- Message-ID: {43F647F0.1070409-at-ns.purchase.edu} } } 4, 16 -- Date: Fri, 17 Feb 2006 17:02:24 -0500 } } 4, 16 -- From: Jan Factor {jfactor-at-ns.purchase.edu} } } 4, 16 -- User-Agent: Thunderbird 1.5 (Windows/20051201) } } 4, 16 -- MIME-Version: 1.0 } } 4, 16 -- To: microscopy-at-microscopy.com } } 4, 16 -- Subject: Re: Microscope cooling lines } } 4, 16 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } 4, 16 -- Content-Transfer-Encoding: 7bit } } ==============================End of - Headers============================== } } } ==============================Original Headers============================== } 15, 20 -- From gary-at-gaugler.com Fri Feb 17 16:58:51 2006 } 15, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) } 15, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1HMwoD3002918 } 15, 20 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:58:51 -0600 } 15, 20 -- Received: (qmail 24885 invoked from network); 17 Feb 2006 14:58:50 -0800 } 15, 20 -- Received: by simscan 1.1.0 ppid: 24881, pid: 24883, t: 0.0890s } 15, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 } 15, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } 15, 20 -- by qsmtp3 with SMTP; 17 Feb 2006 14:58:50 -0800 } 15, 20 -- Message-Id: {6.2.3.4.2.20060217143542.0205ed40-at-mail.calweb.com} } 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 } 15, 20 -- Date: Fri, 17 Feb 2006 14:58:51 -0800 } 15, 20 -- To: jfactor-at-ns.purchase.edu } 15, 20 -- From: Gary Gaugler {gary-at-gaugler.com} } 15, 20 -- Subject: Re: [Microscopy] Re: Microscope cooling lines } 15, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} } 15, 20 -- In-Reply-To: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} } 15, 20 -- References: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} } 15, 20 -- Mime-Version: 1.0 } 15, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } ==============================End of - Headers============================== }
==============================Original Headers============================== 12, 21 -- From "dljones-at-bestweb.net"-at-bestweb.net Fri Feb 17 20:43:13 2006 12, 21 -- Received: from mta4.srv.hcvlny.cv.net (mta4.srv.hcvlny.cv.net [167.206.4.199]) 12, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1I2hDlf008914 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 20:43:13 -0600 12, 21 -- Received: from localhost (ool-44c62883.dyn.optonline.net [68.198.40.131]) 12, 21 -- by mta4.srv.hcvlny.cv.net 12, 21 -- (Sun Java System Messaging Server 6.2-4.03 (built Sep 22 2005)) 12, 21 -- with ESMTP id {0IUV00D2N3JYUD00-at-mta4.srv.hcvlny.cv.net} for 12, 21 -- microscopy-at-microscopy.com; Fri, 17 Feb 2006 21:43:13 -0500 (EST) 12, 21 -- Date: Fri, 17 Feb 2006 21:43:12 -0500 (Eastern Standard Time) 12, 21 -- From: "David L. Jones" {"dljones-at-bestweb.net"-at-bestweb.net} 12, 21 -- Subject: Re: [Microscopy] Microscope cooling lines 12, 21 -- In-reply-to: {200602172305.k1HN5bhk012207-at-ns.microscopy.com} 12, 21 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 12, 21 -- To: gary-at-gaugler.com 12, 21 -- Cc: microscopy-at-microscopy.com 12, 21 -- Message-id: {Pine.WNT.4.64.0602172109370.408-at-dljtoshiba} 12, 21 -- MIME-version: 1.0 12, 21 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 12, 21 -- Content-transfer-encoding: 7BIT 12, 21 -- References: {200602172305.k1HN5bhk012207-at-ns.microscopy.com} ==============================End of - Headers==============================
If EG is toxic and PG is not, that is a factor. However, I do not care if one or the other is toxic. I do not swim in the 5 gallons of fluid. So, between the two glycols, which is best for SEM? I do not know. What do you think?
So there are toxic issues, corrosive issues, etc., etc. Well, then just dump the SEM, eh? No. I do not think so. One must work with the materials at hand. So, what do you think are the best materials for chiller fluid?
gary g.
At 06:43 PM 2/17/2006, you wrote: } Gary, } } Just a couple of points I'd like to make w.r.t. your post. } } I would not recommend using ethylene glycol as it is toxic. } Propylene glycol is non-toxic. } } The problem with using "antifreeze" mixtures not intended for use in } these kinds of systems is that they do not usually contain the } proper additives. Glycol solutions that are not HVAC grade will } deteriorate over time through a type of polymerization that will } plug things up and render the system inoperable. The resulting } deposits thus formed are very inert and to my knowledge no one has } ever found a way to clean them so you basically have to replace the } chiller system. } } HVAC grade polypropylene contains additives to avoid that problem } plus inhibitors that stop corrosion of most commercially available } materials in piping. That also includes seal materials, but I'm not } sure about specifically N-buena seals. I'd have to look that up, but } I would think it also compatible being such a commonly used seal material. } } Biofouling is quite common in closed loop systems. Using a biocide } is usually used in these systems to eliminate this problem. } } The original poster to this thread likely is in a location where } they have a professional water treatment company taking care of } large HVAC systems. Perhaps they should talk with the representative } of that company and find out what is being done for chemical } treatment of chilled water systems there. They may be able to just } get some of the proper chemicals that likely exist on-site already. } } I would also like to point out, there is really no reason to go } through the expense of using a glycol based system unless there is } danger of freezing the coolant for some reason. There are numerous } other water treatments that are much less expensive and work very } well to keep a closed loop system running well. If there is little } to no make up water needed for the closed system, once set-up } properly, there is little more to do other than enjoy a clean running system... } } dj } } On Fri, 17 Feb 2006, gary-at-gaugler.com wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 9, 21 -- From gary-at-gaugler.com Fri Feb 17 20:51:47 2006 9, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1I2pkQd018305 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 20:51:47 -0600 9, 21 -- Received: (qmail 22693 invoked from network); 17 Feb 2006 18:51:46 -0800 9, 21 -- Received: by simscan 1.1.0 ppid: 22690, pid: 22691, t: 0.2511s 9, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 21 -- by qsmtp3 with SMTP; 17 Feb 2006 18:51:46 -0800 9, 21 -- Message-Id: {6.2.3.4.2.20060217184632.02068418-at-mail.calweb.com} 9, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 9, 21 -- Date: Fri, 17 Feb 2006 18:51:46 -0800 9, 21 -- To: "David L. Jones" {"dljones-at-bestweb.net"-at-bestweb.net} 9, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 21 -- Subject: Re: [Microscopy] Microscope cooling lines 9, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 21 -- In-Reply-To: {Pine.WNT.4.64.0602172109370.408-at-dljtoshiba} 9, 21 -- References: {200602172305.k1HN5bhk012207-at-ns.microscopy.com} 9, 21 -- {Pine.WNT.4.64.0602172109370.408-at-dljtoshiba} 9, 21 -- Mime-Version: 1.0 9, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This is a perennial problem, I know, but does anyone have any effective "pet" solutions to the problem of charging and subsequent electrostatic arcing of TEM film in the extremely dry air we get in New England typically in the winter, and some other people get at other seasons of the year?
The discharges can occur at almost any stage of the handling, from getting the film out of the vendor's packing, loading and unloading in the microscope carriers and loading in the developing racks (we use Lucite ones). We have considered a humidifier in the darkroom, but that would involve blocking off the ventilation (we have make-up air positively blown into to room and exhaust actively sucked out) to prevent our moisture from being simply blown away.
Any hints would be welcome, but switching to digital imaging is one that is not going to happen.
Tony.
************************************* Anthony J. Garratt-Reed M.A. D.Phil. MIT Room #13-1027 77 Massachusetts Avenue Cambridge, Massachusetts 02139-4307 USA
Below are responses (abbreviated) to my question about which to use, copper or PVC, to replace water lines running from cooling units to microscopes. I will run any suggestions by my service engineers just to make sure there are no concerns regarding specific equipment. I do know that water pH is important and that may play a role in determining what additives to use:
I am a service engineer who works on electron microscopes for the last 17 years and my point of view: I would go with copper this way no matter what may or may not happen to the lines (cleaning out - whatever) you have no worries. (Ray Spengler)
I do not think that there will be a problem using copper or PVC. But you chould check with the eq maker to see if they have any issues. We use Copper and have not had any problems. We do get build up over time due to the hoses "rotting" over time and general scale buildup. (Karl Weiss)
USE PVC and make the final connection to the equipment with two coils of rubber hose for vibration control. ( Fran Laabs)
For our new IMAGE building, I specified PVC piping for the closed circ loop between the water chillers and scopes. We still notice a greenish sludge building up in the water filters (takes about 6-8 months to become significant) but I am certain that this is coming from the EM (copper cooling coils and iron connections---} electrolytic reaction). The EM service people told us that if we ever used acid to clean the lines that they would no longer warranty the microscope. The PVC lines are perfectly clean. (J. Bozzola)
Another material you may wish to consider is called PEX, which is a cross-linked polyethylene. I don't know too much about its characteristics, except that it's very smooth inside, which should retard crud accumulation and it's more opaque than white pvc. It may be worth checking out. (P. Grover)
As an FYI, if you have copper based and iron based materials mixed in the same system, the iron will corrode preferentially through galvanic coupling, not the copper. In fact, the iron becomes a sacrificial anode protecting the copper from corrosion. Any time you have those two materials in the same system, you should have dielectric couplings between the two or you will actively corrode the ferrous based material. Regarding acid cleaning, you should know what your deposits are in your piping before deciding how to clean them. As an FYI, copper will generally corrode at pH's below about 6.3 or so. There are low pH cleaners that can be used with copper, but they contain corrosion inhibitors. (D. Jones)
If you intend to clean the lines with acid, I suggest PVC, since Cu can be etched at low pH. In addition to floating some dichlorophene for algal control, we add a corrosion inhibitor. We have been using a Mo-based formula, which was available from Aqua Labs on the East coast and from Skasol on the West coast, so find a distributor in your area. I think that either Aqua or Skasol would be able to give you that info.(B. Tivol)
Along these lines, I noticed a lot more copper leaching into the lines when I used deionized water than regular or filtered tap water. Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze intended for cars should have corrosion inhibitors and should be suitable for this purpose. I have still gotten green particles regardless of whether antifreeze was used or not and have regularly made it a practice to clean the filters in the lines once a semester. (Jerry Calvin)
I have used the ethylene/glycol/water mix through clear PVC piping between my circulator and TEM and vacuum coating unit for six years now. There is a long run of the piping in daylight. Absolutely no algae Problem. Use 50/50 ethylene glycol*/water and PVC pipes and you will never have a problem again. Make sure that the chiller/recirculator manufacturer OKs the use of ethylene glycol but I wouldn't anticipate any problem. Even 20/80 EG/Water will work well. (Ted Dunn)
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============================== 17, 21 -- From dsherman-at-purdue.edu Sat Feb 18 10:40:08 2006 17, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 17, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1IGe8dl014778 17, 21 -- for {microscopy-at-microscopy.com} ; Sat, 18 Feb 2006 10:40:08 -0600 17, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 17, 21 -- Sat, 18 Feb 2006 11:40:08 -0500 17, 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 ; 17, 21 -- Sat, 18 Feb 2006 16:40:08 +0000 17, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 17, 21 -- Date: Sat, 18 Feb 2006 11:40:07 -0500 17, 21 -- Subject: Cooling lines-responses 17, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 17, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 17, 21 -- Message-ID: {C01CB817.D64C%dsherman-at-purdue.edu} 17, 21 -- Thread-Topic: Cooling lines-responses 17, 21 -- Thread-Index: AcY0qfmyOEGiJKCdEdqwBAARJN08Mg== 17, 21 -- Mime-version: 1.0 17, 21 -- Content-type: text/plain; 17, 21 -- charset="US-ASCII" 17, 21 -- Content-transfer-encoding: 7bit 17, 21 -- X-OriginalArrivalTime: 18 Feb 2006 16:40:08.0197 (UTC) FILETIME=[FA697350:01C634A9] ==============================End of - Headers==============================
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Email: soleimanij-at-tbzmed.ac.ir Name: Jafar Soleimani Rad
Organization: University
Title-Subject: [Filtered] scale bar
Question: Hi we are using A LEO 906 TEM, scale bar is not registered in the microfilms. We are having problem to calculating the real sizes. any help in this matter is appreciated.
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 rpowell-at-nanoprobes.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Filtered] Re: [Microscopy]: NTA-nanogold for His tagged protein in EM?
Question: [Commercial disclaimer - I work for Nanoprobes, and we make NTA-Ni(II)-NanogoldĆ]
Hello Milton:
Here are a few recent references for EM using NTA-Ni(II)-Nanogold, with links to articles on our online newsletter that describe them:
(1) Wolfe, C. L.; Warrington, J. A.; Treadwell, L., and Norcum, M. T.: A three-dimensional working model of the multienzyme complex of aminoacyl-tRNA synthetases based on electron microscopic placements of tRNA and proteins. J. Biol. Chem., 280, 38870-38878 (2005).
(2) Bumba, L.; Tichy, M.; Dobakova, M.; Komenda, J., and Vacha, F.: Localization of the PsbH subunit in photosystem II from the Synechocystis 6803 using the His-tagged NińNTA Nanogold labeling. J. Struct. Biol., 152, 28-35 (2005).
There are two more recent references in which the reagent was used to label polyhistidine-tagged components of viral capsids:
(3) Chatterji, A.; Ochoa, W. F.; Ueno, T.; Lin T., and Johnson, J. E.: A virus-based nanoblock with tunable electrostatic properties. Nano Lett., 5, 597-602 (2005).
(4) Collins, R. F.; Frye, S. A.; Balasingham, S.; Ford, R. C.; Tonjum, T., and Derrick, J. P.: Interaction with type IV pili induces structural changes in the bacterial outer membrane secretin PilQ. J. Biol. Chem., 280, 18923-18930 (2005).
Details about the reagent are available on our web site:
Catalog and general info: http://www.nanoprobes.com/NTAgold.html
Product info and instructions: http://www.nanoprobes.com/Inf2080.html
We have found that the complexes of NTA-Ni(II)-Nanogold bound to polyhis-tagged proteins hold together well during chromtographic separations (gel filtration), implying that they should be injectable.
Hope some of these are helpful,
Rick Powell Nanoprobes, Inc. www.nanoprobes.com
At 05:09 PM 2/17/2006, you wrote: Question: Has anynone had success using NTA-nanogold to disclose the location of His-tagged proteins by electron microscopy? I would like to attach the nanogold this way before injecting the protein into a cell. An alternative would be to apply gold labelled primary anti-His after fixation and permeabilization.
This Question was submitted to Ask-A-Microscopist by (grmitch-at-netzero.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, February 15, 2006 at 20:08:01 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both grmitch-at-netzero.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: grmitch-at-netzero.com Name: Linda Mitchell
Organization: BPS Environmental Center
Education: K-8 Grade Grammar School
Location: Birmingham, Michigan USA
Title: Microorganisms!
Question: I will be teaching a unit on microscopic life to 5th graders next month, March (quite a cold month here in Michigan). My question for you: What is the best way to keep my microorganisms alive throughout the program? We obtain our samples directly from the 10 acres of the property here at the nature center. One of the samples is from our pond area and the second from the swamp woods area. We have no difficulty in finding a healthy sample, yet the problem that we often encounter is keeping them alive. We have supplies - lab pans, microscopes, even an aquarium that is our "indoor pond" when the water ices over. Do you have any feeding, climate tips that would help in these areas? This is a program that is enjoyed by both the staff and students - they are so amazed by what they find. Thanks for your input.
Millennium Chemicals (a Lyondell Company), the world's second largest producer of Titanium Dioxide, is seeking a microscopist with experience in the areas of Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM) and X-ray Diffraction (XRD) to support R&D, Commercial and Manufacturing activities at its Baltimore Research Center.
EDUCATIONAL REQUIREMENTS
Minimum M.S. in chemistry, mineralogy, material science or similar field, with 7-10 years experience in an industrial R&D environment preferred.
DESCRIPTION:
The primary responsibility of the position is the application of advanced microscopic techniques to investigate properties, mineralogy, phase distribution, morphology and structure/function relationships of pigmentary and catalytic TiO2 particles, catalysts and polymers. In addition to conducting research and and support activities, the candidate will be responsible for oversight and maintenance of a state-of-the-art microscopy laboratory that includes:
*Olympus SZX7 stereoscope *Olympus BX51 Optical Light Microscope *Amray 1930 SEM with EDS (scheduled for replacement in 2006) *Jeol 2000 FX2 with EDS and STEM *Veeco Nanoscope 3A Dimension 3100 AFM *Panalytical X'pert Pro XRD *RMC PT-X Ultramicrotome with CR-X Cryosectioning system *Gatan Model 691 Precision Ion Polishing System *SPE Plasma Prep II Plasma Etcher/Asher *Denton sputterer/coater
SALARY
Salary is commensurate with education and experience. Millennium Chemicals (A Lyondell Company) offers a competitive benefits package including relocation.
For more information and to submit a resume online, please visit our website:
Millennium Chemicals (a Lyondell Company) is an Equal Opportunity Employer M/F/D/V
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==============================Original Headers============================== 20, 24 -- From Laurine.Ottmar-at-millenniumchem.com Mon Feb 20 06:02:30 2006 20, 24 -- Received: from edcpap01.lyo.com (mail3.lyondell.com [161.16.0.77]) 20, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1KC2Ucr017418 20, 24 -- for {microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 06:02:30 -0600 20, 24 -- Received: from edcexp02.lyondell.com ([161.16.33.40]) 20, 24 -- by edcpap01.lyo.com (8.13.4/8.13.4) with ESMTP id k1KC2Nb9003328 20, 24 -- for {microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 06:02:23 -0600 20, 24 -- Received: from amnhhtv01.millenniumchem.com ([172.19.0.46]) by edcexp02.lyondell.com with Microsoft SMTPSVC(5.0.2195.6713); 20, 24 -- Mon, 20 Feb 2006 06:02:24 -0600 20, 24 -- Subject: Employment Opportunity 20, 24 -- To: microscopy-at-microscopy.com 20, 24 -- X-Mailer: Lotus Notes Release 5.0.11 July 24, 2002 20, 24 -- Message-ID: {OF1550363E.97FA6230-ON8525711A.006B77CF-8525711B.0042227A-at-millenniumchem.com} 20, 24 -- From: Laurine.Ottmar-at-millenniumchem.com 20, 24 -- Date: Mon, 20 Feb 2006 07:02:22 -0500 20, 24 -- X-MIMETrack: Serialize by Router on AMNHHTV01/HUB/SRV/MIC(Release 5.0.11 |July 24, 2002) at 20, 24 -- 02/20/2006 07:02:24 AM 20, 24 -- MIME-Version: 1.0 20, 24 -- Content-type: text/plain; charset=iso-8859-1 20, 24 -- X-OriginalArrivalTime: 20 Feb 2006 12:02:24.0991 (UTC) FILETIME=[83317EF0:01C63615] 20, 24 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=0 adultscore=0 adjust=0 reason=mlx engine=3.1.0-06021401 definitions=3.0.0-06022000 20, 24 -- X-Proofpoint-OriginalSender: Laurine.Ottmar-at-millenniumchem.com 20, 24 -- Content-Transfer-Encoding: 8bit 20, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1KC2Ucr017418 ==============================End of - Headers==============================
one thing that occurs to me is that you will probably have vinyl flooring in your darkroom. This may be a major source of static and it may be possible to get some advice on low static flooring. I know we had some installed in one of our microscope cubicles and it didn't seem to be tremendously expensive at the time.
If you think that the floor is part of the problem then it might be worth checking whether the soles of your shoes are man made (create static) or leather.
If you want to go for a tried and tested electronics industry route look for ionisers (rather than de-ionisers) I did a simple Google search on ioniser and static and came up with a few using keywords "ionisers static" such as http://www.buystatic.com/static_eliminators.htm?ioniser.htm
I was hoping that you could get away with a domestic ioniser - the sort they sell to simulate sea or mountain air and supposedly gets rid of headaches. But apparently they aren't much good at eliminating static whereas the industrial ones are.
Good luck
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: tonygr-at-MIT.EDU
I have a friend who seems particularly prone to this even when the humidity is not very low - she can produce amazing patterns of sparks and spirals on a negative simply by waving her hands over them!
The simple way of getting rid of electrostatic problems in a semiconductor company like mine is to 'borrow' an earth strap and mat from the test area. It solved the problem completely. I guess you may have to buy them..
} ------------------------------------------------------------------- } --------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } AmericaTo Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserverOn-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html------------ } ---------------------------------------------------------------- } } This is a perennial problem, I know, but does anyone have any } effective "pet" solutions to the problem of charging and } subsequent electrostatic arcing of TEM film in the extremely dry } air we get in New England typically in the winter, and some other } people get at other seasons of the year? } } The discharges can occur at almost any stage of the handling, from } getting the film out of the vendor's packing, loading and } unloading in the microscope carriers and loading in the developing } racks (we use Lucite ones). We have considered a humidifier in } the darkroom, but that would involve blocking off the ventilation } (we have make-up air positively blown into to room and exhaust } actively sucked out) to prevent our moisture from being simply } blown away. } } Any hints would be welcome, but switching to digital imaging is } one that is not going to happen. } } Tony. } } ************************************* } Anthony J. Garratt-Reed M.A. D.Phil. } MIT Room #13-1027 } 77 Massachusetts Avenue } Cambridge, Massachusetts 02139-4307 } USA } } Tel: (617) 253-4622 } Fax: (617) 258-6478 } ************************************* } } } ==============================Original } Headers==============================7, 26 -- From tonygr-at-MIT.EDU } Sat Feb 18 07:54:11 2006 } 7, 26 -- Received: from biscayne-one-station.mit.edu (BISCAYNE-ONE- } STATION.MIT.EDU [18.7.7.80]) } 7, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k1IDsB7m0033107, 26 -- for {microscopy-at-microscopy.com} ; Sat, 18 } Feb 2006 07:54:11 -0600 } 7, 26 -- Received: from outgoing.mit.edu (OUTGOING-AUTH.MIT.EDU } [18.7.22.103])7, 26 -- by biscayne-one-station.mit.edu } (8.12.4/8.9.2) with ESMTP id k1IDp3gq004012 } 7, 26 -- for {microscopy-at-microscopy.com} ; Sat, 18 Feb 2006 } 08:51:03 -0500 (EST) } 7, 26 -- Received: from [192.168.1.47] (pool-72-70-109- } 28.bstnma.east.verizon.net [72.70.109.28]) } 7, 26 -- (authenticated bits=0) } 7, 26 -- (User authenticated as tonygr-at-ATHENA.MIT.EDU) } 7, 26 -- by outgoing.mit.edu (8.13.1/8.12.4) with ESMTP id } k1IDosBT0052947, 26 -- (version=TLSv1/SSLv3 cipher=DHE-RSA- AES256- } SHA bits=256 verify=NOT) } 7, 26 -- for {microscopy-at-microscopy.com} ; Sat, 18 Feb 2006 } 08:51:01 -0500 (EST) } 7, 26 -- Message-ID: {43F7263D.9010808-at-mit.edu} } 7, 26 -- Date: Sat, 18 Feb 2006 08:50:53 -0500 } 7, 26 -- From: Tony Garratt-Reed {tonygr-at-MIT.EDU} } 7, 26 -- User-Agent: Thunderbird 1.5 (Windows/20051201) } 7, 26 -- MIME-Version: 1.0 } 7, 26 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} } 7, 26 -- Subject: TEM film electrostatic discharging } 7, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 7, 26 -- Content-Transfer-Encoding: 7bit } 7, 26 -- X-Spam-Score: 1.217 } 7, 26 -- X-Spam-Level: * (1.217) } 7, 26 -- X-Spam-Flag: NO } 7, 26 -- X-Scanned-By: MIMEDefang 2.42 } ==============================End of - } Headers==============================
==============================Original Headers============================== 10, 35 -- From malcolm.haswell-at-sunderland.ac.uk Mon Feb 20 09:22:15 2006 10, 35 -- Received: from max1.sunderland.ac.uk (max1.sunderland.ac.uk [157.228.29.83]) 10, 35 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1KFMEwG029654 10, 35 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 09:22:15 -0600 10, 35 -- Received: (qmail 10493 invoked from network); 20 Feb 2006 15:22:10 -0000 10, 35 -- Received: from localhost (127.0.0.1) 10, 35 -- by max1.sunderland.ac.uk with SMTP; 20 Feb 2006 15:22:10 -0000 10, 35 -- Received: from max1.sunderland.ac.uk ([127.0.0.1]) 10, 35 -- by localhost (max1.sunderland.ac.uk [127.0.0.1]) (amavisd-new, port 10024) 10, 35 -- with SMTP id 09230-09 for {Microscopy-at-microscopy.com} ; 10, 35 -- Mon, 20 Feb 2006 15:22:06 +0000 (GMT) 10, 35 -- Received: (qmail 10462 invoked by uid 516); 20 Feb 2006 15:22:06 -0000 10, 35 -- Received: from [157.228.37.117] (HELO hermes.sunderland.ac.uk) (157.228.37.117) 10, 35 -- by max1.sunderland.ac.uk (qpsmtpd/0.28) with ESMTP; Mon, 20 Feb 2006 15:22:06 +0000 10, 35 -- Received: from sunderland.ac.uk (localhost [127.0.0.1]) 10, 35 -- by hermes.sunderland.ac.uk 10, 35 -- (iPlanet Messaging Server 5.2 Patch 2 (built Jul 14 2004)) 10, 35 -- with ESMTP id {0IUZ00MBGS0VEP-at-hermes.sunderland.ac.uk} for 10, 35 -- Microscopy-at-microscopy.com; Mon, 20 Feb 2006 15:22:07 +0000 (GMT) 10, 35 -- Received: from [157.228.15.253] by hermes.sunderland.ac.uk (mshttpd); Mon, 10, 35 -- 20 Feb 2006 15:22:07 +0000 (GMT) 10, 35 -- Date: Mon, 20 Feb 2006 15:22:07 +0000 (GMT) 10, 35 -- From: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} 10, 35 -- Subject: Re: [Microscopy] TEM film electrostatic discharging 10, 35 -- To: tonygr-at-MIT.EDU, Microscopy MSA {Microscopy-at-microscopy.com} 10, 35 -- Message-id: {b19b04b19e96.b19e96b19b04-at-sunderland.ac.uk} 10, 35 -- MIME-version: 1.0 10, 35 -- X-Mailer: iPlanet Messenger Express 5.2 Patch 2 (built Jul 14 2004) 10, 35 -- Content-type: text/plain; charset=us-ascii 10, 35 -- Content-language: en 10, 35 -- Content-transfer-encoding: 7BIT 10, 35 -- Content-disposition: inline 10, 35 -- X-Accept-Language: en 10, 35 -- Priority: normal 10, 35 -- X-Virus-Scanned: by iCritical at max1.sunderland.ac.uk ==============================End of - Headers==============================
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Mon Feb 20 10:38:12 2006 8, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1KGcBCj008019 8, 31 -- for {microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 10:38:11 -0600 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-4.tower-78.messagelabs.com!1140453489!7816629!1 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 28862 invoked from network); 20 Feb 2006 16:38:09 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-4.tower-78.messagelabs.com with SMTP; 20 Feb 2006 16:38:09 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 8, 31 -- Mon, 20 Feb 2006 16:38:08 +0000 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 31 -- Content-class: urn:content-classes:message 8, 31 -- MIME-Version: 1.0 8, 31 -- Content-Type: text/plain; 8, 31 -- charset="us-ascii" 8, 31 -- Subject: Re: TEM film electrostatic discharging 8, 31 -- Date: Mon, 20 Feb 2006 16:38:08 -0000 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E028190-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: TEM film electrostatic discharging 8, 31 -- Thread-Index: AcY2MdESLSSjQkN8S4i066gcnb8TQQACVscg 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 20 Feb 2006 16:38:08.0664 (UTC) FILETIME=[07FDA980:01C6363C] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1KGcBCj008019 ==============================End of - Headers==============================
Advice needed on TEM sample prep for cultured cells.
We are growing skin fibroblasts in culture and would like to examine the cells with transmission electron microscopy. To date, our results have been disappointing. We grew the cells on Thermanox coverslips, fixed for 1 hr at room temp in cacodylate-buffered 1.5% glutaraldehyde, 1.5% paraformaldehyde. After buffer washes, the cells were post-fixed with 1% osmium tetroxide, then washed, dehydrated in acetone and embedded in Epon/Araldite.
The cells look OK in general, but the membranes (both plasma membrane and internal membranes) are all rather fuzzy and indistinct. The quality of the ultrastructure is much poorer that we obtain with tissues prepared using almost identical proceures. One would think that since the cells are only a monolayer, preservation would be excellent.
If you have experience with TEM of cultured cells and have obtained good results, I would appreciate any advice you can give me to get better quality ultrastructure.
Please respond to me directly at katzm-at-health.missouri.edu
Thanks,
Martin L. Katz, Ph.D. Professor Ophthalmology, Pathobiology, Neurosciences, Genetics Mason Eye Institute University of Missouri School of Medicine Columbia, Missouri 65212 Phone (573) 882-8480 FAX (573) 884-4100 katzm-at-health.missouri.edu http://www.muhealth.org/~ophthalmology/Katz.htm
==============================Original Headers============================== 9, 23 -- From TindallR-at-missouri.edu Mon Feb 20 13:19:57 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 (8.12.11/8.12.8) with ESMTP id k1KJJv9B019566 9, 23 -- for {microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 13:19:57 -0600 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, 20 Feb 2006 13:19:57 -0600 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="iso-8859-1" 9, 23 -- Subject: TEM: Cell culture preparation 9, 23 -- Date: Mon, 20 Feb 2006 13:19:56 -0600 9, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849D7E-at-UM-EMAIL09.um.umsystem.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: TEM: Cell culture preparation 9, 23 -- Thread-Index: AcY2UqHzjBChwr/YROWHPWjAaQBpkg== 9, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 9, 23 -- To: {microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 20 Feb 2006 19:19:57.0367 (UTC) FILETIME=[A2D42C70:01C63652] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1KJJv9B019566 ==============================End of - Headers==============================
You should be able to dispense with the formalin (not "paraformaldehyde" -- that's the solid from which fresh formalin is made). Monolayers of cells generally do not need formalin. Add 1% tannic acid to both the glut and osmium. Mallinckrodt 1764; the monomeric form seems to work better. You may only need the tannic acid in one of the fixes, probably the glut, but putting it in both the fixes won't hurt. You may need to fix for 2 hours, but one hour should be enough. Note: this is for high-resolution SEM, where any holes in the plasma membrane are blatant, and it works well up to mags } 100kX (and higher). Also, you could try growing the cells on glass coverslips, and digesting off the coverslip with HF after fixation, but that's probably not relevant to your problem. Good luck.
Phil
} Advice needed on TEM sample prep for cultured cells. } } We are growing skin fibroblasts in culture and } would like to examine the cells with } transmission electron microscopy. To date, our } results have been disappointing. We grew the } cells on Thermanox coverslips, fixed for 1 hr at } room temp in cacodylate-buffered 1.5% } glutaraldehyde, 1.5% paraformaldehyde. After } buffer washes, the cells were post-fixed with 1% } osmium tetroxide, then washed, dehydrated in } acetone and embedded in Epon/Araldite. } } The cells look OK in general, but the membranes } (both plasma membrane and internal membranes) } are all rather fuzzy and indistinct. The } quality of the ultrastructure is much poorer } that we obtain with tissues prepared using } almost identical proceures. One would think } that since the cells are only a monolayer, } preservation would be excellent. } } If you have experience with TEM of cultured } cells and have obtained good results, I would } appreciate any advice you can give me to get } better quality ultrastructure. } } Please respond to me directly at katzm-at-health.missouri.edu } } Thanks, } } Martin L. Katz, Ph.D. } Professor } Ophthalmology, Pathobiology, Neurosciences, Genetics } Mason Eye Institute } University of Missouri School of Medicine } Columbia, MissouriÝ 65212 } Phone (573) 882-8480 } FAX (573) 884-4100 } katzm-at-health.missouri.edu } Ýhttp://www.muhealth.org/~ophthalmology/Katz.htm -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 5, 25 -- From oshel1pe-at-cmich.edu Mon Feb 20 13:57:20 2006 5, 25 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 5, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1KJvK7J029432 5, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 13:57:20 -0600 5, 25 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 5, 25 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k1KKaO4l026727; 5, 25 -- Mon, 20 Feb 2006 15:36:24 -0500 5, 25 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 5, 25 -- Mon, 20 Feb 2006 14:57:16 -0500 5, 25 -- Mime-Version: 1.0 5, 25 -- Message-Id: {f06230903c01fcdfc338c-at-[141.209.160.132]} 5, 25 -- In-Reply-To: {200602201925.k1KJPv8q027648-at-ns.microscopy.com} 5, 25 -- References: {200602201925.k1KJPv8q027648-at-ns.microscopy.com} 5, 25 -- Date: Mon, 20 Feb 2006 14:57:16 -0500 5, 25 -- To: TindallR-at-missouri.edu 5, 25 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 5, 25 -- Subject: Re: [Microscopy] TEM: Cell culture preparation 5, 25 -- Cc: Microscopy-at-microscopy.com 5, 25 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 5, 25 -- X-OriginalArrivalTime: 20 Feb 2006 19:57:17.0011 (UTC) FILETIME=[D9C2BA30:01C63657] 5, 25 -- X-CanItPRO-Stream: default 5, 25 -- X-Spam-Score: -4 () L_EXCH_MF 5, 25 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 5, 25 -- Content-Transfer-Encoding: 8bit 5, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1KJvK7J029432 ==============================End of - Headers==============================
Many (and I do mean many) years ago I fixed cultures of retinal pigment epithelial cells for TEM. I used a routine cacodylate-buffered glut. fix, no formaldehyde although that should not matter. I did put 1% tannic acid in the fix, both glut and osmium and it helped show the ECM but I don' think it should be essential. I grew the cells in ordinary culture dishes, no thermanox c'slips and embedded in Epon substitute. My guess is that your membranes look fuzzy due to less than optimal fixation of lipids (old glut and/or osmium?) or extraction of lipids (too long in dehydration?). Why are you using acetone and not ethanol? It is my understanding that acetone removes lipids faster than EtOH but I have also heard that the converse it true. Certainly too long in dehydration can degrade ultrastructure. Also, add 2mM Ca ions to the fix to help stabilize the lipids.
Geoff
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 7, 32 -- From mcauliff-at-umdnj.edu Mon Feb 20 14:25:41 2006 7, 32 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 7, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1KKPfHB006954 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 14:25:41 -0600 7, 32 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 7, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 058694BDF9 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 15:25:41 -0500 (EST) 7, 32 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 7, 32 -- by zix03.umdnj.edu (Proprietary) with ESMTP id 60A724BE2D 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 15:25:40 -0500 (EST) 7, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 7, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 7, 32 -- id {0IV0009015TECC-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 7, 32 -- for microscopy-at-msa.microscopy.com; Mon, 20 Feb 2006 15:25:40 -0500 (EST) 7, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 7, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 7, 32 -- 2004)) with ESMTP id {0IV0001KL62R6J-at-Polaris.umdnj.edu} ; Mon, 7, 32 -- 20 Feb 2006 15:25:39 -0500 (EST) 7, 32 -- Date: Mon, 20 Feb 2006 15:24:54 -0500 7, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 7, 32 -- Subject: Re: [Microscopy] TEM: Cell culture preparation 7, 32 -- In-reply-to: {200602201921.k1KJLIov021601-at-ns.microscopy.com} 7, 32 -- To: TindallR-at-missouri.edu, 7, 32 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 7, 32 -- Message-id: {43FA2596.8050405-at-umdnj.edu} 7, 32 -- MIME-version: 1.0 7, 32 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 7, 32 -- Content-transfer-encoding: 7BIT 7, 32 -- X-Accept-Language: en-us, en 7, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 7, 32 -- Gecko/20040804 Netscape/7.2 (ax) 7, 32 -- References: {200602201921.k1KJLIov021601-at-ns.microscopy.com} ==============================End of - Headers==============================
I see that several people are using spam filters on this list. I have just received e-mail from 2 people supposedly on this list asking me to click on a link to verify that I am really sending them a non-spam e-mail. No way am I going to click on a link from someone I don't know! What ARE these people thinking?
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, 29 -- From mcauliff-at-umdnj.edu Mon Feb 20 15:10:53 2006 6, 29 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 6, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1KLArhW017219 6, 29 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 15:10:53 -0600 6, 29 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 6, 29 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 1F1151C327F 6, 29 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 16:10:53 -0500 (EST) 6, 29 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 6, 29 -- by zix01.umdnj.edu (Proprietary) with ESMTP id BEE40A7B41 6, 29 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 16:10:51 -0500 (EST) 6, 29 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 29 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 29 -- id {0IV000H017WH9Y-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 29 -- for microscopy-at-msa.microscopy.com; Mon, 20 Feb 2006 16:10:51 -0500 (EST) 6, 29 -- Received: from [127.0.0.1] ([10.138.2.240]) 6, 29 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 29 -- 2004)) with ESMTP id {0IV0001G880C6J-at-Polaris.umdnj.edu} for 6, 29 -- microscopy-at-msa.microscopy.com; Mon, 20 Feb 2006 16:07:24 -0500 (EST) 6, 29 -- Date: Mon, 20 Feb 2006 16:06:39 -0500 6, 29 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 29 -- Subject: spam filters or ?? 6, 29 -- To: MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 29 -- Message-id: {43FA2F5F.7050807-at-umdnj.edu} 6, 29 -- MIME-version: 1.0 6, 29 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 29 -- Content-transfer-encoding: 7BIT 6, 29 -- X-Accept-Language: en-us, en 6, 29 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 29 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
We recently saw a TIRF system demonstrated, and like other laser based TIRF systems, it seems to use a small peripheral arc aperture to allow the laser light through to part of the outside periphery of the 1.45NA objective. Why would you not use a complete ring annulus instead of an arc to illuminate the entire outside periphery of the objective? If you simply put the correct sized annulus into the field stop in the fluorescence path, would you get TIRF? Also, what is the theoretical advantage of using a laser as opposed to an arc illuminator. Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology U-3125 91 N. Eagleville Rd. University of Connecticut Storrs, CT 06269 860-486-2200 860-486-4331 (fax)
==============================Original Headers============================== 3, 19 -- From david.knecht-at-uconn.edu Mon Feb 20 15:32:18 2006 3, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 3, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1KLWITb026790 3, 19 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 15:32:18 -0600 3, 19 -- Received: from [137.99.47.163] (d47h163.public.uconn.edu [137.99.47.163]) 3, 19 -- by mail2.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k1KLW0m26333 3, 19 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 20 Feb 2006 16:32:00 -0500 3, 19 -- Mime-Version: 1.0 (Apple Message framework v746.2) 3, 19 -- Content-Transfer-Encoding: 7bit 3, 19 -- Message-Id: {8E45F94A-D08D-4926-9B2F-ECE8BF9D3920-at-uconn.edu} 3, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 3, 19 -- To: Microscopy-at-msa.microscopy.com 3, 19 -- From: David Knecht {david.knecht-at-uconn.edu} 3, 19 -- Subject: TIRF setup 3, 19 -- Date: Mon, 20 Feb 2006 16:32:02 -0500 3, 19 -- X-Mailer: Apple Mail (2.746.2) 3, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 3, 19 -- X-UConn-MailScanner: Found to be clean 3, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
For the floor, there is a spray that can be put on it to temporarily stop electrostatic buildup. I'm sorry, but I can't remember the name of it, but I have used it in the past.
For electronic use, I found that taking my shoes off solves static discharge problems. There is enough moisture on my feet to stop any discharges.
-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: malcolm.haswell-at-sunderland.ac.uk [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Monday, February 20, 2006 7:28 AM To: Walck-at-SouthBayTech.com
Tony
one thing that occurs to me is that you will probably have vinyl flooring in your darkroom. This may be a major source of static and it may be possible to get some advice on low static flooring. I know we had some installed in one of our microscope cubicles and it didn't seem to be tremendously expensive at the time.
If you think that the floor is part of the problem then it might be worth checking whether the soles of your shoes are man made (create static) or leather.
If you want to go for a tried and tested electronics industry route look for ionisers (rather than de-ionisers) I did a simple Google search on ioniser and static and came up with a few using keywords "ionisers static" such as http://www.buystatic.com/static_eliminators.htm?ioniser.htm
I was hoping that you could get away with a domestic ioniser - the sort they sell to simulate sea or mountain air and supposedly gets rid of headaches. But apparently they aren't much good at eliminating static whereas the industrial ones are.
Good luck
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: tonygr-at-MIT.EDU
Some of the solutions suggested seem to be based on the idea that the operator is picking up the static charge. I think the problem is much more likely to be caused by static on the film being discharged while unloading the exposed negatives. I now unload film wearing rubber gloves and rarely get a static discharge pattern on my film.
Ralph Common Dept. of Physiology Michigan State University
==============================Original Headers============================== 3, 25 -- From rcommon-at-msu.edu Mon Feb 20 16:01:59 2006 3, 25 -- Received: from sys15.mail.msu.edu (sys15.mail.msu.edu [35.9.75.115]) 3, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1KM1xUa013444 3, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Feb 2006 16:01:59 -0600 3, 25 -- Received: from [35.9.122.125] (helo=emlab) 3, 25 -- by sys15.mail.msu.edu with esmtpsa (Exim 4.52 #1) 3, 25 -- (TLSv1:RC4-MD5:128) 3, 25 -- id 1FBJ6U-0001zU-V8 3, 25 -- for Microscopy-at-microscopy.com; Mon, 20 Feb 2006 17:01:59 -0500 3, 25 -- From: "Ralph Common" {rcommon-at-msu.edu} 3, 25 -- To: {Microscopy-at-microscopy.com} 3, 25 -- Subject: TEM film electrostatic discharging 3, 25 -- Date: Mon, 20 Feb 2006 17:02:37 -0500 3, 25 -- Message-ID: {001401c63669$5dc89410$7d7a0923-at-msu.edu} 3, 25 -- MIME-Version: 1.0 3, 25 -- Content-Type: text/plain; 3, 25 -- charset="iso-8859-1" 3, 25 -- Content-Transfer-Encoding: 7bit 3, 25 -- X-Priority: 3 (Normal) 3, 25 -- X-MSMail-Priority: Normal 3, 25 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) 3, 25 -- In-Reply-To: {200602201528.k1KFSxtR005488-at-ns.microscopy.com} 3, 25 -- Importance: Normal 3, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 3, 25 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
Interesting question.... As I understand it the colors seen in thin sections are caused by destructive interference. The equation: Wavelength = 2nt/m describes the color seen. n = refractive index of film t is thickness (nm) M is any integer.
If so it seems that as refractive index decreases, wavelength will also decrease for the same thickness film.
I hope this help and thank you for giving me a chance to thumb through my old reference books, it brought back memories....
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==============================Original Headers============================== 9, 18 -- From frank.karl-at-degussa.com Tue Feb 21 07:09:25 2006 9, 18 -- Received: from mailout1.degussa.com (mailout1.degussa.com [193.100.56.173]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LD9OXV002077 9, 18 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 07:09:25 -0600 9, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 9, 18 -- by mailout1.degussa.com (8.13.3/8.13.3/Debian-6) with SMTP id k1LD9B0P024169; 9, 18 -- Tue, 21 Feb 2006 14:09:17 +0100 9, 18 -- In-Reply-To: {200602172210.k1HMA6nj031675-at-ns.microscopy.com} 9, 18 -- Subject: Re: [Microscopy] viaWWW: microtome section thickness 9, 18 -- To: leis-at-biochem.mpg.de, microscopy-at-msa.microscopy.com 9, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 9, 18 -- Message-ID: {OFC1F9625B.D0797714-ON8525711C.004779CF-8525711C.004839F5-at-degussa.com} 9, 18 -- From: frank.karl-at-degussa.com 9, 18 -- Date: Tue, 21 Feb 2006 08:08:54 -0500 9, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 9, 18 -- 02/21/2006 07:09:18 AM 9, 18 -- MIME-Version: 1.0 9, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I ignore these suspicious requests as well. I guess those folk don't get much mail now.
Dave
-----Original Message----- X-from: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu] Sent: 20 February 2006 21:15 To: David Patton
Dear List;
I see that several people are using spam filters on this list. I have just received e-mail from 2 people supposedly on this list asking me to click on a link to verify that I am really sending them a non-spam
e-mail. No way am I going to click on a link from someone I don't know! What ARE these people thinking?
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 **********************************************
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==============================Original Headers============================== 20, 33 -- From David.Patton-at-uwe.ac.uk Tue Feb 21 08:55:34 2006 20, 33 -- Received: from mailapp01.uwe.ac.uk (mailapp01.uwe.ac.uk [164.11.132.61]) 20, 33 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1LEtVGe013689 20, 33 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 08:55:33 -0600 20, 33 -- Received: from (164.11.132.60) by mailapp01.uwe.ac.uk via smtp 20, 33 -- id 4a43_94e119d4_a2ea_11da_89e4_0002b3c946e4; 20, 33 -- Tue, 21 Feb 2006 14:58:56 +0000 20, 33 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 20, 33 -- (egen-uwe01.campus.ads.uwe.ac.uk [164.11.249.121]) 20, 33 -- by mta01.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 20, 33 -- 2005)) with ESMTP id {0IV1006AELGIO1-at-mta01.uwe.ac.uk} for 20, 33 -- microscopy-at-msa.microscopy.com; Tue, 21 Feb 2006 14:55:30 +0000 (GMT) 20, 33 -- Date: Tue, 21 Feb 2006 14:55:29 +0000 20, 33 -- From: David Patton {David.Patton-at-uwe.ac.uk} 20, 33 -- Subject: RE: [Microscopy] spam filters or ?? 20, 33 -- To: microscopy-at-msa.microscopy.com 20, 33 -- Message-id: {F247F674896BE243AD8263C5280E2BDB022BB804-at-egen-uwe01} 20, 33 -- MIME-version: 1.0 20, 33 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.6944.0 20, 33 -- Content-type: text/plain; charset=us-ascii 20, 33 -- Content-class: urn:content-classes:message 20, 33 -- Thread-topic: [Microscopy] spam filters or ?? 20, 33 -- Thread-index: AcY2YzRIJ0lBMgRET9a4rhY9wkWYYgAk2+uQ 20, 33 -- X-MS-Has-Attach: 20, 33 -- X-MS-TNEF-Correlator: 20, 33 -- X-NAI-Spam-Level: * 20, 33 -- X-NAI-Spam-Score: 1.5 20, 33 -- X-NAI-Spam-Rules: 2 Rules triggered 20, 33 -- NAI_ZIP_CODE=4, BAYES_00=-2.5 20, 33 -- X-NAIMIME-Disclaimer: 1 20, 33 -- X-NAIMIME-Modified: 1 20, 33 -- Content-Transfer-Encoding: 8bit 20, 33 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1LEtVGe013689 ==============================End of - Headers==============================
I have heard other people report situation like this and I, too, have experienced the same thing. Often time (not always) monolayer cells showed very little membrane contrast, even though the tissue processed same way had no problem. The problem with monolayer cells seemed random regardless what types of cells were being processed. More interestingly, I have not seen this problem with cell suspensions. In the past, I tried to use freshly made OsO4 once when I had low membrane contrast problem with monolayer cells, and that helped. But I still do not understand why the problem only occurs in monolayer cells. I do not think it is a reagent penetration issue, nor a problem of inadequate processing protocol. Is it possible that some kind of coating material used in culture makes it harder for OsO4 to react with lipid molecules? Thank you.
Hong Emory EM
==============================Original Headers============================== 5, 22 -- From hyi-at-emory.edu Tue Feb 21 10:06:29 2006 5, 22 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LG6S2K024016 5, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Feb 2006 10:06:28 -0600 5, 22 -- Received: from [170.140.232.202] (localhost [127.0.0.1]) 5, 22 -- by virginia.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k1LG6OIf028249 5, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Feb 2006 11:06:28 -0500 (EST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v622) 5, 22 -- Message-Id: {5a5232b7e407dc1cfb2b84946528c088-at-emory.edu} 5, 22 -- Content-Type: text/plain; 5, 22 -- charset=ISO-8859-1; 5, 22 -- format=flowed 5, 22 -- To: microscopy-at-microscopy.com 5, 22 -- From: Hong Yi {hyi-at-emory.edu} 5, 22 -- Subject: Re: [Microscopy] TEM: Cell culture preparation 5, 22 -- Date: Tue, 21 Feb 2006 11:06:21 -0500 5, 22 -- X-Mailer: Apple Mail (2.622) 5, 22 -- X-imss-version: 2.037 5, 22 -- X-imss-result: Passed 5, 22 -- X-imss-approveListMatch: *-at-emory.edu 5, 22 -- Content-Transfer-Encoding: 8bit 5, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1LG6S2K024016 ==============================End of - Headers==============================
I process monolayers of cells frequently for TEM. The problem of low membrane contrast is due to the lipids being extracted away during dehydration and embedding. Even membranes "stabilized" with OsO4 can be extracted with the long processing times used during "standard fixations". I typically dehydrate in EtOH for 1-2 minutes per EtOH grade, with my samples dehydrated from 100% water to 100% EtOH in 15-20 minutes. I also keep the time in liquid resin to a minimum...my whole embedding protocol takes less than 3 hours. After I shortened my times considerably, my membranes started looking very nice and crisp! Good Luck -Eugene
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology II Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
} Hi, Randy: } } I have heard other people report situation like this and I, } too, have experienced the same thing. Often time (not always) monolayer } cells showed very little membrane contrast, even though the tissue } processed same way had no problem. The problem with monolayer cells } seemed random regardless what types of cells were being processed. More } interestingly, I have not seen this problem with cell suspensions. In } the past, I tried to use freshly made OsO4 once when I had low membrane } contrast problem with monolayer cells, and that helped. But I still do } not understand why the problem only occurs in monolayer cells. I do not } think it is a reagent penetration issue, nor a problem of inadequate } processing protocol. Is it possible that some kind of coating material } used in culture makes it harder for OsO4 to react with lipid } molecules? } Thank you. } } Hong } Emory EM } } } } } ==============================Original Headers============================== } 5, 22 -- From hyi-at-emory.edu Tue Feb 21 10:06:29 2006 } 5, 22 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) } 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LG6S2K024016 } 5, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Feb 2006 10:06:28 -0600 } 5, 22 -- Received: from [170.140.232.202] (localhost [127.0.0.1]) } 5, 22 -- by virginia.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k1LG6OIf028249 } 5, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Feb 2006 11:06:28 -0500 (EST) } 5, 22 -- Mime-Version: 1.0 (Apple Message framework v622) } 5, 22 -- Message-Id: {5a5232b7e407dc1cfb2b84946528c088-at-emory.edu} } 5, 22 -- Content-Type: text/plain; } 5, 22 -- charset=ISO-8859-1; } 5, 22 -- format=flowed } 5, 22 -- To: microscopy-at-microscopy.com } 5, 22 -- From: Hong Yi {hyi-at-emory.edu} } 5, 22 -- Subject: Re: [Microscopy] TEM: Cell culture preparation } 5, 22 -- Date: Tue, 21 Feb 2006 11:06:21 -0500 } 5, 22 -- X-Mailer: Apple Mail (2.622) } 5, 22 -- X-imss-version: 2.037 } 5, 22 -- X-imss-result: Passed } 5, 22 -- X-imss-approveListMatch: *-at-emory.edu } 5, 22 -- Content-Transfer-Encoding: 8bit } 5, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1LG6S2K024016 } ==============================End of - Headers============================== } }
==============================Original Headers============================== 6, 20 -- From krueger.eugene-at-mayo.edu Tue Feb 21 10:56:47 2006 6, 20 -- Received: from mail1.mayo.edu (mail1.mayo.edu [129.176.212.12]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LGuloa001993 6, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Feb 2006 10:56:47 -0600 6, 20 -- Received: from mhro1a.mayo.edu ([129.176.212.53]) 6, 20 -- by mail1.mayo.edu with ESMTP; 21 Feb 2006 10:56:46 -0600 6, 20 -- X-BrightmailFiltered: true 6, 20 -- X-Brightmail-Tracker: AAAAAA== 6, 20 -- Received: from excsrv01.mayo.edu (excsrv01.mayo.edu [129.176.235.101]) by mhro1a.mayo.edu with ESMTP for Microscopy-at-microscopy.com; Tue, 21 Feb 2006 10:56:46 -0600 6, 20 -- Received: by excsrv01.mayo.edu with Internet Mail Service (5.5.2657.72) 6, 20 -- id {FMHXTMZG} ; Tue, 21 Feb 2006 10:53:21 -0600 6, 20 -- Message-Id: {FC1B895EA707A940A6BD5F74106FAA9A269497-at-excsrv80.mayo.edu} 6, 20 -- From: "Krueger, Eugene W." {krueger.eugene-at-mayo.edu} 6, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 6, 20 -- Subject: Re: TEM: Cell culture preparation 6, 20 -- Date: Tue, 21 Feb 2006 10:56:42 -0600 6, 20 -- MIME-Version: 1.0 6, 20 -- X-Mailer: Internet Mail Service (5.5.2657.72) 6, 20 -- Content-Type: text/plain; 6, 20 -- charset="iso-8859-1" ==============================End of - Headers==============================
On Feb 18, 2006, at 5:54 AM, tonygr-at-MIT.EDU wrote:
} This is a perennial problem, I know, but does anyone have any } effective "pet" solutions to the problem of charging and subsequent } electrostatic arcing of TEM film in the extremely dry air we get in } New England typically in the winter, and some other people get at } other seasons of the year? } } The discharges can occur at almost any stage of the handling, from } getting the film out of the vendor's packing, loading and unloading in } the microscope carriers and loading in the developing racks (we use } Lucite ones). We have considered a humidifier in the darkroom, but } that would involve blocking off the ventilation (we have make-up air } positively blown into to room and exhaust actively sucked out) to } prevent our moisture from being simply blown away. } } Any hints would be welcome, but switching to digital imaging is one } that is not going to happen. } Dear Tony, When removing the film from the envelopes, keep it in a pack, so none of the films rubs along any other, then lift the cardboard without rubbing. If you can maintain contact with a grounded metal surface--the water pipes are good--this will help a lot. Try not to let your lab coat (or other clothing for that matter) to move along your body or other items of clothing. Always separate films by bending the top one away from the others, then lifting. Trade your lucite racks for metal ones. These procedures reduced, but did not completely eliminate, the problem when I was in Albany. When using LoDose film in total darkness, I could at least see the discharges when they occurred. 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 Feb 21 13:32:32 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LJWVHu013187 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 13:32:32 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 5719E35BB9 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 11:32:31 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 2D62435A54 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 11:32:30 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602181354.k1IDsSQp003552-at-ns.microscopy.com} 4, 22 -- References: {200602181354.k1IDsSQp003552-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {8eab6d7bd81e0fcbc6fc662ef0c1c9fa-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] TEM film electrostatic discharging 4, 22 -- Date: Tue, 21 Feb 2006 11:40:34 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
We have a vintage Sorvall MT-1 ultramicrotome available for the cost of shipping. Manual included. Has no stereomicroscope with it. Great for semithin plastic sections. All manual operation. It is headed for the junk pile if no one adopts it.
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 3, 22 -- From gwe-at-ufl.edu Tue Feb 21 14:18:40 2006 3, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 3, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LKIeL1023349 3, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 14:18:40 -0600 3, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 3, 22 -- (authenticated bits=0) 3, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k1LKIacU4337810 3, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 15:18:37 -0500 3, 22 -- Message-ID: {43FB759D.6050002-at-ufl.edu} 3, 22 -- Date: Tue, 21 Feb 2006 15:18:37 -0500 3, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 3, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 3, 22 -- X-Accept-Language: en-us, en 3, 22 -- MIME-Version: 1.0 3, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 3, 22 -- Subject: MT-1 for free 3, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 22 -- Content-Transfer-Encoding: 7bit 3, 22 -- X-Spam-Status: hits=-0.904, required=5, tests=BAYES_30 3, 22 -- X-UFL-Spam-Status: hits=-0.904, required=5, tests=BAYES_30 3, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
I was asked to post my embedding schedule, so here it is:
When embedding monolayers (~60 - 90% confluent) in epoxy resin (I prefer Quetol 651) I follow this schedule:
graded series of EtOH, 25%, 50%, 70%, 95%, 100%, 100%, total dehydration time 15-20' 100%EtOH:Quetol 651 (1:1), ~25' 100% Quetol 651, ~50' 100% Quetol 651, ~100' fresh 100% Quetol 651, then into 60C oven
I do my processing in a 6 or 12 well plate that sits on a shelf in the hood (NOT on a rotator), and I change to a new 6 or 12 well plate between changes of 100% EtOH. I also use minimal volumes of resin to decrease extraction of lipid components. If you use a more viscous resin, times may need to be adjusted longer. Good Luck!
-Eugene Eugene W. A. Krueger Sr. Research Technologist in Cell Biology II Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
==============================Original Headers============================== 5, 20 -- From krueger.eugene-at-mayo.edu Tue Feb 21 14:47:11 2006 5, 20 -- Received: from mail2.mayo.edu (mail2.mayo.edu [129.176.212.15]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1LKlBv7003576 5, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Feb 2006 14:47:11 -0600 5, 20 -- Received: from mhro1a.mayo.edu ([129.176.212.53]) 5, 20 -- by mail2.mayo.edu with ESMTP; 21 Feb 2006 14:47:10 -0600 5, 20 -- X-BrightmailFiltered: true 5, 20 -- X-Brightmail-Tracker: AAAAAA== 5, 20 -- Received: from excsrv01.mayo.edu (excsrv01.mayo.edu [129.176.235.101]) by mhro1a.mayo.edu with ESMTP for Microscopy-at-microscopy.com; Tue, 21 Feb 2006 14:47:10 -0600 5, 20 -- Received: by excsrv01.mayo.edu with Internet Mail Service (5.5.2657.72) 5, 20 -- id {FMHXW34R} ; Tue, 21 Feb 2006 14:43:45 -0600 5, 20 -- Message-Id: {FC1B895EA707A940A6BD5F74106FAA9A269498-at-excsrv80.mayo.edu} 5, 20 -- From: "Krueger, Eugene W." {krueger.eugene-at-mayo.edu} 5, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 5, 20 -- Subject: embedding schedule for monolayers 5, 20 -- Date: Tue, 21 Feb 2006 14:47:07 -0600 5, 20 -- MIME-Version: 1.0 5, 20 -- X-Mailer: Internet Mail Service (5.5.2657.72) 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="iso-8859-1" ==============================End of - Headers==============================
On Feb 18, 2006, at 2:32 PM, soleimanij-at-tbzmed.ac.ir wrote:
} we are using A LEO 906 TEM, scale bar is not registered in the } microfilms. We are having problem to calculating the real sizes. any } help in this matter is appreciated. } Dear Jafar, I would calibrate the magnification(s) of interest either with a cross-grating replica or, preferably, a Mag*I*Cal, both of which are available from many supply houses. If you need sizes on prints, there is a calibration slide that can be photographed in an enlarger, with which you can determine the ratio of print sizes to negative sizes; I don't know where to get this, but it must be widely available. 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
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Dear List;
I see that several people are using spam filters on this list. I have just received e-mail from 2 people supposedly on this list asking me to click on a link to verify that I am really sending them a non-spam e-mail. No way am I going to click on a link from someone I don't know! What ARE these people thinking?
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 **********************************************
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==============================Original Headers============================== 23, 27 -- From milesd-at-us.ibm.com Tue Feb 21 20:05:07 2006 23, 27 -- Received: from e3.ny.us.ibm.com (e3.ny.us.ibm.com [32.97.182.143]) 23, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1M257Q0001451 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 20:05:07 -0600 23, 27 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 23, 27 -- by e3.ny.us.ibm.com (8.12.11/8.12.11) with ESMTP id k1M255Vc016162 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:05 -0500 23, 27 -- Received: from d01av01.pok.ibm.com (d01av01.pok.ibm.com [9.56.224.215]) 23, 27 -- by d01relay04.pok.ibm.com (8.12.10/NCO/VERS6.8) with ESMTP id k1M2562i220526 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:06 -0500 23, 27 -- Received: from d01av01.pok.ibm.com (loopback [127.0.0.1]) 23, 27 -- by d01av01.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k1M255LU008116 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:05 -0500 23, 27 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 23, 27 -- by d01av01.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k1M2540P008094 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:04 -0500 23, 27 -- In-Reply-To: {200602202111.k1KLBUjW018639-at-ns.microscopy.com} 23, 27 -- Subject: Re: [Microscopy] spam filters or ?? 23, 27 -- To: Microscopy-at-MSA.Microscopy.Com 23, 27 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 23, 27 -- Message-ID: {OF22CCB88F.52878652-ON8525711D.000B000E-8525711D.000B7274-at-us.ibm.com} 23, 27 -- From: Darrell Miles {milesd-at-us.ibm.com} 23, 27 -- Date: Tue, 21 Feb 2006 21:05:03 -0500 23, 27 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 6.5.4FP2 HF4|December 20, 2005) at 23, 27 -- 02/21/2006 21:05:04 23, 27 -- MIME-Version: 1.0 23, 27 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I'm looking for a reliable scanner for TEM Micrographs. For the past 4 years I used an old HP scanner that was very good with 35mm and TEM micrographs, unfortunate this scanner is no longer working, now I need a scanner that can capture a reliable image for routine everyday micrographs; in order to shorten darkroom time. Any subjection?
Thanks in advance
Omayra Velez Life Cell Branchburg, NJ
==============================Original Headers============================== 5, 18 -- From mayas003-at-yahoo.com Tue Feb 21 22:58:21 2006 5, 18 -- Received: from web84109.mail.dcn.yahoo.com (web84109.mail.dcn.yahoo.com [209.73.179.120]) 5, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1M4wK2N012775 5, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 22:58:20 -0600 5, 18 -- Received: (qmail 63646 invoked by uid 60001); 22 Feb 2006 04:58:20 -0000 5, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 18 -- s=s1024; d=yahoo.com; 5, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 5, 18 -- b=l3I3qltvjW7C7MFLKskZiUVjJbPQ1cydee1f6xuI3x0YEOtyb1XLN+xZX7pO4ne8HOWyjClZfM4jfomwRvPZ1aIimFvPffyyv5mgzkS65fZVHYy/qLK30Kq3Ft6gNcfeVuPoHIlCg5oBNKSj3juJH9BgGJ/Hx0brg5OPd0fZ8gY= ; 5, 18 -- Message-ID: {20060222045820.63644.qmail-at-web84109.mail.dcn.yahoo.com} 5, 18 -- Received: from [68.239.240.252] by web84109.mail.dcn.yahoo.com via HTTP; Tue, 21 Feb 2006 20:58:20 PST 5, 18 -- Date: Tue, 21 Feb 2006 20:58:20 -0800 (PST) 5, 18 -- From: Omayra Velez {mayas003-at-yahoo.com} 5, 18 -- Subject: Scanner for TEM Micrographs 5, 18 -- To: Microscopy-at-MSA.Microscopy.Com 5, 18 -- MIME-Version: 1.0 5, 18 -- Content-Type: text/plain; charset=iso-8859-1 5, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Like microscopes, it really is the more you pay the better the result (even my kids Ł60 DX5 microscope has 250x magnification and a CMOS 'camera').
However, from my experience even going to Ł2,000 for a scanner, the result of a negative or slide is sometimes not as good as the original if you really start magnifying it up (although I haven't tried the latest 4000 to 5000 dpi top of the range Nikon slide scanners). This is mainly due to film grain size optical effects that even Digital GEM doesn't seem to wholly eliminate (it is just a software image processing system even if embedded into the scanner hardware & optimised for the scanner). Grain size optical effects are such that a 400ASA negative colour scan at 2,700dpi can produce an appallingly unusable image that image processing can't really save - garbage in garbage out (whereas a reflective scan of the 6x4" print produces a very reasonable A4 image). Higher resolutions approaching or exceeding the grain size of the file (e.g. 4000 dpi and above) are supposed to reduce this problem considerably but Nikon type slide scanners are expensive and very limited in negative size options. In practice many problems in image quality are as much due to ease of magnifying a digital image compared to the negative - a few clicks and you have a 'print' the size of a wall.
However the scanned slide image from lower ASA 50-100 slides at 4000dpi is probably better on the VDU than most standard (not enlargements) prints made from the negative/slide and it can easily go up to A3 in output to a printer, which is probably all that really matters (and you still have the negative in the likely event the next generation scanners will get even better for the price). However enlargement prints from the negative may be better than a magnified scanned image. Naturally if you want to zoom in on a negative, it would have been better to do this on the TEM in the first place and take another picture instead (wise after the event).
In practice it might also be that our eyes prefer a fuzzy analogue gradation of colour rather than the very defined little squares of a pixellated image at the same resolution. We are also good at discerning contrast. Also remember that digital camera's often do some image processing during capture (e.g. noise reduction, colour correction and sometimes even sharpen) so you have to work on the image after scanning to get the same result. Top of the range scanner software and hardware can do this 'automatically' using calibration targets and so forth (e.g. Digital GEM, SHO, ICE and Silverfast Ai). This scan processing can triple scan times to 10 minutes or more, but can save far more than that on manual Photoshop type processing times afterwards (and make a better job of it). Note that we can only discern 191 grey levels so 8-bit (256 grey levels) B&W scans be fine for most uses except perhaps image analysis - and this really reduced image file size.
So the scanned images can look very good at A3 and on a 22" monitor, and if your negative archive is going the way of my collection of 1950's 35mm home slides and literally disintegrating into brown mush, anything is an improvement. Surprisingly the twain software can also have an effect on scanned quality in some cases (cheaper slide scanners often benefit from using Silverfast SE over the cheaper bundled software).
To scan many large negatives at high resolution via a 5000dpi Creo type hi-resolution flatbed scanner would need an investment of Ł12k to Ł45 and the massive files would be hard to manage and archive without image size reduction and compression which renders the whole procedure rather pointless. Given the cost it would seem to be preferable to pay someone to scan the odd few negatives you need scanned at this resolution with their Creo. Hi-resolution drum scanners up to 1500dpi cost nearer Ł70k but if your collection is something like NASA's archive that price can be well worth paying (see their results at http://grin.hq.nasa.gov). A 2k x 2k or greater pixel image digital camera system for a TEM costs around Ł20k to Ł70k.
You can get pretty good results from the new breed of sub-Ł1,000 flatbed scanners though, particularly with large format negatives (i.e. TEM). Have a look at the reviews of these semi-pro flatbed scanners on the net (around Ł300 to 800 to buy). Below is an excellent link for the Canon 9950F that could be used for 4x5" or 9 x 6cm film. It also compares the results with the similar Epson 4990 Photo.
http://www.photo-i.co.uk
I use the Canon 9950F (Ł260) at home for 35mm negatives and slides. The Canon's main weakness is that it's twain interface can only scan to the frame sizes (i.e. not to my square Kodak 125 slides, where it chops on the top and bottom of the slides to fit it to 35mm - a real pain). With Silverfast SE [twain] also provided, you can scan any film size. However Canon won't share the FARE dust removal technology with Silverfast so FARE isn't supported - not a problem with B&W negatives as FARE and ICE only work with colour - although you can scan B&W negatives in 'colour' with FARE/ICE and convert to B&W in Photoshop - but 'results may vary'. I have upgraded to Silverfast Ai for $115 for my Canon 9950F which adds in auto multi-slide scans (but not FARE). Canon no longer manufacture dedicated slide scanners as they feel this flatbed is as good.
At work I use the Epson 4990 (Ł300). It has a better twain interface and has an A4 negative scan feature that can scan any smaller negatives in multiples if they don't fit the standard frames provided. Silverfast SE also provided supports ICE infra-red dust removal - but dust removal can reduce image quality a little if the negative isn't that dusty. I use a standard rubber bulb to blow dust off before scanning - air jets canisters are gone in a day and can squirt propellant over the film. It's image quality is identical to Canon 9950F for 35mm slides, although the Epson can only scan 8 slides at one go to the Canon's 12. Scan time is similar for both (very slow with FARE/ICE - about 10 minutes a 35mm slide). However both these flatbeds are really 2,400 dpi not 4800 dpi as claimed - if you scan 35mm at 4800 dpi the image quality is virtually identical the 2,400 dpi scan. However you can't regularly scan full TEM at 4,800 dpi and above as the image size would be near 1 Gb - we scan at 800 to 1,200 dpi mainly - although you can of course scan small areas at 2,400 dpi resolution for 'enlargements'. All uses say they are happy with the TEM scan quality.
If you keep the negative anyway, I would have thought being able to print to A3 size is adequate for most purposes. The cheap Canon 9950F flatbed has replaced my 2,700 dpi SCSI Scanwit 2740s dedicated slide scanner at home (largely as it's so much faster and easier to use). I have to say the image quality of these flatbeds is a little out of focus (or 'soft' as we call it) at full magnification, but the careful use of USM (unsharp mask) in Photoshop can improve this a lot. But they are fine up to A3 at least (from a 35mm slide). Flatbed scans always need a little more Twain and post scan tweeking than dedicated film scanners. Leave things like USM and colour balance to Photoshop but use the twain interface to set things like brightness in dark negatives and dust ICE/FARE removal.
These scanners can come with expensive Silverfast Ai and targets though (sometimes as a Pro version) or you can upgrade at silverfast.com. But a lot of this is for accurate colour (not TEM's strong point) - although Silverfast is a powerful & complicated Twain interface.
When going from 2,700 dpi of the old slide/negative scanners up to the 4,000 dpi of modern Nikon type slide/negative scanners many users report far better image quality, and put it down to reduced effects from grain aliasing e.g. http://hardware.mcse.ms/message144915.html and presumably the same will be true of TEM negatives and the latest 4,000 dpi semi-pro flatbed scanners (that can take 4"x5") as well. At these resolutions film grain is also very apparent though, as 35mm film grain has a lower dpi.
There's also dedicated & cheap large film scanners like the Epson F3200 going to 4x5" film but again the resolution of the F3200 is quoted at 'only' 3200dpi, plus it got a duff review : http://www.photo-i.co.uk/Reviews/interactive/Scanners/Epson_F3200/page-1.htm
In fact it is probably a little better for image quality (prior to USM) than the flatbeds for large B&W negatives, but you may be limited to the frame sizes so check if you have 6x9cm negatives rather than proper Ľ plate. It seems that it can scan any size smaller than the largest frame - but certainly there's no A4 negative or reflective scanning.
Have a look at: http://www.photoscientia.co.uk/Grain.htm for discussions of grain size.
Have a look at http://www.datamind.co.uk/merchant/resolution.htm for some chat on pixel and image file sizes.
Hope this is of some use.
Regards
Keith
PS. This a bit large to post on the listerver, but it gave me something to do while travelling for 4 hours on the train to & from work each day. This follows on from similar threads about scanner a few months ago.
----- Original Message ----- X-from: {mayas003-at-yahoo.com} To: {keith.morris-at-ucl.ac.uk} Sent: Wednesday, February 22, 2006 5:02 AM
I put my EM negatives on a light box, mask off all other areas with black paper (so 'extra' light does not mislead the light meter), copy with my digital camera, then invert the image to a positive and convert to grayscale in PhotoShop. Fast, easy and excellent results.
Geoff
mayas003-at-yahoo.com wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 7, 31 -- From mcauliff-at-umdnj.edu Wed Feb 22 09:02:53 2006 7, 31 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 7, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1MF2qZ5007257 7, 31 -- for {microscopy-at-msa.microscopy.com} ; Wed, 22 Feb 2006 09:02:52 -0600 7, 31 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 7, 31 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 586FC4BE3C 7, 31 -- for {microscopy-at-msa.microscopy.com} ; Wed, 22 Feb 2006 09:02:52 -0600 (CST) 7, 31 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 7, 31 -- by zix02.umdnj.edu (Proprietary) with ESMTP id 2EB6F4BE34 7, 31 -- for {microscopy-at-msa.microscopy.com} ; Wed, 22 Feb 2006 09:02:51 -0600 (CST) 7, 31 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 7, 31 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 7, 31 -- id {0IV300E01GF9ER-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 7, 31 -- for microscopy-at-msa.microscopy.com; Wed, 22 Feb 2006 10:02:50 -0500 (EST) 7, 31 -- Received: from [127.0.0.1] ([10.138.2.240]) 7, 31 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 7, 31 -- 2004)) with ESMTP id {0IV300H8HGBMQ5-at-Polaris.umdnj.edu} ; Wed, 7, 31 -- 22 Feb 2006 09:59:46 -0500 (EST) 7, 31 -- Date: Wed, 22 Feb 2006 09:59:01 -0500 7, 31 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 7, 31 -- Subject: Re: [Microscopy] Scanner for TEM Micrographs 7, 31 -- In-reply-to: {200602220459.k1M4xdY3014788-at-ns.microscopy.com} 7, 31 -- To: mayas003-at-yahoo.com, MicroscopyListserver {microscopy-at-msa.microscopy.com} 7, 31 -- Message-id: {43FC7C35.5050601-at-umdnj.edu} 7, 31 -- MIME-version: 1.0 7, 31 -- Content-type: text/plain; format=flowed; charset=us-ascii 7, 31 -- Content-transfer-encoding: 7BIT 7, 31 -- X-Accept-Language: en-us, en 7, 31 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 7, 31 -- Gecko/20040804 Netscape/7.2 (ax) 7, 31 -- References: {200602220459.k1M4xdY3014788-at-ns.microscopy.com} ==============================End of - Headers==============================
I am not sure why these would not be legitimate. I have gotten such requests that cited my message of some date and some subject, so they already have my e-mail address. I can easily envision an anti-spam service where a person must respond to such a challenge to get added to a white-list of senders. I have replied to a couple of those posts as instructed and have not been asked to re-register.
However, I also sent them a note that such a service is not very compatible with membership in a list server like this. Everyone who ever posts to the list will get challenged to register their e-mail address. I doubt that many will. And if I declined to register and continued to get these registration requests, I would probably add that person to my junk mailers list.
If someone halfway around the world doesn't want to accept the knowledge that gets shared via this list then that is their problem. They should not use such a challenge system on the e-mail address they use for this or other listservers.
from previous replies on the list it seems to come down to about 3-4 basic types of film scanner: 1. Very expensive dedicated drum scanners 2. Standard dedicated film scanners like the Nikon 3. A combined flatbed and glassless scanner (Agfa used to make the Duo- scan) 4. Standard flatbed scanner with glass.
I think that Keith has covered everything but the 3. combined scanner. I use a Microtek Scanmaker 8700 and find it useful because there is no glass between the optics and the film which should reduce Newton's Rings, dust and finger marks. The new Microtek is cheaper and more powerful than mine was. It handles true 3200 x 6400 dpi scans, 4.2 Dmax negatives, 48 bit colour and sells for under 700 UK pounds or about 600 US dollars. I think it still comes with Silversoft scanning software and Digital ICE (scratch and dust reducing software/hardware). This should give up to 16x enlargements with reasonably dense negatives up to sizes of 10x 8 inches or more (about 4 of our 4489 negatives at once). See http://www.microtekusa.com/smi900.html
A lot depends on the size and nature of your negatives as to whether they will fit into a particular scanner. If you produce a lot of dense negatives (usually not a problem with biological samples) then maybe you will need more than 4.2 Dmax.
I don't know of any of the off-the-shelf scanners which come with a ready made holder for the larger e.m. negatives so you may have to get something made up even if it's just out of bits of cardboard or plastic.
Good luck
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: keith.morris-at-ucl.ac.uk
While your observation is valid, in some cases those subscribing to listservers have no control over how spam is filtered. Our email goes through the corporate servers located over a mile away and through filters set up by the corporate IT group. Some of their filters are rather exasperating for us in the microscopy group. For example, ALL embedded images from incoming emails are deleted by the corporate filters. What we get in its place is the marker [IMAGE]. If we need to have images sent to us, they must come as attachments. Again, this is out of our control. So too with spam. Fortunately our corporate filters direct anything it thinks is spam into a "spam folder". We can go through the emails in the folder at our leisure and if it is from someone we know we can mark it and add it to the list of allowed senders. But unlike the systems you are describing, we, as the recipient, must accept the email. Nothing is sent to the poster as far as I can tell. This is one of the reasons I like having the [Microscopy] tag on the subject line. It makes it easier to pick out the real emails in my horrendous list of spam in the morning.
My point in all this is we should not be too hard on those posters since they may not have any control over the situation and may not even know it is happening.
Sharon Goresh Chemist Engelhard Corp Research Center Iselin, NJ
wesaia-at-iastate.ed u To 02/22/06 10:13 AM sharon.goresh-at-engelhard.com cc
Please respond to Subject wesaia-at-iastate.ed [Microscopy] Re: spam filters or ?? u
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
I am not sure why these would not be legitimate. I have gotten such requests that cited my message of some date and some subject, so they already have my e-mail address. I can easily envision an anti-spam service where a person must respond to such a challenge to get added to a white-list of senders. I have replied to a couple of those posts as instructed and have not been asked to re-register.
However, I also sent them a note that such a service is not very compatible with membership in a list server like this. Everyone who ever posts to the list will get challenged to register their e-mail address. I doubt that many will. And if I declined to register and continued to get these registration requests, I would probably add that person to my junk mailers list.
If someone halfway around the world doesn't want to accept the knowledge that gets shared via this list then that is their problem. They should not use such a challenge system on the e-mail address they use for this or other listservers.
Thanks for extra interesting info. We actually have an elderly Agfa Duo-Scan T2550 here (your option 3) that cost Ł1,000's in its day - it has a SCSI interface and is attached to an old Apple laptop with no CD-RW and 10mb network connection only (for 'historical reasons' not my choice, although the Apple brigade like it). It's so noisy and such a pain to use that we bought the Epson 4990 Photo just for the convenience of fast USB2 scans to Windows XP. Our T2500 DuoScan is 'only' rated at 2,500 x 2,500 dpi (but being a Pro scanner it really means it - consumer hardware often has more 'optimistic' 4,800 dpi resolution claims). I'll have to compare TEM negative scans on both the DuoScan and Epson 4990 at full res, but here XP Pro PC users have voted with their feet and mostly use the Epson 4990 as they only need 1,200 dpi max anyway and its sitting in their office.
As you no doubt know Agfa, Heidelberg, and Fuji have completely withdrawn from the pro flatbed scanner market 'due to market conditions'. It now appears that Minolta Konica (e.g. Dimage/Dynax) from the consumer market have joined them. Microtek flatbeds used to be very solidly engineered, my old SCSI flatbed (c2000) had a metal sub-chassis and always hurt my back when I lifted it out from under the desk - they don't seem to be quite so highly regarded these days, although many reviewers don't seem to spend as much time tweaking the scanned images via the twain interface and Photoshop as they should to get the best from the scanner.
Keith
For more details of top end flatbed pro scanners have a look at: http://www.imaging-resource.com/NEWS/1098478219.html If have to ask the price of a Creo you can't afford one http://www.flatbed-scanner-review.org/ This site is now 'pay per view' for recent reviews
---------------------------------------------------------- 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: {malcolm.haswell-at-sunderland.ac.uk} To: {keith.morris-at-ucl.ac.uk} Sent: Wednesday, February 22, 2006 3:18 PM
RE membrane contrast, We thought it was due to a trace oxidant in the ethanol that re-oxidizes the osmium deposited in the cells. It would only show up in cultured cells, because one has such a huge bath of ethanol and such a small mass of biologicals.
I have a solution for the problem, I will send you (Martin). It should be on our web site, but I shall have to look to see. It keeps the membranes - the basic nature of the artifact is still unknown. carol
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-- __ 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-bgnet.bgsu.edu http://www.bgsu.edu/departments/biology/facilities/MnM ___________________________________________________________________________
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Recently a colleague and I were doing some SEM on... uh... wallaby droppings (no, seriously!) and we came across some objects that neither of us have seen before. I've posted some images and an EDS spectrum of these mysterious things at:
http://www.mta.ca/dmf/download/ehrman/mystery.htm
As you can see from the spectrum, they appear to be organic, and in fact beam current for EDS does damage them pretty easily. They appear to be hollow, and they survived (or possibly are an artifact of?) wallaby digestion, desiccation, 5% KOH treatment, sonication, and the usual SEM type prep. Any ideas? Has anybody seen these sorts of things NOT coming out the rear end of a wallaby?
As usual, thanks in advance for your interest,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
We've been having a devil of a time fixing intact e18.5 day mouse limbs for TEM. We would like to keep them intact, including all regions between the end of the digits and elbow, so the total length is about 2 mm. We are specifically interested in tendon and tendon sheath. We would like to keep the epithelium intact, but so far our best fixation results have been following the removal of the digit tips and making small slices through the skin which allows better penetration of fixative and infiltration of epoxy. I would remove the skin, but the tendons are so close to the surface that I'd prefer not to disturb them. We've tried extending the fixation time in 1.5% glut/1.5% paraformaldehyde to days, extending the OsO4 to days; we've used a laboratory Pelco microwave to assist and we are about out of ideas. Fixation with OsO4 is not complete (white areas in the middle of the limb) and infiltration with epoxy is usually not complete (though the microwave does seem to help here). Perhaps there is a trick to this, or maybe some exotic fixative I should try? Suggestions are welcome!
Many thanks in advance,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
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Chack out a selection of excellent scanner reviews at Imaging Resources.
http://www.imaging-resource.com/SCAN1.HTM
John Mardinly
-----Original Message----- X-from: keith.morris-at-ucl.ac.uk [mailto:keith.morris-at-ucl.ac.uk] Sent: Wednesday, February 22, 2006 4:52 AM To: Mardinly, John
Dear Omayra,
Like microscopes, it really is the more you pay the better the result (even my kids Ł60 DX5 microscope has 250x magnification and a CMOS 'camera').
However, from my experience even going to Ł2,000 for a scanner, the result of a negative or slide is sometimes not as good as the original if you really start magnifying it up (although I haven't tried the latest 4000 to 5000 dpi top of the range Nikon slide scanners). This is mainly due to film grain size optical effects that even Digital GEM doesn't seem to wholly eliminate (it is just a software image processing system even if embedded into the scanner hardware & optimised for the scanner). Grain size optical effects are such that a 400ASA negative colour scan at 2,700dpi can produce an appallingly unusable image that image processing can't really save - garbage in garbage out (whereas a reflective scan of the 6x4" print produces a very reasonable A4 image). Higher resolutions approaching or exceeding the grain size of the file (e.g. 4000 dpi and above) are supposed to reduce this problem considerably but Nikon type slide scanners are expensive and very limited in negative size options. In practice many problems in image quality are as much due to ease of magnifying a digital image compared to the negative - a few clicks and you have a 'print' the size of a wall.
However the scanned slide image from lower ASA 50-100 slides at 4000dpi is probably better on the VDU than most standard (not enlargements) prints made from the negative/slide and it can easily go up to A3 in output to a printer, which is probably all that really matters (and you still have the negative in the likely event the next generation scanners will get even better for the price). However enlargement prints from the negative may be better than a magnified scanned image. Naturally if you want to zoom in on a negative, it would have been better to do this on the TEM in the first place and take another picture instead (wise after the event).
In practice it might also be that our eyes prefer a fuzzy analogue gradation of colour rather than the very defined little squares of a pixellated image at the same resolution. We are also good at discerning contrast. Also remember that digital camera's often do some image processing during capture (e.g. noise reduction, colour correction and sometimes even sharpen) so you have to work on the image after scanning to get the same result. Top of the range scanner software and hardware can do this 'automatically' using calibration targets and so forth (e.g. Digital GEM, SHO, ICE and Silverfast Ai). This scan processing can triple scan times to 10 minutes or more, but can save far more than that on manual Photoshop type processing times afterwards (and make a better job of it). Note that we can only discern 191 grey levels so 8-bit (256 grey levels) B&W scans be fine for most uses except perhaps image analysis - and this really reduced image file size.
So the scanned images can look very good at A3 and on a 22" monitor, and if your negative archive is going the way of my collection of 1950's 35mm home slides and literally disintegrating into brown mush, anything is an improvement. Surprisingly the twain software can also have an effect on scanned quality in some cases (cheaper slide scanners often benefit from using Silverfast SE over the cheaper bundled software).
To scan many large negatives at high resolution via a 5000dpi Creo type hi-resolution flatbed scanner would need an investment of Ł12k to Ł45 and the massive files would be hard to manage and archive without image size reduction and compression which renders the whole procedure rather pointless. Given the cost it would seem to be preferable to pay someone to scan the odd few negatives you need scanned at this resolution with their Creo. Hi-resolution drum scanners up to 1500dpi cost nearer Ł70k but if your collection is something like NASA's archive that price can be well worth paying (see their results at http://grin.hq.nasa.gov). A 2k x 2k or greater pixel image digital camera system for a TEM costs around Ł20k to Ł70k.
You can get pretty good results from the new breed of sub-Ł1,000 flatbed scanners though, particularly with large format negatives (i.e. TEM). Have a look at the reviews of these semi-pro flatbed scanners on the net (around Ł300 to 800 to buy). Below is an excellent link for the Canon 9950F that could be used for 4x5" or 9 x 6cm film. It also compares the results with the similar Epson 4990 Photo.
http://www.photo-i.co.uk
I use the Canon 9950F (Ł260) at home for 35mm negatives and slides. The Canon's main weakness is that it's twain interface can only scan to the frame sizes (i.e. not to my square Kodak 125 slides, where it chops on the top and bottom of the slides to fit it to 35mm - a real pain). With Silverfast SE [twain] also provided, you can scan any film size. However Canon won't share the FARE dust removal technology with Silverfast so FARE isn't supported - not a problem with B&W negatives as FARE and ICE only work with colour - although you can scan B&W negatives in 'colour' with FARE/ICE and convert to B&W in Photoshop - but 'results may vary'. I have upgraded to Silverfast Ai for $115 for my Canon 9950F which adds in auto multi-slide scans (but not FARE). Canon no longer manufacture dedicated slide scanners as they feel this flatbed is as good.
At work I use the Epson 4990 (Ł300). It has a better twain interface and has an A4 negative scan feature that can scan any smaller negatives in multiples if they don't fit the standard frames provided. Silverfast SE also provided supports ICE infra-red dust removal - but dust removal can reduce image quality a little if the negative isn't that dusty. I use a standard rubber bulb to blow dust off before scanning - air jets canisters are gone in a day and can squirt propellant over the film. It's image quality is identical to Canon 9950F for 35mm slides, although the Epson can only scan 8 slides at one go to the Canon's 12. Scan time is similar for both (very slow with FARE/ICE - about 10 minutes a 35mm slide). However both these flatbeds are really 2,400 dpi not 4800 dpi as claimed - if you scan 35mm at 4800 dpi the image quality is virtually identical the 2,400 dpi scan. However you can't regularly scan full TEM at 4,800 dpi and above as the image size would be near 1 Gb - we scan at 800 to 1,200 dpi mainly - although you can of course scan small areas at 2,400 dpi resolution for 'enlargements'. All uses say they are happy with the TEM scan quality.
If you keep the negative anyway, I would have thought being able to print to A3 size is adequate for most purposes. The cheap Canon 9950F flatbed has replaced my 2,700 dpi SCSI Scanwit 2740s dedicated slide scanner at home (largely as it's so much faster and easier to use). I have to say the image quality of these flatbeds is a little out of focus (or 'soft' as we call it) at full magnification, but the careful use of USM (unsharp mask) in Photoshop can improve this a lot. But they are fine up to A3 at least (from a 35mm slide). Flatbed scans always need a little more Twain and post scan tweeking than dedicated film scanners. Leave things like USM and colour balance to Photoshop but use the twain interface to set things like brightness in dark negatives and dust ICE/FARE removal.
These scanners can come with expensive Silverfast Ai and targets though (sometimes as a Pro version) or you can upgrade at silverfast.com. But a lot of this is for accurate colour (not TEM's strong point) - although Silverfast is a powerful & complicated Twain interface.
When going from 2,700 dpi of the old slide/negative scanners up to the 4,000 dpi of modern Nikon type slide/negative scanners many users report far better image quality, and put it down to reduced effects from grain aliasing e.g. http://hardware.mcse.ms/message144915.html and presumably the same will be true of TEM negatives and the latest 4,000 dpi semi-pro flatbed scanners (that can take 4"x5") as well. At these resolutions film grain is also very apparent though, as 35mm film grain has a lower dpi.
There's also dedicated & cheap large film scanners like the Epson F3200 going to 4x5" film but again the resolution of the F3200 is quoted at 'only' 3200dpi, plus it got a duff review : http://www.photo-i.co.uk/Reviews/interactive/Scanners/Epson_F3200/page-1.htm
In fact it is probably a little better for image quality (prior to USM) than the flatbeds for large B&W negatives, but you may be limited to the frame sizes so check if you have 6x9cm negatives rather than proper Ľ plate. It seems that it can scan any size smaller than the largest frame - but certainly there's no A4 negative or reflective scanning.
Have a look at: http://www.photoscientia.co.uk/Grain.htm for discussions of grain size.
Have a look at http://www.datamind.co.uk/merchant/resolution.htm for some chat on pixel and image file sizes.
Hope this is of some use.
Regards
Keith
PS. This a bit large to post on the listerver, but it gave me something to do while travelling for 4 hours on the train to & from work each day. This follows on from similar threads about scanner a few months ago.
----- Original Message ----- X-from: {mayas003-at-yahoo.com} To: {keith.morris-at-ucl.ac.uk} Sent: Wednesday, February 22, 2006 5:02 AM
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Email: sirapa-at-optonline.net Name: A. Paris
Organization: Leica Microsystems
Title-Subject: [Filtered] Glass Slide marker
Question: I am looking for an ink based glass slide marker that can be mounted to the nosepiece of an optical microscope and mark a viewed area for later observation. I found one from Ernest Fullam that marks a 3mm diamter circle but need to mark a much smaller area. Any suggestions?
This might sound like it is off-topic, but it is question that I need for the development of a possible product that would be sold in Europe and may be affected by ROHS directives and this is the first place that I thought I could get a quick answer.
Can anyone tell me whether stores are still going to be able to sell lead sinkers and lead split shot sinkers for fishing when the ROHS rules go into affect?
-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
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Tom- The protocol is on our web site, which you can find at URL: http://www.bgsu.edu/departments/biology/facilities/MnM/
If you click on Protocols under Transmission Electron Microscopy, you will find the method for in situ embedding. I speculate that the ethanol has trace oxidants in it that re-oxidize the reduced osmium in the tissue. Perhaps this only happens in tissue cultures, becuase there is so very little mass of cells compared to the bath of ethanol. It is only an idea. In any case, the use of hexyline glycol as a dehydrating agent seems to solve the problem. Carol
==============================Original Headers============================== 4, 15 -- From heckman-at-bgnet.bgsu.edu Wed Feb 22 21:11:27 2006 4, 15 -- Received: from smtp01.bgsu.edu (smtp.bgsu.edu [129.1.5.17]) 4, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1N3BRsn001768 4, 15 -- for {microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 21:11:27 -0600 4, 15 -- Received: from localhost (webmail.bgsu.edu [129.1.5.22]) 4, 15 -- by smtp01.bgsu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k1N3BQrG006644 4, 15 -- for {microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 22:11:26 -0500 (EST) 4, 15 -- MIME-Version: 1.0 4, 15 -- Date: Wed, 22 Feb 2006 22:11:26 -0500 4, 15 -- From: "heckman-at-bgnet.bgsu.edu" {heckman-at-bgnet.bgsu.edu} 4, 15 -- Message-Id: {1140664286-7372.035.108-smmsdV2.1.2-at-smtp.bgsu.edu} 4, 15 -- X-SMMS-Source: 72.240.88.3 4, 15 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 4, 15 -- Subject: Re: [Microscopy] Fwd: Re: TEM: Cell culture preparation 4, 15 -- In-Reply-To: {6.0.0.22.2.20060222200237.03db65e0-at-pop.missouri.edu} ==============================End of - Headers==============================
Dear James, being no specialist (neither in SEM-image interpretation nor SEM specimen preparation but remembering the chapter: } What izz' it { in Ultrastructural Pathology or the Chapter } What is it ? { in the Proceedings of the RMS) I only can guess:
} Nano-Soccer World Championship 2006 {: some samples of the "round leather" (as we call the } foot {-"balls" it here in Europe)...type "economy class" with holes in it for proper } aerodynamics {........no, seriousely, I do not have any invulnerable idea what those structures might be, except some kind of "vegetable" remnants including specialized perhaps symbiontic algae with a hard shell intermingled with the vegetable food wallabys perhaps are taking up........assuming that the only elements you found (cf. spectrum) are C and O (Au from sputtering?).......what about } Si { ? Perhaps you should forward your images to Phytotomists working in the field of algae.
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Recently a colleague and I were doing some SEM on... uh... wallaby droppings (no, seriously!) and we came across some objects that neither of us have seen before. I've posted some images and an EDS spectrum of these mysterious things at:
http://www.mta.ca/dmf/download/ehrman/mystery.htm
As you can see from the spectrum, they appear to be organic, and in fact beam current for EDS does damage them pretty easily. They appear to be hollow, and they survived (or possibly are an artifact of?) wallaby digestion, desiccation, 5% KOH treatment, sonication, and the usual SEM type prep. Any ideas? Has anybody seen these sorts of things NOT coming out the rear end of a wallaby?
As usual, thanks in advance for your interest,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I suppose I should have mentioned that Agfa pro 'DuoScan' scanners were rebadged Microtek scanners, and the Agfa DuoScan pretty much lives on as the Microtek ArtixScan 2500f that has 'pre-press quality' i.e. is relatively expensive at probably more than Ł2k (I can't actually find one for sale, its probably bespoke suppliers only, although it is on the Microtek.com product list).
The Microtek ArtixScan 4500tf SCSI film scanner apparently can scan up to 4 x 5" film at 2,500 dpi max, but the superb 4000 dpi Nikon film scanners (LS 5000 ED Ł900, and LS 9000ED Ł2500) are limited to 6 x 9 cm (not full 1/4 plate and above). If you use 6 x 9cm (3.5 x 2.3") film fine - 1/4 plate is 3.13 x 4.13". Many manufacturers and reviewers are not always clear on whether they are talking inches or centimetres which doesn't help. These film scanners don't have the ability to do A4 negative scans or any reflective scans at all (e.g. photographs & paper). The Nikons can apparently scan glass slides as well, if you don't have happen to have four or five optical microscopes handy.
The prosumer Epson 4990 Photo A4 flatbed [& F3200 5x4" film / 6x4" photo scanner] and the Canon 9950F's main advantage over the pro scanners is naturally they are very cheap at below Ł500, plus they can scan A4 reflective & film (or at least the Epson 4990 Photo can easily - Canon 9950F's poor twain interface limits it a bit), and therefore offer a good pixel per buck, particularly if you keep the original negatives anyway. They are also USB2 rather than pain in the ass SCSI.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: "Malcolm Haswell" {malcolm.haswell-at-sunderland.ac.uk} To: {keith.morris-at-ucl.ac.uk} ; {mayas003-at-yahoo.com} ; "Microscopy MSA" {microscopy-at-microscopy.com} Sent: Wednesday, February 22, 2006 3:14 PM
James,
I suspect that this is some silicaceous remnant of something in the wallaby's diet--something like diatom casts, plant opal phytoliths, or some related thing. Like Wolfgang, I assume (?) that the Au comes from sputtering and is simply overwhelming a silicon signal in the background. I have never personally seen a phytolith quite like these, but one of the preeminent phytolith specialists, Dr. Deborah Pearsall, is on our campus and I will forward your link to her, if you would resend it to me. I accidentally deleted the message yesterday.
I love stuff like this, especially when people ask me "What kinds of things do EM people look at?". I can now add wallaby poop to the distinguished and growing list.
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Feb 23 08:12:14 2006 10, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 10, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NECE79010968 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 08:12:14 -0600 10, 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); 10, 23 -- Thu, 23 Feb 2006 08:12:11 -0600 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="US-ASCII" 10, 23 -- Subject: RE: AW: Mystery Object 10, 23 -- Date: Thu, 23 Feb 2006 08:12:10 -0600 10, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849DB5-at-UM-EMAIL09.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: RE: AW: Mystery Object 10, 23 -- Thread-Index: AcY4gzBZ4JLS2nYZQMePbpUl3uDXVQ== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 23 Feb 2006 14:12:11.0851 (UTC) FILETIME=[23C349B0:01C63883] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1NECE79010968 ==============================End of - Headers==============================
Since it is possible that the electrostatic discharge may occur at any point in the handling of the TEM film, I recommend "foot grounders." These are straps that are put over regular footwear that keep the wearer grounded at all times. Thus, no discharge while handling film, carrying canisters, loading, etc. They can be found through any anti-ESD supplier, are easy-on, easy-off, and inexpensive enough to have a pair or two in the lab.
I have not personally tried these as ESD is not common in our lab, however, my friend in the explosives industry never goes to work without them!
-- 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============================== 4, 19 -- From raristau-at-ims.uconn.edu Thu Feb 23 08:15:47 2006 4, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NEFlOR016615 4, 19 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 08:15:47 -0600 4, 19 -- Received: from [137.99.20.175] (d20h175.public.uconn.edu [137.99.20.175]) 4, 19 -- by mail2.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k1NEFYR09634 4, 19 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 09:15:34 -0500 4, 19 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 4, 19 -- Date: Thu, 23 Feb 2006 09:15:22 -0500 4, 19 -- Subject: re: TEM film electrostatic discharging 4, 19 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 4, 19 -- To: {microscopy-at-microscopy.com} 4, 19 -- Message-ID: {C0232DAA.8A8%raristau-at-ims.uconn.edu} 4, 19 -- Mime-version: 1.0 4, 19 -- Content-type: text/plain; charset="US-ASCII" 4, 19 -- Content-transfer-encoding: 7bit 4, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 4, 19 -- X-UConn-MailScanner: Found to be clean 4, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
I received a large number of responses to this - some on the list, a larger number private. Thank you all, I have lots of food for thought.
There were several different suggestions. Many were along the lines of wearing grounding straps to ground the user, along with conductive bench tops.
The sense I have is that the main problem comes about when the fresh film is removed from the packet, and individual sheets are taken off the stack. The discharging occurs between the surface of the emulsion and the substrate of the next sheet of film. This seems to me to be unconnected with the grounding of the user, but a problem with the dryness of the film itself, and its highly non-conductive surface. The film is stored in air, and the problem is much less severe in the summer. We load the film in the film carriers before dessiccation, so the handling afterwards is minimized. I may try keeping the opened packets of film in a cabinet with a dish of water, to keep the humidity of the film up.
One responder suggested the use of MACO film (we along with, I imagine, the majority of other US users, presently use Kodak SO-163). I've seen ads for this, but never used it. Does anyone else have any experience?
Tony.
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
I am sending the following job positions for my research colleague.
Thanks.
Zhenquan Liu
Zhenquan Liu Arizona State University Center for Solid State Science S. Research Specialist PSA 213, Tempe, AZ 85-285-1704 Tel (480) 965 4512 Zhenquan.liu-at-asu.edu
Two Contract Researchers Are Needed in the Catalyst Characterization Group of Monsanto Company
Job Location: Creve Coeur Campus, St. Louis, Missouri
The following two openings (one year contract researchers) are immediately available:
1) Research associate for TEM sample preparation. Required skills: extensive experience and expertise in ultramicrotoming thin sections of catalyst powders or other nanophase materials for transmission electron microscopy observation. This job requires strong hands-on skills and new method development for unique samples. Experience with sample preparation equipments such as high vacuum carbon coating, embedding, fixing and staining biological tissues, and maintenance of sample preparation facility is highly desirable. Experience in operating SEM instruments is a plus.
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The duration of the above two jobs is one year. Since Monsanto does not directly hire contract researchers the selected candidates will work for a contract agency. Interested parties please send your resume and application letter to: Jingyue.liu-at-monsanto.com.
==============================Original Headers============================== 12, 26 -- From Zhenquan.Liu-at-asu.edu Thu Feb 23 09:29:44 2006 12, 26 -- Received: from post5.inre.asu.edu (post5.inre.asu.edu [129.219.110.120]) 12, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NFThIY010225 12, 26 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 09:29:44 -0600 12, 26 -- Received: from conversion.post5.inre.asu.edu by asu.edu (PMDF V6.1-1X6 #30769) 12, 26 -- id {0IV500701CCTAQ-at-asu.edu} for Microscopy-at-microscopy.com; Thu, 12, 26 -- 23 Feb 2006 08:29:17 -0700 (MST) 12, 26 -- Received: from EX03.asurite.ad.asu.edu 12, 26 -- (excl1-b0.asurite.ad.asu.edu [129.219.12.197]) 12, 26 -- by asu.edu (PMDF V6.1-1X6 #30769) with ESMTP id {0IV500730CCT5T-at-asu.edu} for 12, 26 -- Microscopy-at-microscopy.com; Thu, 23 Feb 2006 08:29:17 -0700 (MST) 12, 26 -- Date: Thu, 23 Feb 2006 08:29:17 -0700 12, 26 -- From: Zhenquan Liu {Zhenquan.Liu-at-asu.edu} 12, 26 -- Subject: TEM jobs 12, 26 -- To: Microscopy-at-microscopy.com 12, 26 -- Message-id: {60D5D23B9F20DF4C8DC3991979F9094A78D0DF-at-EX03.asurite.ad.asu.edu} 12, 26 -- MIME-version: 1.0 12, 26 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 12, 26 -- Content-type: text/plain; charset=us-ascii 12, 26 -- Thread-Topic: TEM jobs 12, 26 -- Thread-Index: AcY4jeeiwKNTEODRTQqsyzg0oBZSxQ== 12, 26 -- Content-class: urn:content-classes:message 12, 26 -- X-MS-Has-Attach: 12, 26 -- X-MS-TNEF-Correlator: 12, 26 -- Content-Transfer-Encoding: 8bit 12, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1NFThIY010225 ==============================End of - Headers==============================
Not sure about a pen, but we use a diamond scribe (Zeiss) mounted to the scope's turret to mark individual cells on a glass coverslip. This can go down to some 0.2 mm diameter, but, of course, leaves a scratch that cannot be removed. If you are interested, let me know and I will check the part no etc. (It was a little bit of pain to get it from them.)
m.
sirapa-at-optonline.net wrote:
--| --|Title-Subject: [Filtered] Glass Slide marker --| --|Question: I am looking for an ink based glass slide marker that can be mounted to the nosepiece of an optical microscope and mark a viewed area for later observation. I found one from Ernest Fullam that marks a 3mm diamter circle but need to mark a much smaller area. Any suggestions? --| --|--------------------------------------------------------------------------- --|
==============================Original Headers============================== 7, 19 -- From M_Jarnik-at-fccc.edu Thu Feb 23 09:49:43 2006 7, 19 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 7, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NFngKS019961 7, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 09:49:43 -0600 7, 19 -- Received: from fccc.edu (emf4.fccc.edu [131.249.9.170]) 7, 19 -- by azah.fccc.edu (8.12.11/8.12.11) with ESMTP id k1NFngaO022470 7, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 10:49:42 -0500 (EST) 7, 19 -- Message-ID: {43FDD996.3070402-at-fccc.edu} 7, 19 -- Date: Thu, 23 Feb 2006 10:49:42 -0500 7, 19 -- From: Michal Jarnik {M_Jarnik-at-fccc.edu} 7, 19 -- Reply-To: M_Jarnik-at-fccc.edu 7, 19 -- Organization: Fox Chase Cancer Center 7, 19 -- User-Agent: Mozilla/5.0 (Macintosh; U; PPC Mac OS X Mach-O; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 7, 19 -- X-Accept-Language: en,cs 7, 19 -- MIME-Version: 1.0 7, 19 -- To: Microscopy {Microscopy-at-microscopy.com} 7, 19 -- Subject: Re: Glass Slide marker 7, 19 -- Content-Type: text/plain; charset=us-ascii; format=flowed 7, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
You can use an 'England finder' glass slide. You remove the specimen slide and put in the England finder slide. It has exact co-ordinates etched onto the slide surface (you should use cross-hairs within the eyepiece for greatest accuracy). The process is reversed to go back to exactly the same place. Agar Scientific sell one over here for around Ł100 without the crosshair eyepiece graticule (item L4076 ). They also sell various 'Graticule Ltd' crosshair (eg cross gauge or cross micrometer) eyepiece graticules to drop into the eyepiece. See www.agarscientific.com.
As you say the printing objective 'red ring' systems or the XY stage micrometer scale if fitted but these are rather coarse.
regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {M_Jarnik-at-fccc.edu} To: {keith.morris-at-ucl.ac.uk} Sent: Thursday, February 23, 2006 3:54 PM
On Feb 23, 2006, at 7:04 AM, tonygr-at-MIT.EDU wrote:
} The sense I have is that the main problem comes about when the fresh } film is removed from the packet, and individual sheets are taken off } the stack. The discharging occurs between the surface of the } emulsion and the substrate of the next sheet of film. This seems to } me to be unconnected with the grounding of the user, but a problem } with the dryness of the film itself, and its highly non-conductive } surface. The film is stored in air, and the problem is much less } severe in the summer. We load the film in the film carriers before } dessiccation, so the handling afterwards is minimized. I may try } keeping the opened packets of film in a cabinet with a dish of water, } to keep the humidity of the film up. } Dear Tony, That is my sense also. The problem with keeping the film moist is that it will take much longer to pump down the camera, and the column vacuum may degrade when wet film is transported into position for exposure. If you only shoot one load of film per day and can pump on the camera overnight, this will not be too great a consideration, but otherwise waiting for a suitable camera vacuum will be very inconvenient. 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 Feb 23 12:06:30 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NI6UCB008524 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 12:06:30 -0600 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 6DEF935D89 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 10:06:29 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 69E3235D66 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 10:06:28 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602231504.k1NF4oiC032400-at-ns.microscopy.com} 4, 22 -- References: {200602231504.k1NF4oiC032400-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {eb3fc3a1806063103a4657d1a797ae5f-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] TEM film electrostatic discharging 4, 22 -- Date: Thu, 23 Feb 2006 10:14:36 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
One of my coworkers looked at the images and came up with a better suggestion than my previous one, in my humble opinion. She thought the objects resembled spores and I tend to agree with her after revisiting the images. I seem to recall that many spores, like some seeds, can survive passage through the digestive system, and this would better explain the apparent absence of a Si peak. The holes might be germination pores?
One of the images does appear to have a diatom in the background, though.
If this turns out to be correct, the credit goes to Melainia McClain, one of the newest and most enthusiastic entrants into the field of electron microscopy.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 24 -- From TindallR-at-missouri.edu Thu Feb 23 12:38:17 2006 10, 24 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 10, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NIcH0Y018311 10, 24 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 12:38:17 -0600 10, 24 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 24 -- Thu, 23 Feb 2006 12:38:17 -0600 10, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 24 -- Content-class: urn:content-classes:message 10, 24 -- MIME-Version: 1.0 10, 24 -- Content-Type: text/plain; 10, 24 -- charset="US-ASCII" 10, 24 -- Subject: Mystery objects 10, 24 -- Date: Thu, 23 Feb 2006 12:38:15 -0600 10, 24 -- Message-ID: {BA876152E8653240BE8572E897083EE701849DB9-at-UM-EMAIL09.um.umsystem.edu} 10, 24 -- X-MS-Has-Attach: 10, 24 -- X-MS-TNEF-Correlator: 10, 24 -- Thread-Topic: Mystery objects 10, 24 -- Thread-Index: AcY4qE+TWuHpDwfRTY+h+Wrc8cs5eg== 10, 24 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 24 -- To: {jehrman-at-mta.ca} 10, 24 -- Cc: {microscopy-at-microscopy.com} 10, 24 -- X-OriginalArrivalTime: 23 Feb 2006 18:38:17.0269 (UTC) FILETIME=[4FE4BE50:01C638A8] 10, 24 -- Content-Transfer-Encoding: 8bit 10, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1NIcH0Y018311 ==============================End of - Headers==============================
You guys are amazing! I have located one. Most of the people replied has the older model 502 which is quite different from the 502A especially with the autopumping option. There may be some parts such as those with the filament holders missing but I am certainly much closer to getting the machine back to working condition. Thanks for all your help.
-- Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519) The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
} ---------------------------------------------------------------------------- } } I wonder if anyone has a manual for the Denton DV-502A vacuum evaporator } that I can borrow. Ours was lost during moving to storage and now I have } to set it up again. It would be nice to make sure I set it up correctly. } Denton can supply the manual for $250 so I am looking at other less } expensive ways first. Thanks a lot!
==============================Original Headers============================== 3, 21 -- From wpchan-at-u.washington.edu Thu Feb 23 13:19:28 2006 3, 21 -- Received: from mxout2.cac.washington.edu (mxout2.cac.washington.edu [140.142.33.4]) 3, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NJJQxI028137 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 13:19:27 -0600 3, 21 -- Received: from homer23.u.washington.edu (homer23.u.washington.edu [140.142.12.141]) 3, 21 -- by mxout2.cac.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k1NJJPYg030432 3, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 11:19:25 -0800 3, 21 -- Received: from localhost (wpchan-at-localhost) 3, 21 -- by homer23.u.washington.edu (8.13.5+UW05.10/8.13.5+Submit) with ESMTP id k1NJJP8b005316 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 11:19:25 -0800 3, 21 -- Date: Thu, 23 Feb 2006 11:19:25 -0800 (PST) 3, 21 -- From: "W. Chan" {wpchan-at-u.washington.edu} 3, 21 -- To: microscopy-at-microscopy.com 3, 21 -- Subject: RE: [Microscopy] manual for Denton DV-502A 3, 21 -- In-Reply-To: {FA26BEF1EA775E4584FB34B91E14A1C404DBC6E2-at-SCHMLVEM01.e2k.ad.ge.com} 3, 21 -- Message-ID: {Pine.LNX.4.64.0602231111070.1855-at-homer23.u.washington.edu} 3, 21 -- References: {FA26BEF1EA775E4584FB34B91E14A1C404DBC6E2-at-SCHMLVEM01.e2k.ad.ge.com} 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 3, 21 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0, __STOCK_PHRASE_7 0' ==============================End of - Headers==============================
Wow! I should post weird stuff more often. Finally, a day where interesting email outnumbers spam! As requested by a number of respondents, I've compiled your responses which appear below. In case you just wandered in, we're discussing the objects found in wallaby droppings - images posted at:
http://www.mta.ca/dmf/download/ehrman/mystery.htm
I also have a number of clarifications and comments inspired by your responses:
1. No, this is not a joke!
2. It's highly unlikely that these are some artifact associated with the polycarbonate filter. We processed about 62 samples in between other types of samples, using filters from the same box. Only the wallaby samples had these objects on them, and I've never seen anything like this on any filter I've used. While the samples were treated with 5% KOH (for less than 30 minutes, mainly to dissolve and disperse mucous) the filters actually saw a much less concentrated solution - typically 5 - 40 drops of sieved droppings were added to the filter stack filled with 15 ml dH2O. There was absolutely no evidence of any compromising of the filters anywhere on the specimens. I would be quite surprised if anything happened to the filters - they survive virtually unchanged with boiling for 60 minutes in a 50:50 mixture of concentrated sulfuric and nitric acids (used for cleaning diatoms).
3. We also don't think that they are diatoms, fungus spores, fern spores, or pollen. We work mainly with diatoms here, and we've never seen anything remotely similar. The material was being examined for mushroom and truffle spores, which were very abundant and generally appeared unmolested in comparison with undigested spores. There certainly were diatoms in the samples (probably from drinking water) but they appeared as expected. Also many phytoliths, trematode, cestode and nematode eggs and bits of plant structures that we've seen before in other contexts and could at least be somewhat certain of a likely identity. These mysterious structures, however, I haven't run across in almost 30 years of looking at a wide rang of really strange samples - wallaby poo is probably only somewhere in the middle!
4. These objects are definitely not siliceous, at least to the extent diatoms are. With this amount of gold coating, a reasonably robust diatom with a frustule as thick as these objects are would generate a very substantial Si peak (trust me on this one). As I mentioned before, they were easily torn up by a 10-15 kV beam with a spot size sufficient for collection of EDS spectra. The diatoms, phytoliths, calcium oxalate crystals, calcium carbonates, etc. in the sample were quite resistant to the same beam and peak heights from the expected elements were at least the same order of magnitude as the Au-M alpha line.
So, looks like it's still a mystery. But thanks for all your input, and if I learn anything new, I'll let you know.
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
-- My vote is that it is a micro-alien's dice. I use the plural term because of the old admonition: "Never say die!" -- No, but it has been some time since a microscopy list post has made me laugh out loud. Thank you... -- They look an awful lot like the Nuclepore filter in the background. Hole size, Smooth walls on the holes etc. -- How bizarre! Is it possible that they are polycarbonate contaminants on your polycarbonate filter? The pores look to be the same size and I'd imagine the spectrum would be similar. -- If I think back to my botany training at the University of Guelph, I'd think they were diatoms except I looked at your EDS and didn't see a peak for silica. Good luck! -- Very cool photos. Interesting wallaby you got there. I have no references, but it suggests a sieve plate from vegetation, at least that my uneducated guess. please put me in the pool of guesses and let us know who's right when you figure it out!! -- I wonder if they're the remains of thick-walled plant cells, the holes would be the remains of pitfields where the wall is thinner. Some cells have walls that are very resistant to digestion. They do seem amazingly clean and smooth for plant cells though. -- Can you distinguish the objects with light microscopy? If they are modified plant cells they should stain blue (cellulose) in a Zinc Chloride - Iodine solution and light up with polarized light. -- my very uneducated guess (I mostly do TEM of cells & tissues) would be some sort of pollen grain or even diatoms (in the soil..is it chalky?) that got ingested up when your furry friend ate. I'd be very curious to see what it is, please post a summary or solution when you arrive at one! -- Looks like it might be some sort of polystyrene or other foam that has been weathered, eaten and passed. Not sure though. Maybe compare the pore size with known plastic foams. Good luck! -- The 5% KOH is relevant, as it's a good way to clean chitinous structures and maybe heavy-walled botanical things. I'd wager on either digested pollen or spores from ferns or fungi. Can you ID what plants the critters were eating, and whether the plants were in bloom or producing spores? -- This is a total guess, but they could be pollen. You can get some very interesting structures with pollen. -- .... nice pictures, reminds me of pollen grains (size, form, smoothness of surface). will you publish the outcome of your riddle in the discussion group? -- being no specialist (neither in SEM-image interpretation nor SEM specimen preparation but remembering the chapter: } What izz' it { in Ultrastructural Pathology or the Chapter } What is it ? { in the Proceedings of the RMS) I only can guess:
Nano-Soccer World Championship 2006 {: some samples of the "round leather" (as we call the } foot {-"balls" it here in Europe)...type "economy class" with holes in it for proper } aerodynamics {........no, seriousely, I do not have any invulnerable idea what those structures might be, except some kind of "vegetable" remnants including specialized perhaps symbiontic algae with a hard shell intermingled with the vegetable food wallabys perhaps are taking up........assuming that the only elements you found (cf. spectrum) are C and O (Au from sputtering?).......what about } Si { ? Perhaps you should forward your images to Phytotomists working in the field of algae. -- have you considered pollen? -- I showed your images to a colleague who does SEM of pollen, and she thinks that the pores are too irregular and the structure too smooth to be pollen remnants. She routinely uses acid digestion to get rid of coatings on the pollen, and her SEMs show lots of microstructure, and layering in the pore wall, neither of which your samples show. So, I guess pollen is out. She was skeptical of spores (fern or fungus) for many of the same reasons (and size for fungal spores -- yours are likely too big). So, I'm puzzled. -- Einstein said 'God does not throw dice', but apparently wallabies do. Forget my facetious* sight-unseen 'diamonds' thesis: these boogers are organic. They look 'waxy', not crystaline. Any chance of zooming in for microstructure? I'm betting they're laminate, perhaps cross-laminate. The centre hole in 0003 seems to be imbricated in section. I can't dispell the impression that the holes occur in PAIRs, especially 0003. Little hole going in, fatter hole coming out (after a meal?). I am forwarding this to 'Doctor Stats', in case he would like to take up the mathematical challenge of falsifying my Pairing Hypothesis. The carbon makeup makes me think of lignin structures. Perhaps a xeric plant has stowed away water or oils in lignin-lined vacuoles (and something vermiform has tunneled in, and out again?). -- I suspect that this is some silicaceous remnant of something in the wallaby's diet--something like diatom casts, plant opal phytoliths, or some related thing. Like Wolfgang, I assume (?) that the Au comes from sputtering and is simply overwhelming a silicon signal in the background. I have never personally seen a phytolith quite like these, but one of the preeminent phytolith specialists,is on our campus and I will forward your link to her, if you would resend it to me. I accidentally deleted the message yesterday. I love stuff like this, especially when people ask me "What kinds of things do EM people look at?". I can now add wallaby poop to the distinguished and growing list. -- Digested diatoms or pollen grains? -- Looks like you need to look at the specimen without Au coating, or with very minimal Au coating, so that you can determine what is the composition of the object of interest. Right now your data indicates that it is carbonaceous, with minimal Si, maybe like a primative seed-coat or a spore-coat. The structures appear quite regular from the images provided. The holes appear to be etch pits. Apparently you have sputtered lots of Au on the specimen. Your EDS spectrum indicates no presence of any elements with major x-ray lines above 2.5 KeV. Therefore you may Reduce, eliminate or change the coating material, and reduce the accelerating voltage to ~5 KeV to reduce charging, and then see (qualitatively) what the composition is of these nano-footballs. Secondly, I would compare the size and shape of these objects to spore-like structures from the plants, mosses and fungi in the potential Wallaby diet. -- Haven't got the foggiest what they are but I could be convinced that the holes do not occur at random. -- Is it a joke? It seems to me like spherical pieces or remnants of polycarbonate filter. The "pore" size is close to the size of pores in supporting filter membrane and the x-ray spectrum contains only C and O "and Au from coating". -- One of my coworkers looked at the images and came up with a better suggestion than my previous one, in my humble opinion. She thought the objects resembled spores and I tend to agree with her after revisiting the images. I seem to recall that many spores, like some seeds, can survive passage through the digestive system, and this would better explain the apparent absence of a Si peak. The holes might be germination pores? One of the images does appear to have a diatom in the background, though. If this turns out to be correct, the credit goes to Melainia McClain, one of the newest and most enthusiastic entrants into the field of electron microscopy.
==============================Original Headers============================== 16, 17 -- From jehrman-at-mta.ca Thu Feb 23 14:11:58 2006 16, 17 -- Received: from mailserv.mta.ca (mailserv.mta.ca [138.73.1.1]) 16, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NKBvxV005797 16, 17 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 23 Feb 2006 14:11:57 -0600 16, 17 -- Received: from host-22-245.mta.ca ([138.73.22.245]) 16, 17 -- by mailserv.mta.ca with esmtp (Exim 4.52) 16, 17 -- id 1FCMod-0006DB-0p; Thu, 23 Feb 2006 16:11:56 -0400 16, 17 -- Message-ID: {43FE1704.10002-at-mta.ca} 16, 17 -- Date: Thu, 23 Feb 2006 16:11:48 -0400 16, 17 -- From: "James M. Ehrman" {jehrman-at-mta.ca} 16, 17 -- Organization: Mount Allison University 16, 17 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 16, 17 -- MIME-Version: 1.0 16, 17 -- To: Microscopy Listserv {Microscopy-at-MSA.Microscopy.com} 16, 17 -- Subject: mystery object revisited 16, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 16, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
When I was loading Kodak 4489 (or SO163) film into film boxes for my Phillips 300 I always (1) wore non-powdered latex gloves and (2) always put a fresh packet of film wrapped in the original foil package but sliced open with a razor blade into the desiccator on top of the reserve loaded film box that was kept ready to use, thus the film was already outgassed before loading into the film holders. The old Phillips only held 16 plates at a time so I changed film a lot. I never had a problem with electrostatic discharging. I admit that I had lots of other problems but that's another story.
On Feb 23, 2006, at 7:04 AM, tonygr-at-MIT.EDU wrote: The sense I have is that the main problem comes about when the fresh film is removed from the packet, and individual sheets are taken off the stack. The discharging occurs between the surface of the emulsion and the substrate of the next sheet of film. This seems to me to be unconnected with the grounding of the user, but a problem with the dryness of the film itself, and its highly non-conductive surface. The film is stored in air, and the problem is much less severe in the summer. We load the film in the film carriers before dessiccation, so the handling afterwards is minimized. I may try keeping the opened packets of film in a cabinet with a dish of water, to keep the humidity of the film up.
Dean Abel Biological Sciences University of Iowa Iowa City IA 52242-1324
==============================Original Headers============================== 5, 17 -- From dean-abel-at-uiowa.edu Thu Feb 23 14:38:31 2006 5, 17 -- Received: from day.its.uiowa.edu (day.its.uiowa.edu [128.255.56.107]) 5, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NKcVt7015428 5, 17 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 14:38:31 -0600 5, 17 -- Received: from abel01.uiowa.edu (dhcp80ff2538.dynamic.uiowa.edu [128.255.37.56]) 5, 17 -- by day.its.uiowa.edu (8.12.10/8.12.9/ns-mx-1.17) with ESMTP id k1NKcRN1076546; 5, 17 -- Thu, 23 Feb 2006 14:38:29 -0600 5, 17 -- Message-Id: {6.2.1.2.2.20060223141815.01d42cc0-at-blue.weeg.uiowa.edu} 5, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 5, 17 -- Date: Thu, 23 Feb 2006 14:28:54 -0600 5, 17 -- To: microscopy-at-microscopy.com, tivol-at-caltech.edu; 5, 17 -- From: Dean Abel {dean-abel-at-uiowa.edu} 5, 17 -- Subject: Re: [Microscopy] TEM film electrostatic discharging 5, 17 -- In-Reply-To: {200602231808.k1NI8wTG011830-at-ns.microscopy.com} 5, 17 -- References: {200602231808.k1NI8wTG011830-at-ns.microscopy.com} 5, 17 -- Mime-Version: 1.0 5, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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==============================Original Headers============================== 3, 17 -- From larry-at-cymru.freewire.co.uk Thu Feb 23 15:04:15 2006 3, 17 -- Received: from get.freewire.net (get.freewire.net [195.184.229.250]) 3, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NL4Ee1025233 3, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 23 Feb 2006 15:04:14 -0600 3, 17 -- Received: from [217.154.255.187] ([217.154.255.187]) 3, 17 -- by get.freewire.net (8.11.6/8.11.6) with ESMTP id k1NL49W23138; 3, 17 -- Thu, 23 Feb 2006 21:04:09 GMT 3, 17 -- Mime-Version: 1.0 3, 17 -- Message-Id: {p06210203c023d02f05b2-at-[217.154.253.116]} 3, 17 -- In-Reply-To: {200602222345.k1MNjAg4024882-at-ns.microscopy.com} 3, 17 -- References: {200602222345.k1MNjAg4024882-at-ns.microscopy.com} 3, 17 -- Date: Thu, 23 Feb 2006 20:51:14 +0000 3, 17 -- To: walck-at-southbaytech.com, Microscopy-at-MSA.Microscopy.Com 3, 17 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 3, 17 -- Subject: Re: [Microscopy] ROHS and European microscopists who are 3, 17 -- fishermen (or fisherwomen) 3, 17 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
A colleague is looking for some histology equipment. Below is her wish list. You can contact her at bcarlsward-at-flmnh.ufl.edu
Here’s a list of the equipment that I’m looking for:
* Rotary microtome for paraffin work * Sliding microtome for unembedded specimens (I don’t think anyone will have a spare of either microtome, but it doesn’t hurt to ask) * Long and short microtome knives (for the sliding and rotary microtomes), nicked blades are OK * Knife sharpener (for long and short blades) * Slide warming tables (nothing fancy, I don’t even need a temperature gauge) * Paraffin oven (again, nothing fancy, just something I can melt paraffin in)
-- Karen L. Kelley ICBR Electron Microscopy Manager University of Florida ICBR Electron Microscopy Core Lab Building 747, Room 214 Box 118525 Gainesville Florida Lab: 352-392-1184 fax: 352-846-0251 Email: klk-at-biotech.ufl.edu Southeastern Microscopy Society Treasurer http://www.biotech.ufl.edu/EM/
==============================Original Headers============================== 7, 21 -- From klk-at-biotech.ufl.edu Thu Feb 23 15:14:07 2006 7, 21 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NLE74R002301 7, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 15:14:07 -0600 7, 21 -- Received: from [128.227.60.41] (empc41719.dhcp.clas.ufl.edu [128.227.60.41]) 7, 21 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k1NLE5Be4423872 7, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 16:14:06 -0500 7, 21 -- Message-ID: {43FE259D.7000405-at-biotech.ufl.edu} 7, 21 -- Date: Thu, 23 Feb 2006 16:14:05 -0500 7, 21 -- From: Karen Kelley {klk-at-biotech.ufl.edu} 7, 21 -- User-Agent: Mozilla Thunderbird 1.0.6 (Windows/20050716) 7, 21 -- X-Accept-Language: en-us, en 7, 21 -- MIME-Version: 1.0 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- Subject: looking for equipment 7, 21 -- Content-Type: text/plain; charset=windows-1252; format=flowed 7, 21 -- Content-Transfer-Encoding: 8bit 7, 21 -- X-Spam-Status: hits=0.001, required=5, tests=BAYES_50 7, 21 -- X-UFL-Spam-Status: hits=0.001, required=5, tests=BAYES_50 7, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 7, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Does anyone know whether or not waving a magic wand, laden with ethylene dichloride, over wrinkled sections will destroy the tissue's antigenicity for post-embedding immunogold labeling? I'd love to relax out the wrinkles, but I have been hesitating to use the ethylene dichloride trick for fear of altering the tissue's antigenicity. The tissue is embedded in Epon.
Has anyone tried this before?
Thanks for any ideas you might have on this.
Dotty Sorenson Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 6, 16 -- From dsoren-at-umich.edu Thu Feb 23 15:57:23 2006 6, 16 -- Received: from playinggod.mr.itd.umich.edu (playinggod.mr.itd.umich.edu [141.211.14.79]) 6, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NLvNln012420 6, 16 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 15:57:23 -0600 6, 16 -- Received: from [141.214.170.47] (host-47.subnet-170.med.umich.edu [141.214.170.47]) 6, 16 -- by playinggod.mr.itd.umich.edu (smtp) with ESMTP id k1NLvNt5006529 6, 16 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 16:57:23 -0500 6, 16 -- Mime-Version: 1.0 (Apple Message framework v746.2) 6, 16 -- Content-Transfer-Encoding: 7bit 6, 16 -- Message-Id: {1BC161D6-7469-491A-9E1E-9FC8B3F85400-at-umich.edu} 6, 16 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 16 -- To: microscopy-at-msa.microscopy.com 6, 16 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 6, 16 -- Subject: Wrinkles and ethylene dichloride 6, 16 -- Date: Thu, 23 Feb 2006 16:55:37 -0500 6, 16 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
Why not run your own test with your material, your antibodies, fixation protocol, processing protocol, etc. etc. ?
Geoff
dsoren-at-umich.edu wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 8, 31 -- From mcauliff-at-umdnj.edu Thu Feb 23 16:10:12 2006 8, 31 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NMACNT023263 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 16:10:12 -0600 8, 31 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 8, 31 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id ADD3E1C32BC 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 17:10:11 -0500 (EST) 8, 31 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 8, 31 -- by zix01.umdnj.edu (Proprietary) with ESMTP id D72FAA7B4C 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 17:10:08 -0500 (EST) 8, 31 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 31 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 31 -- id {0IV500K01UABBN-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 31 -- for microscopy-at-msa.microscopy.com; Thu, 23 Feb 2006 17:10:08 -0500 (EST) 8, 31 -- Received: from [127.0.0.1] ([10.138.2.240]) 8, 31 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 31 -- 2004)) with ESMTP id {0IV500HMPUWVSJ-at-Polaris.umdnj.edu} ; Thu, 8, 31 -- 23 Feb 2006 17:10:07 -0500 (EST) 8, 31 -- Date: Thu, 23 Feb 2006 17:09:22 -0500 8, 31 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 31 -- Subject: Re: [Microscopy] Wrinkles and ethylene dichloride 8, 31 -- In-reply-to: {200602232158.k1NLwc5Q014453-at-ns.microscopy.com} 8, 31 -- To: dsoren-at-umich.edu, MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 31 -- Message-id: {43FE3292.6010200-at-umdnj.edu} 8, 31 -- MIME-version: 1.0 8, 31 -- Content-type: text/plain; format=flowed; charset=us-ascii 8, 31 -- Content-transfer-encoding: 7BIT 8, 31 -- X-Accept-Language: en-us, en 8, 31 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 31 -- Gecko/20040804 Netscape/7.2 (ax) 8, 31 -- References: {200602232158.k1NLwc5Q014453-at-ns.microscopy.com} ==============================End of - Headers==============================
Why breathe ethylene dichloride fumes? I use a heat pen (mine is from Ted Pella but I believe others brands are available). It works better than any organic solvent I have ever used. I find no effect on antigenicity. Tom
At 03:58 PM 02/23/06, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
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Title-Subject: [Filtered] 2 photon induced DNA damage????
Question: Dear Confocalists,
There's a wonderful Nature Genetics paper describing TRF2 association with ds breaks (Volume 37, number 2, Feb 2005 Bradshaw, Stavropoulos, Meyn). They irradiate using 390nm laser (Cell Robotics laser scissors) to induce DNA damage.
We have an investigator here who would like to do something similar to this. We have a Zeiss 510 NLO with Argon, 543Hene, 633 HeNe and Chameleon 2p lasers.
My naive question is: can we induce DNA damage with our system?? I'm thinking perhaps of using the 2p laser tuned to around 780nm and the Edit Bleach function using a thin line for the ROI in order to "cut" through a nucleus. Or perhaps just a line scan would work??? But, of course, this begs the question: would the 2 photon effect have the same effect as UV light on DNA??
We're brand new to 2p microscopy, so would appreciate ANY insight or recommendations.
I am not sure what you are asking. The real sizes of what? Bill's reply addresses calibration of negative mags versus gratings, etc. I read your question as, "I know my negative mag is 80,000X from my TEM 'read out' and the carbon grating replica calibration. There is no micron marker on the negative anymore and so how long is one micron on that negative (or on a print I make)?" 80 mms = 1 µm.
This calculation works on SEM prints, TEM prints, optical micrographs, and negatives where you know the magnification.
********************* GENERAL MAG CALCULATION RULE: Calculate the known magnification in KX. That numerical KX value in millimeters is the length of one micrometer (µm) on your print, negative, or whatever. ********************* For example, you have a final print mag of 125,000X. If you draw a line 125 millimeters in length, that will be the length of one micron on that print. For a tenth of a micron (0.1 µm), just use 12.5 mm. This rule is simple to remember and it works on any photographic media for which you know the magnification.
You can reverse the procedure to calculate the magnification on a journal article micrograph without a stated magnification but with a micron scale. Calculate the length that it would take to make one micron with the scale marker shown on the micrograph. That number in millimeters, is the KX value of the magnification of the photograph in the article.
A bit more: A (no arrowheads) line scale marker can be made with Photoshop on electronically scanned images of negatives. See the reference below for a detailed discussion. Basically, select the line tool (/). Hold down the shift key. With the line option window tab set to a width of 5-10 and the preferences set to CM units in the PS preferences, mouse drag a horizontal line to the length that you need. Look in the info window to read the current line length. The length displayed will be in CM, not MMs, and so you can get confused at times. Release the mouse button to make the proper line length as shown in the info window / tab.
To make the micron, µ, character; hold down the ALT key. Type 0181 on the NUM PAD using those keys. DO NOT use the regular number keys at the top of the keyboard. Release the ALT key and the micron symbol, µ, will appear. This works on any PC clone and in any program running on a windows platform, even email clients. Woolley, below, also claims a method for Macs on ISO-Latin-1 characters.
10 µm |----------|
If the serial numbering and micron marking are both broken, use a Sanford Sharpie® extra fine marking pen to write waterproof and D-19 developer-proof numbers on your TEM film before use. They work for manually drawing line length markings on prints too.
I hope this helps you out.
Paul Beauregard
Notes: An Optical Microscopy example. The mag is 500X. That's 0.5 KX. So one micron is 0.5 mm. That is too small of a length to draw. So multiply by ten and use 5 mms. Label that 5 mm length as 10 microns.
Ref: Microscopy.com archive. On the home page in Microscopy.com at the bottom left, click on "Search the archives" Use author Beauregard and look in "all of 2002" or just Feb, 2002. Look for: Subject: Straight parallel micron bars in Photoshop®. Feb 4, 2002.
At 04:32 PM 2/18/06 -0600, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 18, 27 -- From beaurega-at-westol.com Thu Feb 23 19:34:04 2006 18, 27 -- Received: from smtp-gateway-2.winbeam.com (smtp-gateway-2.winbeam.com [64.84.97.67]) 18, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1O1Y3Pu008818 18, 27 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 19:34:04 -0600 18, 27 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 18, 27 -- by smtp-gateway-2.winbeam.com (8.13.1/8.12.8) with SMTP id k1O1Y0HY003460 18, 27 -- for {microscopy-at-microscopy.com} ; Thu, 23 Feb 2006 20:34:00 -0500 18, 27 -- Received: (qmail 9079 invoked by uid 89); 24 Feb 2006 01:33:58 -0000 18, 27 -- Received: from 064-084-106-036.gbg-07.modem.cust.winbeam.com (HELO millenium) (64.84.106.36) 18, 27 -- by mail.winbeam.com with SMTP; 24 Feb 2006 01:33:58 -0000 18, 27 -- Message-Id: {3.0.6.32.20060223173355.007b5d20-at-pop3.norton.antivirus} 18, 27 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 18, 27 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 18, 27 -- Date: Thu, 23 Feb 2006 17:33:55 -0800 18, 27 -- To: soleimanij-at-tbzmed.ac.ir, microscopy-at-microscopy.com 18, 27 -- From: Beaurega {beaurega-at-westol.com} 18, 27 -- Subject: Re: [Microscopy] viaWWW: scale bar 18, 27 -- In-Reply-To: {200602182232.k1IMWsoU028633-at-ns.microscopy.com} 18, 27 -- Mime-Version: 1.0 18, 27 -- Content-Type: text/plain; charset="iso-8859-1" 18, 27 -- X-Winbeam-MailScanner-Information: - Please contact Technical Support for more information 18, 27 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) 18, 27 -- X-Winbeam-MailScanner-SpamCheck: not spam, SpamAssassin (score=-1.44, 18, 27 -- required 6, autolearn=disabled, ALL_TRUSTED -1.44) 18, 27 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com 18, 27 -- Content-Transfer-Encoding: 8bit 18, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1O1Y3Pu008818 ==============================End of - Headers==============================
Further to the England finder, you can of course just buy pre-etched cover slips with all the XY co-ordinates eteched onto them, then the location of the bit of interest is always integrated into the specimen slide (unless the coverslip slides about in a temporary mount). You can also use a 'Graticule Ltd' crosshair eyepiece graticule for greater accuracy with complex tissues. See the cover slips at: http://www.proscitech.com.au/catalogue/g2.asp near the bottom of the page (available as round or square pre-etched coverslips)
You can also get these coverslips glued onto the base of a plastic petri dish to serve the same purpose with live cells on an inverted microscope. See http://www.mattek.com/ and http://www.glass-bottom-dishes.com/ specifically for Petri dishes (and multiwell plates). To quote "The gridded coverslips allow one to refer to specific cells and follow them over time. For instance, individual cells can be microinjected, returned to the incubator, and observed at multiple time points since each cell can be identified with a unique alpha-numeric coordinate in the dish. Glass bottom dishes containing Bellco Glass coverslips are available. Part numbers for standard gridded dishes are: P35G-2-14-C-GRID and P50G-2-14-F-GRID".
Additionally you can glue a couple of micrometer scales to the X and Y of the manual stage surface at the movment interface to get a rough guide to position - we had our workshop do this with a couple of cheap Leica DM1L microscopes (for multiwell plates mainly, although it will help find an area quickly even with pre-ecthed coverslips). They cut out bits of etched steel rules and glued them on and the results were very professional looking.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {M_Jarnik-at-fccc.edu} To: {keith.morris-at-ucl.ac.uk} Sent: Thursday, February 23, 2006 3:54 PM
Hello Listers
I am trying to do some ruthenium red staining of tb, and have so far been unsuccessful. My initial fix is 2.5% glut, 0.05% RR in 0.1M cacodylate bufffer. I also use 0.05% RR 1% Osmium in 0.1M Cac buffer. (I have also used Hepes)
I have used the ruthenium red in the buffer washes and and also used just plain buffer for the washes. I just seem to get globs of RR by itself in the block and very little to nothing staining the cell. Any staining that I have seen is really patchy.
Is there a trick I'm missing?
Thanks in advance.
Leslie
Leslie Cummins www.aecom.yu.edu/aif Analytical Imaging Facility 1300 Morris Park Ave Forchheimer 639 Bronx, NY 10461 718-430-3547
==============================Original Headers============================== 8, 21 -- From gunther-at-aecom.yu.edu Fri Feb 24 07:47:20 2006 8, 21 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 8, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1ODlKxN004524 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 24 Feb 2006 07:47:20 -0600 8, 21 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 8, 21 -- by mailgw.aecom.yu.edu (8.12.11/8.12.11) with SMTP id k1ODlK8G024966 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 24 Feb 2006 08:47:20 -0500 8, 21 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 8, 21 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006022408472011864 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 24 Feb 2006 08:47:20 -0500 8, 21 -- Received: from auc.aecom.yu.edu (auc.aif.aecom.yu.edu [129.98.31.132]) 8, 21 -- by post.aecom.yu.edu (Postfix) with ESMTP id 32F632873 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 24 Feb 2006 08:47:20 -0500 (EST) 8, 21 -- Message-Id: {6.2.1.2.0.20060224084022.01ce97c8-at-mailserver.aecom.yu.edu} 8, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 8, 21 -- Date: Fri, 24 Feb 2006 08:47:11 -0500 8, 21 -- To: Microscopy-at-Microscopy.Com 8, 21 -- From: Leslie Cummins {gunther-at-aecom.yu.edu} 8, 21 -- Subject: Ruthenium red staining 8, 21 -- Mime-Version: 1.0 8, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
A few, well one nice person actually, have expressed an interest in the England finder. In case it is of any interest to others, I'll post the home-page of Graticules ltd. as it has more details of their large range of graticules (or is it reticles), including pictures of the said 'England finder' (they are based in England). It's:
http://www.graticules.com/
They also make the calibration (stage micrometer) slides, and such like, that many of us use.
I'm always fascinated by the little things that eyepiece reticles can do from measuring distances and area to counting and 'finding' stuff down a microscope.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
Yes, but how about your liver? Are there changes in it from breathing the xylene fumes?
At 03:27 AM 02/24/06, you wrote:
} HI Philipst } } we are regularly useing xylene vapours for } immunoelectron microscopy and } got very good results and there is no change in } antigencity and no more wrinkle sections } } shrunali } --- phillipst-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 } } } ---------------------------------------------------------------------------- } } } } Why breathe ethylene dichloride fumes? I use a heat } } pen (mine is from Ted } } Pella but I believe others brands are available). It } } works better than any } } organic solvent I have ever used. I find no effect } } on antigenicity. Tom } } } } } } At 03:58 PM 02/23/06, you wrote: } } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The } } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } ---------------------------------------------------------------------------- } } } } } } Dear listers, } } } } } } Does anyone know whether or not waving a magic } } wand, laden with } } } ethylene dichloride, over wrinkled sections will } } destroy the tissue's } } } antigenicity for post-embedding immunogold } } labeling? I'd love to } } } relax out the wrinkles, but I have been hesitating } } to use the } } } ethylene dichloride trick for fear of altering the } } tissue's } } } antigenicity. The tissue is embedded in Epon. } } } } } } Has anyone tried this before? } } } } } } Thanks for any ideas you might have on this. } } } } } } Dotty Sorenson } } } Dorothy Sorenson } } } Microscopy and Image-analysis Laboratory } } } Department of Cell and Developmental Biology } } } University Of Michigan Medical School } } } 4643 Medical Science Building II } } } 1301 Catherine } } } Ann Arbor, MI 48109-0616 } } } (734)763-1170 } } } FAX (734)763-1166 } } } } } } } } } ==============================Original } } Headers============================== } } } 6, 16 -- From dsoren-at-umich.edu Thu Feb 23 15:57:23 } } 2006 } } } 6, 16 -- Received: from playinggod.mr.itd.umich.edu } } } } } (playinggod.mr.itd.umich.edu [141.211.14.79]) } } } 6, 16 -- by ns.microscopy.com } } (8.12.11/8.12.8) with ESMTP id } } } k1NLvNln012420 } } } 6, 16 -- for } } {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 } } } 15:57:23 -0600 } } } 6, 16 -- Received: from [141.214.170.47] } } (host-47.subnet-170.med.umich.edu } } } [141.214.170.47]) } } } 6, 16 -- by playinggod.mr.itd.umich.edu } } (smtp) with ESMTP id } } } k1NLvNt5006529 } } } 6, 16 -- for } } {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 } } } 16:57:23 -0500 } } } 6, 16 -- Mime-Version: 1.0 (Apple Message framework } } v746.2) } } } 6, 16 -- Content-Transfer-Encoding: 7bit } } } 6, 16 -- Message-Id: } } {1BC161D6-7469-491A-9E1E-9FC8B3F85400-at-umich.edu} } } } 6, 16 -- Content-Type: text/plain; } } charset=US-ASCII; delsp=yes; format=flowed } } } 6, 16 -- To: microscopy-at-msa.microscopy.com } } } 6, 16 -- From: Dorothy Sorenson {dsoren-at-umich.edu} } } } 6, 16 -- Subject: Wrinkles and ethylene dichloride } } } 6, 16 -- Date: Thu, 23 Feb 2006 16:55:37 -0500 } } } 6, 16 -- X-Mailer: Apple Mail (2.746.2) } } } ==============================End of - } } Headers============================== } } } } Thomas E. Phillips, PhD } } Professor of Biological Sciences } } Director, Molecular Cytology Core } } 2 Tucker Hall } } University of Missouri } } Columbia, MO 65211-7400 } } } } 573-882-4712 (office) } } 573-882-0123 (fax) } } PhillipsT-at-missouri.edu } } } } } } } } ==============================Original } } Headers============================== } } 10, 21 -- From PhillipsT-at-missouri.edu Thu Feb 23 } } 16:40:47 2006 } } 10, 21 -- Received: from um-exproto9.um.umsystem.edu } } (um-exproto9.um.umsystem.edu [207.160.151.49]) } } 10, 21 -- by ns.microscopy.com (8.12.11/8.12.8) } } with ESMTP id k1NMekcx008095 } } 10, 21 -- for {Microscopy-at-msa.microscopy.com} ; Thu, } } 23 Feb 2006 16:40:47 -0600 } } 10, 21 -- Received: from um-exproto9.um.umsystem.edu } } ([207.160.151.148]) by um-exproto9.um.umsystem.edu } } with Microsoft SMTPSVC(6.0.3790.1830); } } 10, 21 -- Thu, 23 Feb 2006 16:40:46 -0600 } } 10, 21 -- Received: from phillips-dell.missouri.edu } } ([128.206.81.132]) by um-exproto9.um.umsystem.edu } } over TLS secured channel with Microsoft } } SMTPSVC(6.0.3790.1830); } } 10, 21 -- Thu, 23 Feb 2006 16:40:45 -0600 } } 10, 21 -- Message-Id: } } } {6.0.0.22.2.20060223163915.03dbf510-at-pop.missouri.edu} } } 10, 21 -- X-Sender: phillipst-at-pop.missouri.edu } } 10, 21 -- X-Mailer: QUALCOMM Windows Eudora Version } } 6.0.0.22 } } 10, 21 -- Date: Thu, 23 Feb 2006 16:40:43 -0600 } } 10, 21 -- To: dsoren-at-umich.edu } } 10, 21 -- From: Tom Phillips } } {phillipst-at-missouri.edu} } } 10, 21 -- Subject: Re: [Microscopy] Wrinkles and } } ethylene dichloride } } 10, 21 -- Cc: Microscopy-at-msa.microscopy.com } } 10, 21 -- In-Reply-To: } } {200602232158.k1NLwPqF013990-at-ns.microscopy.com} } } 10, 21 -- References: } } {200602232158.k1NLwPqF013990-at-ns.microscopy.com} } } 10, 21 -- Mime-Version: 1.0 } } 10, 21 -- Content-Type: text/plain; } } charset="us-ascii"; format=flowed } } 10, 21 -- X-OriginalArrivalTime: 23 Feb 2006 } } 22:40:45.0398 (UTC) FILETIME=[2F410F60:01C638CA] } } ==============================End of - } } Headers============================== } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Storing opened film/plates in a humid chamber is a good idea as long as it is for a short period of time. A constant high humidy may encourage fungal growth on film. My wife solves hardening of brown sugar by putting a piece of bread in a plastic bag with those sugar for a day. I think you may try this trick with a piece of moist paper towel. We had encounterd static discharge when loading pre-dessicated 35 mm bulk film. The problem has been solved by storing it in atmospherice condition and dessicating the loaded "camera" instead.
Ann Fook Yang EM Unit/ Unite EM AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 yanga-at-agr.gc.ca Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: tivol-at-caltech.edu [mailto:tivol-at-caltech.edu] Sent: Thursday, February 23, 2006 1:08 PM To: Yang, Ann-Fook
On Feb 23, 2006, at 7:04 AM, tonygr-at-MIT.EDU wrote:
} The sense I have is that the main problem comes about when the fresh } film is removed from the packet, and individual sheets are taken off } the stack. The discharging occurs between the surface of the } emulsion and the substrate of the next sheet of film. This seems to } me to be unconnected with the grounding of the user, but a problem } with the dryness of the film itself, and its highly non-conductive } surface. The film is stored in air, and the problem is much less } severe in the summer. We load the film in the film carriers before } dessiccation, so the handling afterwards is minimized. I may try } keeping the opened packets of film in a cabinet with a dish of water, } to keep the humidity of the film up. } Dear Tony, That is my sense also. The problem with keeping the film moist is that it will take much longer to pump down the camera, and the column vacuum may degrade when wet film is transported into position for exposure. If you only shoot one load of film per day and can pump on the camera overnight, this will not be too great a consideration, but otherwise waiting for a suitable camera vacuum will be very inconvenient. 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 Feb 23 12:06:30 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1NI6UCB008524 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 12:06:30 -0600 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 6DEF935D89 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 10:06:29 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 69E3235D66 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Feb 2006 10:06:28 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602231504.k1NF4oiC032400-at-ns.microscopy.com} 4, 22 -- References: {200602231504.k1NF4oiC032400-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {eb3fc3a1806063103a4657d1a797ae5f-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] TEM film electrostatic discharging 4, 22 -- Date: Thu, 23 Feb 2006 10:14:36 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
==============================Original Headers============================== 14, 29 -- From YANGA-at-AGR.GC.CA Fri Feb 24 08:45:37 2006 14, 29 -- Received: from agroutsmtp1.agr.gc.ca (agroutsmtp1.agr.gc.ca [192.197.71.175]) 14, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1OEjbLo031659 14, 29 -- for {microscopy-at-MSA.Microscopy.com} ; Fri, 24 Feb 2006 08:45:37 -0600 14, 29 -- Received: from agrin1-old.agr.gc.ca (agrgate.agr.ca [192.197.71.189]) 14, 29 -- by agroutsmtp1.agr.gc.ca (8.12.8/8.12.8) with ESMTP id k1OEjYHp029265; 14, 29 -- Fri, 24 Feb 2006 09:45:34 -0500 14, 29 -- Received: from onncrxcn1.AGR.GC.CA ([10.117.15.130]) 14, 29 -- by agrin1-old.agr.gc.ca (8.12.8/8.12.8) with ESMTP id k1OElUle024660; 14, 29 -- Fri, 24 Feb 2006 09:47:30 -0500 14, 29 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn1.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 14, 29 -- Fri, 24 Feb 2006 09:45:29 -0500 14, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 14, 29 -- Content-class: urn:content-classes:message 14, 29 -- MIME-Version: 1.0 14, 29 -- Content-Type: text/plain; 14, 29 -- charset="iso-8859-1" 14, 29 -- Subject: RE: [Microscopy] Re: TEM film electrostatic discharging 14, 29 -- Date: Fri, 24 Feb 2006 09:45:28 -0500 14, 29 -- Message-ID: {E035A9C87303AE4AB9BA10FD8324DFB102433485-at-onncrxms3.agr.gc.ca} 14, 29 -- X-MS-Has-Attach: 14, 29 -- X-MS-TNEF-Correlator: 14, 29 -- Thread-Topic: [Microscopy] Re: TEM film electrostatic discharging 14, 29 -- thread-index: AcY4pCTySE2l2xQGTAS3IWvmo9pOrAAqRR9w 14, 29 -- From: "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 14, 29 -- To: {tivol-at-caltech.edu} , {microscopy-at-MSA.Microscopy.com} 14, 29 -- X-OriginalArrivalTime: 24 Feb 2006 14:45:29.0103 (UTC) FILETIME=[F4A181F0:01C63950] 14, 29 -- Content-Transfer-Encoding: 8bit 14, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1OEjbLo031659 ==============================End of - Headers==============================
Have you considered the possibiity that these objects are partially degraded starch granules? This would explain the absence of any silicium peak and the size is about correct. We send you offline some pictures of partially degraded corn starch (sorry, we cannot put them in a WebPages just now). It gets a somehow similar sponge aspect. If the pores are an effect of degradation or incipient gelation they will have a random distribution, while in spores of diatoms quite likely the pattern is more regular.
Our pictures, which show polyhedral structures with holes, are corn starch granules after high pressure treatment, some of them in a basic medium which seems to increase the degradation degree.
Which kind of starch your samples may be we do not know but granule shape and size can help in identifying the plant origin. For the moment we only have found this pit or hole degradation at the surface of corn granules, while other plant species that we have checked follow a different behaviour.
We can suggest dying with iodine to see the blue colour in optical microscopy. Also the classical Maltese cross can be seen in starch granules when seen with polarised light.
Hoping that this can help,
Miriam Martino and Antonio Molina
CIDCA, Universidad Nacional de La Plata, Argentina Instituto del Frio (CSIC), Madrid, Spain
----- Original Message ----- X-from: {jehrman-at-mta.ca} To: {ifrm111-at-if.csic.es} Sent: Wednesday, February 22, 2006 10:02 PM
I used to wave a Q-tip dipped in chloroform over thin sections while they were still in the water bath, until that was declared too dangerous. Then I switched to a heat pen from Ted Pella Inc, which worked very well. I wasn't doing immuno, but the heat pen doesn't give off too much heat so it would probably be all right.
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Dear listers,
Does anyone know whether or not waving a magic wand, laden with ethylene dichloride, over wrinkled sections will destroy the tissue's antigenicity for post-embedding immunogold labeling? I'd love to relax out the wrinkles, but I have been hesitating to use the ethylene dichloride trick for fear of altering the tissue's antigenicity. The tissue is embedded in Epon.
Has anyone tried this before?
Thanks for any ideas you might have on this.
Dotty Sorenson Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 11, 26 -- From leswes-at-shaw.ca Fri Feb 24 10:38:32 2006 11, 26 -- Received: from pd2mo2so.prod.shaw.ca (shawidc-mo1.cg.shawcable.net [24.71.223.10]) 11, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1OGcWar021263 11, 26 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Feb 2006 10:38:32 -0600 11, 26 -- Received: from pd3mr1so.prod.shaw.ca 11, 26 -- (pd3mr1so-qfe3.prod.shaw.ca [10.0.141.177]) by l-daemon 11, 26 -- (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 15 2004)) 11, 26 -- with ESMTP id {0IV700AORA87KQ50-at-l-daemon} for Microscopy-at-microscopy.com; Fri, 11, 26 -- 24 Feb 2006 09:38:32 -0700 (MST) 11, 26 -- Received: from pn2ml2so.prod.shaw.ca ([10.0.121.146]) 11, 26 -- by pd3mr1so.prod.shaw.ca (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 11, 26 -- 15 2004)) with ESMTP id {0IV700C3IA87HB80-at-pd3mr1so.prod.shaw.ca} for 11, 26 -- Microscopy-at-microscopy.com; Fri, 24 Feb 2006 09:38:31 -0700 (MST) 11, 26 -- Received: from [24.84.44.216] by l-daemon 11, 26 -- (Sun ONE Messaging Server 6.0 HotFix 1.01 (built Mar 15 2004)) 11, 26 -- with ESMTP id {0IV700C0ZA87TR00-at-l-daemon} for Microscopy-at-microscopy.com; Fri, 11, 26 -- 24 Feb 2006 09:38:31 -0700 (MST) 11, 26 -- Date: Fri, 24 Feb 2006 08:39:56 -0800 11, 26 -- From: Lesley Weston {leswes-at-shaw.ca} 11, 26 -- Subject: Re: Wrinkles and ethylene dichloride 11, 26 -- To: Microscopy-at-microscopy.com 11, 26 -- Message-id: {C02476DB.4416%leswes-at-shaw.ca} 11, 26 -- MIME-version: 1.0 11, 26 -- Content-type: text/plain; charset=US-ASCII 11, 26 -- Content-transfer-encoding: 7bit 11, 26 -- User-Agent: Microsoft-Outlook-Express-Macintosh-Edition/5.0.6 ==============================End of - Headers==============================
Thanks for all the advice and opinions - I think the images Miriam Martino and Antonio Molina sent me go a long way toward clinching these objects as partially digested starch grains - maybe we can ask them to post a few of these (or I can, if they give me permission) for comparison. It looks like mine are simply further along in digestion. It's amazing to me that the outside is more resistant than the inside, but this would explain their hollowness.
Thanks again for all your help. If any of you are ever in Sackville, I owe you a genuine Canadian beer plus a pile of whatever seafood happens to be in season.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have an LKB 2088 that I am trying to resurrect. Are there any users out there who might be able to provide a copy of the manual, or be willing to answer a few specific questions?
The microtome is functioning, and we are trying to cut sections with a glass knives made on 1-inch glass with the LKB glass knife maker. The chuck that holds the knife is oversized for the glass, and the only way I can get the knife edge up to the section plane is by bringing the base (of the knife) up about a quarter inch (3-4mm) above the base of the holder. Is there a separate holder that I am missing, or can somebody tell me how to adjust the sample down so it meets the knife edge. I can cut sections with it, but with the base "floating" above the holder we are seeing a lot of chatter.
Thanks in advance.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 6, 23 -- From hagglundk1-at-nku.edu Fri Feb 24 13:45:04 2006 6, 23 -- Received: from mailfe2.nku.edu (exchange.nku.edu [192.122.237.68]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1OJj4Dt009917 6, 23 -- for {microscopy-at-microscopy.com} ; Fri, 24 Feb 2006 13:45:04 -0600 6, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 23 -- Fri, 24 Feb 2006 14:45:02 -0500 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: LKB 2088 Ultramicrotome question 6, 23 -- Date: Fri, 24 Feb 2006 14:45:01 -0500 6, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F35867D-at-mailfac1.hh.nku.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: LKB 2088 Ultramicrotome question 6, 23 -- Thread-Index: AcY5es05gg6goD1bQIufV5Adng0rAw== 6, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 24 Feb 2006 19:45:02.0424 (UTC) FILETIME=[CD90A580:01C6397A] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k1OJj4Dt009917 ==============================End of - Headers==============================
Is this the LKB I or LKB III? We have manuals for both.
Regarding the positioning of the knife in the ultramicrotome, it is CORRECT that the knife is raised above the base by about 3/4 inch. Reason: so that the knife holder can accommodate knives made using larger pieces of glass. To properly position the knife, flip up the little bar on the left side of the knife holder. This shows you the height where the knife edge should be placed. Tighten the knife while it is at this position. It should be snug but not excessively tight. If you are seeing chatter, it most likely is not due to a loose knife but other causes (vibration in the room, clearance angle of the knife, unsharp knife, loose components in the ultramicrotome, etc.)
JB
} I have an LKB 2088 that I am trying to resurrect. Are there any users } out there who might be able to provide a copy of the manual, or be } willing to answer a few specific questions? } } The microtome is functioning, and we are trying to cut sections with a } glass knives made on 1-inch glass with the LKB glass knife maker. The } chuck that holds the knife is oversized for the glass, and the only way } I can get the knife edge up to the section plane is by bringing the base } (of the knife) up about a quarter inch (3-4mm) above the base of the } holder. Is there a separate holder that I am missing, or can somebody } tell me how to adjust the sample down so it meets the knife edge. I can } cut sections with it, but with the base "floating" above the holder we } are seeing a lot of chatter.
-- ############################################################## 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============================== 6, 18 -- From bozzola-at-siu.edu Fri Feb 24 14:14:17 2006 6, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1OKEHQA029087 6, 18 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 24 Feb 2006 14:14:17 -0600 6, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 6, 18 -- by abbmta2.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k1OKEF60013623 6, 18 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 24 Feb 2006 14:14:17 -0600 (CST) 6, 18 -- Mime-Version: 1.0 6, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 6, 18 -- Message-Id: {p0611040bc025161c89c7-at-[131.230.177.142]} 6, 18 -- In-Reply-To: {200602241946.k1OJkBM8011796-at-ns.microscopy.com} 6, 18 -- References: {200602241946.k1OJkBM8011796-at-ns.microscopy.com} 6, 18 -- Date: Fri, 24 Feb 2006 14:14:12 -0600 6, 18 -- To: Microscopy-at-msa.microscopy.com 6, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 6, 18 -- Subject: Re: [Microscopy] LKB 2088 Ultramicrotome question 6, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
If people want to receive responses to their postings, could they please:
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2. Stop using anti-spam systems which require posters to to confirm responses. If you employer insists, I suggest you transfer your subscription to your personal, home PC.
This list is extremely well run and monitored - it doesn't need 'additional' security.
-- Larry Stoter
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==============================Original Headers============================== 6, 14 -- From larry-at-cymru.freewire.co.uk Fri Feb 24 15:06:58 2006 6, 14 -- Received: from get.freewire.net (get.freewire.net [195.184.229.250]) 6, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1OL6vkR006605 6, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 24 Feb 2006 15:06:57 -0600 6, 14 -- Received: from [217.154.252.45] (th6dc-217-154-252-45.dial.mistral.co.uk [217.154.252.45] (may be forged)) 6, 14 -- by get.freewire.net (8.11.6/8.11.6) with ESMTP id k1OL6te13389 6, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 24 Feb 2006 21:06:55 GMT 6, 14 -- Mime-Version: 1.0 6, 14 -- Message-Id: {p06210200c025236413fd-at-[217.154.255.187]} 6, 14 -- Date: Fri, 24 Feb 2006 21:07:35 +0000 6, 14 -- To: Microscopy-at-MSA.Microscopy.Com 6, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 6, 14 -- Subject: Out-of-Office and Anti-SPAM Measures 6, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
It has been interesting reading the replies you have received to your message. The general consensus is that everyone gets wrinkles in sections of Epon-embedded specimens and the discussion then centering on what method is best for removing them.
What no-one has mentioned is that if the specimens have been well-embedded in the resin (long infiltration), you will not have to worry about wrinkles in the sections. Therefore, you will have no worries about using ethylene dichloride or chloroform.
The other point that everyone missed is that you are performing immunolabeling on Epon-embedded sections. I know there are some published results using this resin (see works by Otterson from Oslo), but in general, there has been little success reported.
The most successful exponents of immunolabeling on Epon sections usually get their results from etching the resin from the section using chemicals that are much more harmful to your antigens than ethylene dichloride would be.
Why not try one of the other resins that have been designed for immunolabeling before attempting to perform it on Epon? I know that Lowicryl and LR White do not give the sort of text-book contrast you see using Epon (and presumably glutaraldehyde and osmium tetroxide), but they will give you an idea of how the antibody will work. You could then move onto the Epon sections to compare the results.
Best regards,
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
} From: {dsoren-at-umich.edu} } Reply-To: {dsoren-at-umich.edu} } Date: Thu, 23 Feb 2006 16:03:58 -0600 } To: {pwebster-at-hei.org} } Subject: [Microscopy] Wrinkles and ethylene dichloride } Does anyone know whether or not waving a magic wand, laden with ethylene dichloride, over wrinkled sections will destroy the tissue's antigenicity for post-embedding immunogold labeling? I'd love to relax out the wrinkles, but I have been hesitating to use the ethylene dichloride trick for fear of altering the tissue's antigenicity. The tissue is embedded in Epon.
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology
==============================Original Headers============================== 14, 19 -- From PWebster-at-hei.org Fri Feb 24 15:16:44 2006 14, 19 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 14, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1OLGg02016272 14, 19 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Feb 2006 15:16:44 -0600 14, 19 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 14, 19 -- Fri, 24 Feb 2006 21:13:35 +0000 14, 19 -- User-Agent: Microsoft-Entourage/11.2.1.051004 14, 19 -- Date: Fri, 24 Feb 2006 13:16:37 -0800 14, 19 -- Subject: Re: [Microscopy] Wrinkles and ethylene dichloride 14, 19 -- From: "Webster, Paul" {PWebster-at-hei.org} 14, 19 -- To: {dsoren-at-umich.edu} , {microscopy-at-msa.microscopy.com} 14, 19 -- Message-ID: {C024B7B5.855C%PWebster-at-hei.org} 14, 19 -- Thread-Topic: [Microscopy] Wrinkles and ethylene dichloride 14, 19 -- Thread-Index: AcY5h5iW12IvhKV6EdqtKwANk7Zh7g== 14, 19 -- In-Reply-To: {200602232203.k1NM3w1r020089-at-ns.microscopy.com} 14, 19 -- Mime-version: 1.0 14, 19 -- Content-type: text/plain; 14, 19 -- charset="US-ASCII" 14, 19 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
it says: HIGHLIGHTS / MILESTONES: 1978 Elektronenmikroskop EM 109 mit Trans-Faseroptik-Fotografiereinrichtung
in English: http://www.zeiss.com/4125681C00466C26/Contents-Frame/09FEFE9252D1059785256B75005EDDEC (Timetable, click on link } 1970-1979 {)
Regards,
Wolfgang Muss, Salzburg OR Dr. Wolfgang Muss EM-Lab =} 25 years in operation with a ZEISS EM 109 by 2nd of Feb.2006 {= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at --------------------------------------------------------------------------------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------- Forthcoming Meetings: 33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl
34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic
35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
---------- Von: wood-at-pw.usda.gov[SMTP:wood-at-pw.usda.gov] Antwort an: wood-at-pw.usda.gov Gesendet: Samstag, 25. Februar 2006 02:27 An: W.Muss-at-salk.at Betreff: [Microscopy] Zeiss EM 109 age?
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How old is the Zeiss EM 109??
Thanks
==============================Original Headers============================== 6, 19 -- From wood-at-pw.usda.gov Fri Feb 24 19:21:57 2006 6, 19 -- Received: from aggie.pw.usda.gov (pw.usda.gov [147.49.50.52]) 6, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1P1Luvg029485 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 24 Feb 2006 19:21:56 -0600 6, 19 -- Received: from (147.49.50.52) by DA32USCAAL1_AVS01.usda.gov via smtp 6, 19 -- id 13c8_e7fc81d2_a572_11da_8235_0014221728ad; 6, 19 -- Fri, 24 Feb 2006 20:19:50 +0000 6, 19 -- Received: from PW25-38ALXPDW.pw.usda.gov (pw25-38alxpdw.pw.usda.gov [147.49.4.121]) 6, 19 -- by aggie.pw.usda.gov (8.13.0/8.13.0) with ESMTP id k1P1LrRv020924 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 24 Feb 2006 17:21:55 -0800 (PST) 6, 19 -- Message-Id: {7.0.0.16.0.20060224172044.01ad4e10-at-pw.usda.gov} 6, 19 -- Message-Id: {7.0.0.16.0.20060224171943.01a49c28-at-pw.usda.gov} 6, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.0.16 6, 19 -- Date: Fri, 24 Feb 2006 17:21:51 -0800 6, 19 -- To: Microscopy-at-MSA.Microscopy.Com 6, 19 -- From: "Delilah F. Wood" {wood-at-pw.usda.gov} 6, 19 -- Subject: Zeiss EM 109 age? 6, 19 -- Mime-Version: 1.0 6, 19 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 24, 28 -- From W.Muss-at-salk.at Sat Feb 25 05:21:09 2006 24, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 24, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1PBL9NF022596 24, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sat, 25 Feb 2006 05:21:09 -0600 24, 28 -- Received: from localhost (localhost [127.0.0.1]) 24, 28 -- by hermes.lks.at (Postfix) with ESMTP id DF1155A9046; 24, 28 -- Sat, 25 Feb 2006 12:21:07 +0100 (CET) 24, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 24, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 24, 28 -- with ESMTP id 10123-02; Sat, 25 Feb 2006 12:21:07 +0100 (CET) 24, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 24, 28 -- by hermes.lks.at (Postfix) with SMTP id 841525A9044; 24, 28 -- Sat, 25 Feb 2006 12:21:07 +0100 (CET) 24, 28 -- Received: by localhost with Microsoft MAPI; Sat, 25 Feb 2006 12:21:06 +0100 24, 28 -- Message-ID: {01C63A05.F3E8B700.W.Muss-at-salk.at} 24, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 24, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 24, 28 -- To: "'wood-at-pw.usda.gov'" {wood-at-pw.usda.gov} 24, 28 -- Cc: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-MSA.Microscopy.Com} 24, 28 -- Subject: [Microscopy] RE: Zeiss EM 109 age? 24, 28 -- Date: Sat, 25 Feb 2006 12:21:04 +0100 24, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 24, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 24, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 24, 28 -- MIME-Version: 1.0 24, 28 -- Content-Type: text/plain; charset="us-ascii" 24, 28 -- Content-Transfer-Encoding: 7bit 24, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
I have a contrarian view. I think that these might be plant pollen. Check the plants where the animals would likely feed and see if you can find a match.
The holes are very characteristic of pollen as well as diatoms. If the animals are in a general environment, I would look at the plants.
I can put up some SEM pix of pollen that exhibit the holes theme.
gary g.
At 09:14 AM 2/24/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Sat Feb 25 19:14:54 2006 11, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1Q1EsWm012382 11, 20 -- for {microscopy-at-microscopy.com} ; Sat, 25 Feb 2006 19:14:54 -0600 11, 20 -- Received: (qmail 25982 invoked from network); 25 Feb 2006 17:14:53 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 25978, pid: 25980, t: 0.1634s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp1 with SMTP; 25 Feb 2006 17:14:53 -0800 11, 20 -- Message-Id: {6.2.3.4.2.20060225171131.020361a8-at-mail.calweb.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 11, 20 -- Date: Sat, 25 Feb 2006 17:14:56 -0800 11, 20 -- To: jehrman-at-mta.ca 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] mystery object - thanks for all the help 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200602241714.k1OHE1bh001928-at-ns.microscopy.com} 11, 20 -- References: {200602241714.k1OHE1bh001928-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I have a contrarian view. I think that these might be plant pollen. Check the plants where the animals would likely feed and see if you can find a match.
The holes are very characteristic of pollen as well as diatoms. If the animals are in a general environment, I would look at the plants.
I can put up some SEM pix of pollen that exhibit the holes theme.
gary g.
At 09:14 AM 2/24/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 11, 18 -- From gary-at-gaugler.com Sat Feb 25 20:51:02 2006 11, 18 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1Q2p2hE022779 11, 18 -- for {microscopy-at-microscopy.com} ; Sat, 25 Feb 2006 20:51:02 -0600 11, 18 -- Received: (qmail 32722 invoked from network); 25 Feb 2006 18:51:00 -0800 11, 18 -- Received: by simscan 1.1.0 ppid: 32719, pid: 32720, t: 0.1659s 11, 18 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 18 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 18 -- by qsmtp2 with SMTP; 25 Feb 2006 18:51:00 -0800 11, 18 -- Message-Id: {6.2.3.4.2.20060225185059.02034680-at-mail.calweb.com} 11, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 11, 18 -- Date: Sat, 25 Feb 2006 18:51:04 -0800 11, 18 -- To: jehrman-at-mta.ca 11, 18 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 18 -- Subject: Re: [Microscopy] mystery object - thanks for all the help 11, 18 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 18 -- Mime-Version: 1.0 11, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I have been asked to estimate what SEM and TEM labs would cost. As I am alight microscopist, I do not know how to specify these labs. I would like to ask listmembers who have assembled these labs to give me a rough idea of what they will cost.
I realize that this question is too broad as it is posed. There are too many variables. Therefore, I would appreciate it if people who run a central facility for electron microscopy would be willing to tell me what they have and about what the major items cost. To the nearest $50,000 is fine.
I have done some searching, so I have found descriptions of some facilities, but I have not found cost information on these sites.
I some kind souls would also tell me what they recommend be included in such labs, I would also be most appreciative.
I apologize in advance if this inquiry is a bit off the wall, and thanks for any assistance you can provide.
--aryeh -- Aryeh Weiss School of Engineering Bar Ilan University Ramat Gan 52900 Israel
Ph: 972-3-5317638 FAX: 972-3-5340697
==============================Original Headers============================== 8, 29 -- From aryeh-at-cc.huji.ac.il Sun Feb 26 06:57:13 2006 8, 29 -- Received: from tamar.os.biu.ac.il (tamar.os.biu.ac.il [132.70.60.24]) 8, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1QCvCK2015827 8, 29 -- for {Microscopy-at-microscopy.com} ; Sun, 26 Feb 2006 06:57:13 -0600 8, 29 -- Received: from ismss-2.biu.ac.il (ismss-2.biu.ac.il [132.70.84.151]) 8, 29 -- by tamar.os.biu.ac.il (8.12.11/8.12.11/BIU) with ESMTP id k1QCv6p06443176 8, 29 -- for {Microscopy-at-microscopy.com} ; Sun, 26 Feb 2006 14:57:11 +0200 8, 29 -- Received: from cc.huji.ac.il ([132.70.133.31]RDNS failed) by 8, 29 -- ismss-2.biu.ac.il with InterScan Message Security Suite; Sun, 26 Feb 2006 8, 29 -- 14:57:24 +0200 8, 29 -- Message-ID: {4401A59C.4070306-at-cc.huji.ac.il} 8, 29 -- Date: Sun, 26 Feb 2006 14:57:00 +0200 8, 29 -- From: Aryeh Weiss {aryeh-at-cc.huji.ac.il} 8, 29 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) 8, 29 -- Gecko/20030624 Netscape/7.1 (ax) 8, 29 -- X-Accept-Language: en-us, en, he 8, 29 -- MIME-Version: 1.0 8, 29 -- To: Microscopy-at-microscopy.com 8, 29 -- Subject: cost of TEM, SEM and AFM 8, 29 -- Content-Type: text/plain; 8, 29 -- charset=us-ascii; 8, 29 -- format=flowed 8, 29 -- Content-Transfer-Encoding: 7bit 8, 29 -- X-imss-version: 2.038 8, 29 -- X-imss-result: Passed 8, 29 -- X-imss-scanInfo: M:P L:E SM:0 8, 29 -- X-imss-tmaseResult: TT:0 TS:0.0000 TC:00 TRN:0 TV:3.52.1006(14288.000) 8, 29 -- X-imss-scores: Clean:13.21832 C:2 M:3 S:5 R:5 8, 29 -- X-imss-settings: Baseline:2 C:1 M:1 S:1 R:1 (0.1500 0.1500) ==============================End of - Headers==============================
When calculate your budget don't forget to include the cost of services (air, water, air-conditioning etc.), people, consumables, and service contracts.
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 07:01 AM 2/26/2006, aryeh-at-cc.huji.ac.il wrote:
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==============================Original Headers============================== 12, 17 -- From bfoster-at-mme1.com Mon Feb 27 10:06:09 2006 12, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 12, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1RG68U2019821 12, 17 -- for {microscopy-at-microscopy.com} ; Mon, 27 Feb 2006 10:06:09 -0600 12, 17 -- Received: (qmail 30793 invoked from network); 27 Feb 2006 11:07:54 -0500 12, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 12, 17 -- by enterprise.5starpro.com with SMTP; 27 Feb 2006 11:07:54 -0500 12, 17 -- Message-Id: {7.0.1.0.0.20060227100451.02090b30-at-mme1.com} 12, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 17 -- Date: Mon, 27 Feb 2006 10:06:14 -0600 12, 17 -- To: aryeh-at-cc.huji.ac.il, Microscopy Listserver {microscopy-at-microscopy.com} 12, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 12, 17 -- Subject: Re: [Microscopy] cost of TEM, SEM and AFM 12, 17 -- In-Reply-To: {200602261301.k1QD1KPN020084-at-ns.microscopy.com} 12, 17 -- References: {200602261301.k1QD1KPN020084-at-ns.microscopy.com} 12, 17 -- Mime-Version: 1.0 12, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
} Including everything Barbara suggests (particularly service } contracts), and starting from scratch with new instrumentation, } you'll need to budget at least US$1M. With used equipment it will } be somewhat less expensive, probably by about 50%
Good luck!
Peter
} ---------------------------------------------------------------------------- } } Hi, Aryeh } } When calculate your budget don't forget to include the cost of } services (air, water, air-conditioning etc.), people, consumables, } and service contracts. } } 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 07:01 AM 2/26/2006, aryeh-at-cc.huji.ac.il wrote: } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 16 -- From p.ingram-at-voice.cellbio.duke.edu Mon Feb 27 11:38:10 2006 7, 16 -- Received: from eddings.acpub.duke.edu (eddings.acpub.duke.edu [152.3.233.76]) 7, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1RHcAbH030669 7, 16 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Feb 2006 11:38:10 -0600 7, 16 -- Received: from [152.3.54.111] (dhcp111.fel.duke.edu [152.3.54.111]) 7, 16 -- by eddings.acpub.duke.edu (8.12.10/8.12.10/Duke-5.0.0) with ESMTP id k1RHc9gX005084; 7, 16 -- Mon, 27 Feb 2006 12:38:09 -0500 (EST) 7, 16 -- Mime-Version: 1.0 7, 16 -- Message-Id: {a06002001c028e81f14cb-at-[152.3.54.111]} 7, 16 -- In-Reply-To: {200602271611.k1RGBjc8025688-at-ns.microscopy.com} 7, 16 -- References: {200602271611.k1RGBjc8025688-at-ns.microscopy.com} 7, 16 -- Date: Mon, 27 Feb 2006 12:37:57 -0500 7, 16 -- To: Microscopy-at-microscopy.com 7, 16 -- From: Peter Ingram {p.ingram-at-voice.cellbio.duke.edu} 7, 16 -- Subject: [Microscopy] Re: cost of TEM, SEM and AFM 7, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Allow to ask a question perhaps not usual for this forum:
We do have a patient suffering from several pain due to a post-vaccination problem (due to accumulation of Al-precipitates in intermuscular macrophages) , namely
macrophagic myofasciitis,
luckily NOT systemic or generalized.
If there is anybody out there who has knowledge about a specific and reliable treatment for such a condition I greatly should appreciate your comment. (Unfortunately I was not able to localize any reference for } successful { treatment).
If -perhaps- I have broken rules for using this listserver, please apologize. It is just to seek help for a young patient.
Thank you and cordially yours
Wolfgang Muss, PhD. Salzburg, Austria
==============================Original Headers============================== 10, 28 -- From W.Muss-at-salk.at Mon Feb 27 12:07:37 2006 10, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 10, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1RI7YQ1008343 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Feb 2006 12:07:35 -0600 10, 28 -- Received: from localhost (localhost [127.0.0.1]) 10, 28 -- by hermes.lks.at (Postfix) with ESMTP id 7AC035A9046 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Feb 2006 19:07:31 +0100 (CET) 10, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 10, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 10, 28 -- with ESMTP id 59654-01 for {Microscopy-at-MSA.Microscopy.Com} ; 10, 28 -- Mon, 27 Feb 2006 19:07:31 +0100 (CET) 10, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 10, 28 -- by hermes.lks.at (Postfix) with SMTP id EA7395A9020 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Feb 2006 19:07:30 +0100 (CET) 10, 28 -- Received: by localhost with Microsoft MAPI; Mon, 27 Feb 2006 19:07:29 +0100 10, 28 -- Message-ID: {01C63BD1.0DFB1440.W.Muss-at-salk.at} 10, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 10, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 10, 28 -- To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-MSA.Microscopy.Com} 10, 28 -- Subject: Vaccination complications: macrophagic myofasciitis 10, 28 -- Date: Mon, 27 Feb 2006 19:07:28 +0100 10, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 10, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 10, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 10, 28 -- MIME-Version: 1.0 10, 28 -- Content-Type: text/plain; charset="us-ascii" 10, 28 -- Content-Transfer-Encoding: 7bit 10, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
A critical consideration that is often times forgotten is the cost of specimen preparation equipment. As a manufacturer of specimen prep equipment, I obviously have a slight bias. However, the cost for a state of the art specimen preparation facility is not trivial and should certainly be considered up front. I would welcome the opportunity to discuss this with you off-line and can give you a pretty accurate estimate after discussing with you the sample types and volume of samples you anticipate.
Best regards-
David
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bfoster-at-mme1.com wrote:
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==============================Original Headers============================== 17, 21 -- From henriks-at-southbaytech.com Mon Feb 27 13:08:34 2006 17, 21 -- Received: from ylpvm25.prodigy.net (ylpvm25-ext.prodigy.net [207.115.57.56]) 17, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1RJ8YRL018800 17, 21 -- for {microscopy-at-msa.microscopy.com} ; Mon, 27 Feb 2006 13:08:34 -0600 17, 21 -- Received: from southbaytech.com (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 17, 21 -- (authenticated bits=0) 17, 21 -- by ylpvm25.prodigy.net (smtpauth/dk/flock 8.13.4/8.13.4) with ESMTP id k1RJ78E2005501; 17, 21 -- Mon, 27 Feb 2006 14:07:11 -0500 17, 21 -- Message-ID: {44034E3E.8050000-at-southbaytech.com} 17, 21 -- Date: Mon, 27 Feb 2006 11:08:46 -0800 17, 21 -- From: David Henriks {henriks-at-southbaytech.com} 17, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 17, 21 -- X-Accept-Language: en-us, en 17, 21 -- MIME-Version: 1.0 17, 21 -- To: Microscopy Listerver {microscopy-at-msa.microscopy.com} 17, 21 -- CC: aryeh-at-cc.huji.ac.il 17, 21 -- Subject: Re: [Microscopy] Re: cost of TEM, SEM and AFM 17, 21 -- References: {200602271609.k1RG9IFr022784-at-ns.microscopy.com} 17, 21 -- In-Reply-To: {200602271609.k1RG9IFr022784-at-ns.microscopy.com} 17, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 17, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hello, Our lab is still using conventional photography. Recently I was informed that Kodak Ektamatic Stabilizer S30 was discontinued. Do you Lisetservers now if there is a substitute for it, is there any photographic company out there that still supplies photographic reagents? Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Mon Feb 27 13:39:12 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1RJdCp5028712 1, 22 -- for {microscopy-at-microscopy.com} ; Mon, 27 Feb 2006 13:39:12 -0600 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1FDoD9-00015V-02 1, 22 -- for {microscopy-at-microscopy.com} ; Mon, 27 Feb 2006 15:39:11 -0400 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 27 Feb 06 15:39:11 -0400 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 27 Feb 06 15:38:08 -0400 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Mon, 27 Feb 2006 15:33:51 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: Dark room reagents 1, 22 -- Message-ID: {44031BDF.3093.2F271B-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both johnsonk-at-uhd.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: We have an old Nikon Labophot-Pol microscope. Lately, the stage will sink under its own weight, and will not respond to coarse adjustments, regardless of the torque adjustment ring setting. I want to ask the listserv if there were an easy way to fix this, or if it requires opening the housing. Any help or advice would be appreciated.
In case you are interested in development of high-content microscopy imaging and if you would be interested to join an international research institute in Dresden ( people from more then 30 countries), please have a look at the following advert : http://www.mpi-cbg.de/research/jobs/tds.html
At the HT-Technology Development Studio (TDS) of the MPI-CBG in Dresden we are developing phenotypic assays and implementing high-content screens in cultured cells and model organisms with the aid of robotic systems and automated microscopy to discover novel gene functions by RNA interference and small molecule compounds in close collaboration with internal and external scientific groups. The facility operates in partnership with Cellomics Inc., Evotec Technologies GmbH, and Definiens AG to develop automated microscopy and software for pattern recognition and image analysis. The TDS was established by Drs Marino Zerial and Ivan Baines, and is headed up by Dr. Eberhard Krausz (tds.mpi-cbg.de/webtds/; Pelkmans et al., Nature 2005, 436:78-86).
We are currently seeking to fill at least one postdoctoral scientist position responsible for
Cell-based Assay Development siRNA Transfection High-Content Screening
You are a skilled cell biologist with some years of experience in fluorescent microscopy and cell-based assays. A strong scientific background in cell biology is essential. Some experience in project management and supervision of technicians would be a plus. Excellent communication skills, goal-orientated working style and the ability to work as part of a multidisciplinary team are required. Our working language is English.
Contracts are limited initially to 2 years. Compensation follows the applicable German tariff for employees in civil service according to qualification, experience and job description.
Please send your application to
Max Planck Institute of Molecular Cell Biology and Genetics Personnel Department Pfotenhauerstr. 108 D - 01307 Dresden Germany
E-mail: aotto-at-mpi-cbg.de
___________________________________________________________ To help you stay safe and secure online, we've developed the all new Yahoo! Security Centre. http://uk.security.yahoo.com
==============================Original Headers============================== 13, 18 -- From jb_sanderson-at-yahoo.com Tue Feb 28 04:45:03 2006 13, 18 -- Received: from web42006.mail.yahoo.com (web42006.mail.yahoo.com [66.218.93.174]) 13, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1SAj3nP006735 13, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 28 Feb 2006 04:45:03 -0600 13, 18 -- Received: (qmail 61734 invoked by uid 60001); 28 Feb 2006 10:45:02 -0000 13, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 13, 18 -- s=s1024; d=yahoo.com; 13, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 13, 18 -- b=QLzFUUEWzjlByYeaT+loOItgPP2lNEb7nqA2PDpIa7WYHdV7lzQOiDo87VfGVwIO7XaDFBTr5JAAsvdPidTF0xl5/b4SrZDtR44GLbj/ywhBY510PpunkJ8aE9CkygHFLHRsOCwn5oRLpgpkUoGTHfvyf2sn4h8SMUzqAS17o/g= ; 13, 18 -- Message-ID: {20060228104502.61729.qmail-at-web42006.mail.yahoo.com} 13, 18 -- Received: from [84.13.68.115] by web42006.mail.yahoo.com via HTTP; Tue, 28 Feb 2006 10:45:01 GMT 13, 18 -- Date: Tue, 28 Feb 2006 10:45:01 +0000 (GMT) 13, 18 -- From: Jeremy Sanderson {jb_sanderson-at-yahoo.com} 13, 18 -- Subject: High-content Microscopy Screening Position in Dresden 13, 18 -- To: CONFOCAL-at-LISTSERV.BUFFALO.EDU, Microscopy-at-MSA.Microscopy.Com 13, 18 -- MIME-Version: 1.0 13, 18 -- Content-Type: text/plain; charset=iso-8859-1 13, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Here is the March 2006 Microscopy Today table of contents. I will close the subscription list for this issue on Thursday March 7, 2006.
Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$50 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Ron Anderson, Editor =======================================================
Microscopes Reveal Prehistoric High Technology Stephen W. Carmichael, Mayo Clinic
The Influence of Microspectroscopy on Evaluating and Analyzing Forensic Evidence John A. Reffner and Pauline E. Leary, Smiths Detection, Danbury, CT
The Value of S/TEM: Matching Solutions, Applications, and Economics Dominique Hubert, FEI Company, Eindhoven, The Netherlands
Brief Review: Basic Properties and Applications of Carbon Nanotubes Supapan Seraphin, The University of Arizona, Tucson, Arizona
Assessing Microalgal Morphology from Century-Old Herbarium Sheets using SEM Linton, E. W.1, Farmer, M. A.2, & Triemer, R. E.,1 1) Michigan State University, East Lansing, MI & 2) The University of Georgia, Athens, GA
Microscopy in Ecuador Alwyn Eades, Lehigh University, Bethlehem, PA
Overcoming Severe XEDS Peak Overlaps with the AEM Scott D. Walck, South Bay Technology, Inc., San Clemente, CA
A Comparison of Photomicrographs Imaged Through a Late 18th C. Thomas Ribright, Cuff-Type, Brass Microscope and a Modern Olympus Optical Microscope By D.Jones* and J.Reid.**, *Retired Research Microbiologist Aberdeen, **School of Physics, The University, Aberdeen, Scotland
Computationally Mediated Experimental Science Nestor J. Zaluzec, Argonne National Laboratory, Argonne, Il
The Day Atomic Resolution Microscopy Happened Allan J. Melmed, Custom Probes Unlimited, Terra Alta, WV
Rhodamine Fluorescence After 15-year Storage in Methyl Salicylate Philip L. Hertzler, Dept. of Biology, Central Michigan University
When Point To Point Is Not Enough Carol Heckman, Marilyn Cayer, Mita Varghese, Bowling Green State University, Bowling Green, OH
Making Replicas of Surfaces for TEM and SEM Mary Mager, University of British Columbia
Industry News
NetNotes Topics:
LM: Refractive index of organelles
IMMUNOCYTOCHEMISTRY - Permount and FITC
IMMUNOCYTOCHEMISTRY - Bacterial permeabilization
TEM SAMPLE PREPARATION – Embedding tissue culture cells
TEM SAMPLE PREPARATION – making PVA and PVAH adhesives
MICROTOMY – Cleaning copper grids
MICROTOMY – Glue for serial sectioning
MICROTOMY – Making Formvar films
TEM SAMPLE PREPARATION – Negative staining pili
SEM SAMPLE PREPARATION – Critical point drying
SEM SAMPLE PREPARATION - Magnetic etching
SEM – Image distortion
TEM: LR White contrast
TEM - Colloidal Silica Particle Size Distribution)
TEM- Beam problem
TEM - Starting ion pumps without cooling water
EDS - Ge EDS detector
EDS - Oil on detector window
AFM - Sample preparation
Index of Advertisers
==============================Original Headers============================== 42, 18 -- From microscopytoday-at-tampabay.rr.com Tue Feb 28 11:14:43 2006 42, 18 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01-smtplb.tampabay.rr.com [65.32.5.131]) 42, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1SHEhMC030864 42, 18 -- for {Microscopy-at-Microscopy.Com} ; Tue, 28 Feb 2006 11:14:43 -0600 42, 18 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 42, 18 -- by ms-smtp-01.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k1SHEcRX008925; 42, 18 -- Tue, 28 Feb 2006 12:14:41 -0500 (EST) 42, 18 -- Message-ID: {440484FB.8070201-at-tampabay.rr.com} 42, 18 -- Date: Tue, 28 Feb 2006 12:14:35 -0500 42, 18 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 42, 18 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 42, 18 -- MIME-Version: 1.0 42, 18 -- To: Listserver {Microscopy-at-Microscopy.Com} , 42, 18 -- Confocal Listserver {confocal-at-listserv.buffalo.edu} 42, 18 -- Subject: Microscopy Today March 2006 Table of Contents 42, 18 -- Content-Type: text/plain; charset=windows-1252; format=flowed 42, 18 -- Content-Transfer-Encoding: 8bit 42, 18 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Southern California Society for Microscopy and Microanalysis 2006 Spring Meeting
Call for Posters
Date:May 4, 2006 Place: Golleher Alumni House California State University at Fullerton
The Southern California Society for Microscopy and Microanalysis extends an invitation for your attendance at the 2006 Spring meeting at the California State University at Fullerton.
This is an evening meeting which will feature invited speakers, experts from different fields (physical and biological sciences as well as instrumentation) who will present their research on the application of microscopy. Exhibits from several commercial vendors in the field of microscopy will also be held in conjunction with this meeting.
We would like to invite students to participate in our Student Poster Competition and share their research results with our society members and guests. Any research (biological or physical) with the application of any type of microscopy is welcome.
An abstract of the poster should be submitted in electronic format. There is no particular format, although the MSA format is recommended and up to two pages of text is acceptable.
Awards will be given to the top three student posters. Posters with a registered student as the first author qualify for the competition.
A complementary dinner will be provided to all registered participants; light refreshments and wine will also be served during the social hour and during the poster session.
Please submit your abstract in PDF format to: Krassimir Bozhilov, bozhilov-at-ucr.edu, (951)-827 2998. Please contact us or check our web site at www.scsmm.org should you need further assistance.
Deadline for Submission: April 15, 2006
==============================Original Headers============================== 12, 19 -- From bozhilov-at-ucr.edu Tue Feb 28 11:45:19 2006 12, 19 -- Received: from sentry.ucr.edu (sentry.ucr.edu [138.23.226.224]) 12, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1SHjJQs008557 12, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 11:45:19 -0600 12, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 12, 19 -- by sentry.ucr.edu (MOS 3.5.9-GR) 12, 19 -- with ESMTP id DPP64426 (AUTH via LOGINBEFORESMTP) 12, 19 -- for {microscopy-at-microscopy.com} ; 12, 19 -- Tue, 28 Feb 2006 09:45:14 -0800 (PST) 12, 19 -- Mime-Version: 1.0 (Apple Message framework v746.2) 12, 19 -- Content-Transfer-Encoding: 7bit 12, 19 -- Message-Id: {0B83D955-2E5D-4B83-882C-6BB66CF8E05F-at-ucr.edu} 12, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 19 -- To: microscopy-at-microscopy.com 12, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 12, 19 -- Subject: SCSMM 2006 Spring Meeting 12, 19 -- Date: Tue, 28 Feb 2006 09:45:13 -0800 12, 19 -- X-Mailer: Apple Mail (2.746.2) 12, 19 -- X-Junkmail-Status: score=0/65, host=sentry.ucr.edu ==============================End of - Headers==============================
Hello listservers, Does anyone have experience using fluorescent latex beads for epifluorescence micrography? My lab is trying to perform correlative electron and fluorescence studies using fluorescently-labeled latex beads (that are likewise visible at the electron microscope) but none of the students in our lab have experience with such probe.
Any suggestions on which company may have available latex beads that can be used as EM and fluorescent probes?
Thanks a lot! You all have been truly generous with extending technical assistance :)
-Carlo
==============================Original Headers============================== 5, 32 -- From cbalane-at-wesleyan.edu Tue Feb 28 14:51:22 2006 5, 32 -- Received: from post1.wesleyan.edu (post1.wesleyan.edu [129.133.6.131]) 5, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1SKpM5N021399 5, 32 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 14:51:22 -0600 5, 32 -- Received: from pony2.wesleyan.edu (pony2.wesleyan.edu [129.133.6.193]) 5, 32 -- by post1.wesleyan.edu (8.12.11/8.12.11) with ESMTP id k1SKpXTs009611 5, 32 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 15:51:33 -0500 5, 32 -- Received: from pony2.wesleyan.edu (pony2.wesleyan.edu [127.0.0.1]) 5, 32 -- by pony2.wesleyan.edu (8.12.11/8.12.11) with ESMTP id k1SKmtpU008585 5, 32 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 15:48:55 -0500 5, 32 -- Received: (from apache-at-localhost) 5, 32 -- by pony2.wesleyan.edu (8.12.11/8.12.11/Submit) id k1SKmtMu008583; 5, 32 -- Tue, 28 Feb 2006 15:48:55 -0500 5, 32 -- Received: from 129.133.93.129 5, 32 -- (SquirrelMail authenticated user cbalane); 5, 32 -- by webmail.wesleyan.edu with HTTP; 5, 32 -- Tue, 28 Feb 2006 15:48:55 -0500 (EST) 5, 32 -- Message-ID: {4128.129.133.93.129.1141159735.squirrel-at-129.133.93.129} 5, 32 -- Date: Tue, 28 Feb 2006 15:48:55 -0500 (EST) 5, 32 -- Subject: latex beads for EM 5, 32 -- From: "Carlo Franco Balane-Bolivar" {cbalane-at-wesleyan.edu} 5, 32 -- To: microscopy-at-microscopy.com 5, 32 -- User-Agent: SquirrelMail/1.4.3a-0.e3.1 5, 32 -- X-Mailer: SquirrelMail/1.4.3a-0.e3.1 5, 32 -- MIME-Version: 1.0 5, 32 -- Content-Type: text/plain;charset=iso-8859-1 5, 32 -- Content-Transfer-Encoding: 8bit 5, 32 -- X-Priority: 3 (Normal) 5, 32 -- Importance: Normal 5, 32 -- X-Wesleyan-MailScanner-Information: Please contact the ISP for more information 5, 32 -- X-Wesleyan-MailScanner: Found to be clean 5, 32 -- X-MailScanner-From: cbalane-at-wesleyan.edu ==============================End of - Headers==============================
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Email: zzhang-at-uwyo.edu Name: Zhaojie Zhang
Organization: University of Wyoming
Title-Subject: [Filtered] 3rd party TEM repair
Question: Due to the budget constrain, I have to drop the service contract for my Hitachi H7000 TEM. I am now looking for a 3rd party TEM repair and service that serves NW region (Wyoming). Comercial venders are welcome to contact me offline.
Zhaojie Zhang, Ph.D. Director, Microscopy Core Facility University of Wyoming Laramie, WY, 82071 307-766-3038 zzhang-at-uwyo.edu
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Email: randy-nessler-at-uiowa.edu Name: Randy Nessler
Organization: University of Iowa
Title-Subject: [Filtered] Fluorine removal?
Question: I have been asked to post this question for a collegue. It appears that he might be getting fluorine contamination of his XPS samples when they are in a processing chamber. The chamber is lined with glass. Is there a suitable reagent to clean this chamber out with to remove any residual fluorine? "Baking" the chamber hasn't minimied the problem. Thanks, Randy
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Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] Re: Manganese
Question: [Commercial disclaimer - we make silver enhancement reagents]
Hello Hong:
A similar question came up on the Confocal Listserver a while ago, so I dug up my answer...
I suggest trying silver enhancement - transition metal ions are thought to be one cause of background deposition in the silver enhancement reaction when silver enhancing gold particles for blots and light microscopy; we sometimes wash with a chelating agent such as an EDTA (ethylene diamine tetraacetic acid) salt to remove or chelate them before silver enhancement.
For activity towards silver enhancement, what we look for is an active redox chemistry where it's likely that one of the redox couples of the target can couple with that of the silver/hydroquinone reduction. Manganese is an excellent candidate for this (as is iron) because of its extensive redox chemistry; compounds are known for every oxidation state from +2 to +7, so there's a good chance that it has some reactivity towards silver enhancement. It's certainly worth trying.
I couldn't find any references to silver enhancement of manganese (Gorm Danscher is a good name to search, since he has used silver enhancement - or variations of the method - to localize many other metals, and may have some observations about the reactivity or interference of manganese in these processes). There is, however, a reference to manganese dioxide absorbing silver in water, which suggests that it could react with silver enhancement:
Anderson, J. B.; Jenne, E. A., and Chao, T. T.: The sorption of silver by poorly crystallized manganese oxides. Geochimica et Cosmochimica Acta, 37, 611-622 (1973).
Speaking from experience making manganese compounds in a previous life...Manganese likes to form manganese dioxide, which is an insoluble black precipitate and would probably be good for nucleating silver deposition. You don't mention what form the manganese is in, but treatment of your specimens with an oxidizing agent could generate manganese dioxide.
Hope this helps,
Rick Powell
***************************************************************************************** Richard D. Powel rpowell-at-nanoprobes.com * www.nanoprobes.com
NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA *****************************************************************************************
____________________________________
Dear All:
We have a PI here wants to localize manganese in tissue. Does anyone know a way of doing this without using X-ray microanalysis? Thank you.
There is a product called FluoroNanogold from nanoprobes. It is good for these things. I have never had any luck with latex beads in the TEM. David
On Feb 28, 2006, at 1:55 PM, cbalane-at-wesleyan.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Hello listservers, } Does anyone have experience using fluorescent latex beads for } epifluorescence micrography? My lab is trying to perform correlative } electron and fluorescence studies using fluorescently-labeled latex } beads } (that are likewise visible at the electron microscope) but none of the } students in our lab have experience with such probe. } } Any suggestions on which company may have available latex beads } that can } be used as EM and fluorescent probes? } } Thanks a lot! You all have been truly generous with extending } technical } assistance :) } } -Carlo } } } ==============================Original } Headers============================== } 5, 32 -- From cbalane-at-wesleyan.edu Tue Feb 28 14:51:22 2006 } 5, 32 -- Received: from post1.wesleyan.edu (post1.wesleyan.edu } [129.133.6.131]) } 5, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k1SKpM5N021399 } 5, 32 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 } 14:51:22 -0600 } 5, 32 -- Received: from pony2.wesleyan.edu (pony2.wesleyan.edu } [129.133.6.193]) } 5, 32 -- by post1.wesleyan.edu (8.12.11/8.12.11) with ESMTP id } k1SKpXTs009611 } 5, 32 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 } 15:51:33 -0500 } 5, 32 -- Received: from pony2.wesleyan.edu (pony2.wesleyan.edu } [127.0.0.1]) } 5, 32 -- by pony2.wesleyan.edu (8.12.11/8.12.11) with ESMTP id } k1SKmtpU008585 } 5, 32 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 } 15:48:55 -0500 } 5, 32 -- Received: (from apache-at-localhost) } 5, 32 -- by pony2.wesleyan.edu (8.12.11/8.12.11/Submit) id } k1SKmtMu008583; } 5, 32 -- Tue, 28 Feb 2006 15:48:55 -0500 } 5, 32 -- Received: from 129.133.93.129 } 5, 32 -- (SquirrelMail authenticated user cbalane); } 5, 32 -- by webmail.wesleyan.edu with HTTP; } 5, 32 -- Tue, 28 Feb 2006 15:48:55 -0500 (EST) } 5, 32 -- Message-ID: } {4128.129.133.93.129.1141159735.squirrel-at-129.133.93.129} } 5, 32 -- Date: Tue, 28 Feb 2006 15:48:55 -0500 (EST) } 5, 32 -- Subject: latex beads for EM } 5, 32 -- From: "Carlo Franco Balane-Bolivar" {cbalane-at-wesleyan.edu} } 5, 32 -- To: microscopy-at-microscopy.com } 5, 32 -- User-Agent: SquirrelMail/1.4.3a-0.e3.1 } 5, 32 -- X-Mailer: SquirrelMail/1.4.3a-0.e3.1 } 5, 32 -- MIME-Version: 1.0 } 5, 32 -- Content-Type: text/plain;charset=iso-8859-1 } 5, 32 -- Content-Transfer-Encoding: 8bit } 5, 32 -- X-Priority: 3 (Normal) } 5, 32 -- Importance: Normal } 5, 32 -- X-Wesleyan-MailScanner-Information: Please contact the ISP } for more information } 5, 32 -- X-Wesleyan-MailScanner: Found to be clean } 5, 32 -- X-MailScanner-From: cbalane-at-wesleyan.edu } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 18 -- From Elliott-at-arizona.edu Tue Feb 28 19:09:06 2006 5, 18 -- Received: from elliott-server.doctorelliott.us (doctorelliott.us [70.57.226.104]) 5, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21196jj009192 5, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 28 Feb 2006 19:09:06 -0600 5, 18 -- Received: from [192.168.0.30] (unknown [192.168.0.30]) 5, 18 -- by elliott-server.doctorelliott.us (Postfix) with ESMTP id B85C91A41FD 5, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 28 Feb 2006 18:08:58 -0700 (MST) 5, 18 -- Mime-Version: 1.0 (Apple Message framework v746.2) 5, 18 -- In-Reply-To: {200602282055.k1SKtcnS027103-at-ns.microscopy.com} 5, 18 -- References: {200602282055.k1SKtcnS027103-at-ns.microscopy.com} 5, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 18 -- Message-Id: {B9C07F52-8323-472D-89A7-D7EF3233BDD4-at-arizona.edu} 5, 18 -- Content-Transfer-Encoding: 7bit 5, 18 -- From: David Elliott {Elliott-at-arizona.edu} 5, 18 -- Subject: Re: [Microscopy] latex beads for EM 5, 18 -- Date: Tue, 28 Feb 2006 18:08:56 -0700 5, 18 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 18 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
Zhaojie Zhang wrote: ====================================================================== Question: Due to the budget constrain, I have to drop the service contract for my Hitachi H7000 TEM. I am now looking for a 3rd party TEM repair and service that serves NW region (Wyoming). Comercial venders are welcome to contact me offline. ===================================================================== We maintain a reasonably up-to-date listing of third party services firms working on TEMs on URL http://www.2spi.com/catalog/hot-service6.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============================== 12, 30 -- From cgarber-at-2spi.com Tue Feb 28 21:28:54 2006 12, 30 -- Received: from s-utl01-atpop.stsn.net (s-utl01-atpop.stsn.net [72.254.128.201]) 12, 30 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k213SsOR020325 12, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Feb 2006 21:28:54 -0600 12, 30 -- Received: from s-utl01-atpop.stsn.net ([127.0.0.1]) 12, 30 -- by s-utl01-atpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006022822285307599 12, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Feb 2006 22:28:53 -0500 12, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 12, 30 -- tests=ALL_TRUSTED: -2.867,AWL: 0.059,BAYES_00: -1.665, 12, 30 -- SARE_RECV_ADDR: 0.027 12, 30 -- X-Spam-Level: 12, 30 -- Received: from ibm1x23g2abfyg ([10.10.23.158]) 12, 30 -- by s-utl01-atpop.stsn.net 12, 30 -- for microscopy-at-msa.microscopy.com; 12, 30 -- Tue, 28 Feb 2006 22:28:52 -0500 12, 30 -- Message-ID: {10de01c63ce0$40ab8040$9e170a0a-at-ibm1x23g2abfyg} 12, 30 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 12, 30 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 12, 30 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 30 -- Subject: Third party firms for TEM repairs and servicing 12, 30 -- Date: Tue, 28 Feb 2006 22:28:37 -0500 12, 30 -- MIME-Version: 1.0 12, 30 -- Content-Type: text/plain; 12, 30 -- charset="Windows-1252" 12, 30 -- X-Priority: 3 12, 30 -- X-MSMail-Priority: Normal 12, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 12, 30 -- X-Mimeole: Produced By Microsoft MimeOLE V6.00.2900.2180 12, 30 -- Content-Transfer-Encoding: 8bit 12, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k213SsOR020325 ==============================End of - Headers==============================
Has anyone used Ag or Sn sputter coatings to avoid low alpha peak pile ups? Au, Au/Pd and Pt are not really good when trying to find P presence. These are also a problem for W...etc.
All feedback is welcome.
gary g.
==============================Original Headers============================== 4, 17 -- From gary-at-gaugler.com Tue Feb 28 21:37:27 2006 4, 17 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 4, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k213bR58030335 4, 17 -- for {microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 21:37:27 -0600 4, 17 -- Received: (qmail 22755 invoked from network); 28 Feb 2006 19:37:27 -0800 4, 17 -- Received: by simscan 1.1.0 ppid: 22751, pid: 22752, t: 0.0956s 4, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 4, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 4, 17 -- by qsmtp1 with SMTP; 28 Feb 2006 19:37:27 -0800 4, 17 -- Message-Id: {6.2.3.4.2.20060228193437.0203cf58-at-mail.calweb.com} 4, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 4, 17 -- Date: Tue, 28 Feb 2006 19:37:27 -0800 4, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 4, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 4, 17 -- Subject: Low Z peak pileup 4, 17 -- Mime-Version: 1.0 4, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I wounder if there is a nanolithographic tool, other than SPM (STM or AFM), that can be used to scratch a line of 50-100 nm width on Si/SiO2 without using any masks?
Thanks
-- ********************************************************** Hany Ramadan Graduate student Chemistry department McMaster university, Hamilton, Ontario, Canada 905-525-9140 x: 26322 elsayeh-at-mcmaster.ca **********************************************************
==============================Original Headers============================== 5, 23 -- From ramadanhany-at-gmail.com Tue Feb 28 23:30:48 2006 5, 23 -- Received: from zproxy.gmail.com (zproxy.gmail.com [64.233.162.194]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k215Um3i015843 5, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 23:30:48 -0600 5, 23 -- Received: by zproxy.gmail.com with SMTP id 16so56927nzp 5, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 21:30:48 -0800 (PST) 5, 23 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 23 -- s=beta; d=gmail.com; 5, 23 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 5, 23 -- b=bKvEb3c1DV5CF9ZczJfqYGZw3uP42H0Z6o33CDqN+o6l0tUkjAdSW748BUcIKW+Q3zvrZUbfmIFHTi9MrM5WoSyKZyf65vI3kDHqaOWH8pNjIobRcm7oNYrS4Ne3YWa/qy8NvC3m+WPK8GVDmnYnysY2EpMxxPQXz4ddjlI6Qlc= 5, 23 -- Received: by 10.65.105.11 with SMTP id h11mr748542qbm; 5, 23 -- Tue, 28 Feb 2006 21:30:47 -0800 (PST) 5, 23 -- Received: by 10.65.159.14 with HTTP; Tue, 28 Feb 2006 21:30:47 -0800 (PST) 5, 23 -- Message-ID: {8d8ce5a30602282130o75939076v83d5bccfc394c07d-at-mail.gmail.com} 5, 23 -- Date: Wed, 1 Mar 2006 00:30:47 -0500 5, 23 -- From: "Hany Ramadan" {ramadanhany-at-gmail.com} 5, 23 -- To: Microscopy-at-microscopy.com 5, 23 -- Subject: nanolithography on SiO2 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; charset=ISO-8859-1 5, 23 -- Content-Disposition: inline 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k215Um3i015843 ==============================End of - Headers==============================
My wife had what appears to be a similar problem after a probably not so routine injection in the early 1980s. The injection was for protection from anthrax, which ironically in the end she never worked with. Although this has perhaps similarity with 'Gulf war Syndrome' 20 years later, her reaction to the injection seems very unlikely to be due the 'pathogen', more likely the carrier medium. She got an intense reaction at the site within 24 hours that remained very painful (couldn't be touched|) for a year or so, although the pain became intense and then reduced a little over weekly cycles. We don't think anything like steroid injections were tried as my wife certainly wasn't keen on any more needles, but it is a while ago now and she can't remember all the minor details (they must be in her notes).
As she describes below the only thing that got rid of it was a fairly major operation to remove the entire reaction site. It caused so much pain, although it got very bad and then a bit better in the weekly cycles, that she insisted on the operation to excise the area. As my wife mentioned, the first operation didn't remove enough of the inflamed area and the second probably removed slightly more than was necessary. However it was very successful in the end. She got no compensation or anything even though it was a work related injury, as nothing was really found (still seems a bit like Gulf War Syndrome) - the 'lump' was removed in a non-painful part of the cycle so there was no useful microscopy.list.server related histology. At the time such reactions seemed rare and little was known about them - it didn't even have a name (now probably all sorts of similar conditions are bundled under the 'macrophagic myofasciitis' label).
She had a minor complication due the resulting very large 6 inch vertical scar at the top of her left arm being 'wired' after the operation. A visiting locum doctor told her to change the rest position of the arm from that advised by the surgeons and it badly split the wound's stitching. The area of her arm now has a large sensitive scar (from the operation scar tissue) that hurts her if anyone pokes it and occasionally she gets the odd tingle from the old area near where the injection site was (probably scar tissue again), but nothing like she was suffering previously. Her arm clearly has a large dip where so much muscle tissue was removed, but it doesn't notice now she's older (53), although she tends to wear short sleeves rather than tank tops. Sunburn irritates it was well. However in comparison with the pain she suffered for a year or so when the inflamed area was present, she considers the operation was a complete success. This was all back in 1982, and medical staff here were non-plussed by the injection reaction at the time. She didn't get any systemic problems she was aware of, just around the injection site (although that was so painful it may have masked other symptoms I suppose). It was quite a drastic cure but it worked, and of course she went on to marry a great guy and have wonderful children while still juggling a successful career etc....
My wife's comments :
"They didn't treat it with anything. The only other cases I came across then (as it was pre-internet) were either allergy to the aluminium used in preparing the inoculum or in the 3rd world women reacted to the short chains of an 'oil' in the adjuvant which normally used long-chain molecules but not the inoculum itself.
Whatever it was, it was as instant as the day after the injection I could not lift or touch my arm.
Mine flared up every 2 weeks eventually like clockwork.
The first local excision did work but because they cut it out when it was 'quiet' they could not discern where it was. I told them they had cut it in half but of course they didn't believe me. So when it re-appeared on schedule 2 weeks later they decided to cut a huge amount out just to be sure. If they had done the 2nd op first it would not have needed to be so drastic but still sizeable."
I hope this is of some help.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
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==============================Original Headers============================== 16, 28 -- From keith.morris-at-ucl.ac.uk Wed Mar 1 04:17:37 2006 16, 28 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) 16, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21AHZEt004779 16, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 04:17:36 -0600 16, 28 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 16, 28 -- by vscani-d.ucl.ac.uk with smtp (Exim 4.51) 16, 28 -- id 1FEOOh-0007P3-Ns; Wed, 01 Mar 2006 10:17:31 +0000 16, 28 -- Message-ID: {003f01c63d19$3f7b26b0$7b865290-at-keithhigrade} 16, 28 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 16, 28 -- To: {Microscopy-at-microscopy.com} 16, 28 -- Cc: {W.Muss-at-salk.at} 16, 28 -- References: {200602271813.k1RID3wk015939-at-ns.microscopy.com} 16, 28 -- Subject: Re: [Microscopy] Vaccination complications: macrophagic myofasciitis 16, 28 -- Date: Wed, 1 Mar 2006 10:16:47 -0000 16, 28 -- MIME-Version: 1.0 16, 28 -- Content-Type: text/plain; 16, 28 -- format=flowed; 16, 28 -- charset="iso-8859-1"; 16, 28 -- reply-type=original 16, 28 -- Content-Transfer-Encoding: 7bit 16, 28 -- X-Priority: 3 16, 28 -- X-MSMail-Priority: Normal 16, 28 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 16, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 16, 28 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 16, 28 -- X-UCL-MailScanner: Found to be clean 16, 28 -- X-UCL-MailScanner-SpamCheck: 16, 28 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk ==============================End of - Headers==============================
I've used sputtered Ag on one occasion with some success. The only catch was that after sputtering, the samples went immediately into the scope, and once they were removed from the scope were not analyzed again. I didn't do any experiments to prove this, but I assumed the Ag would oxidize rather quickly. The Ag didn't do as good of a job of dissipating surface charge on the sample as Au would have, but it was adequate for the work I was doing.
Cheers, Bryan Bandli Research Microscopist MVA Scientific Consultants
gary-at-gaugler.com wrote:
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==============================Original Headers============================== 6, 19 -- From bbandli-at-mvainc.com Wed Mar 1 08:13:24 2006 6, 19 -- Received: from smtp05.gnvlscdb.sys.nuvox.net (smtp.nuvox.net [64.89.70.9]) 6, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21EDNut017324 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 08:13:24 -0600 6, 19 -- Received: from [192.168.1.70] (216.215.228.34.nw.nuvox.net [216.215.228.34]) 6, 19 -- by smtp05.gnvlscdb.sys.nuvox.net (8.12.11/8.12.11) with ESMTP id k21EDXHQ015464 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 09:13:33 -0500 6, 19 -- Message-ID: {4405AB92.6050208-at-mvainc.com} 6, 19 -- Date: Wed, 01 Mar 2006 09:11:30 -0500 6, 19 -- From: bbandli {bbandli-at-mvainc.com} 6, 19 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- MIME-Version: 1.0 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- Subject: Re: [Microscopy] Low Z peak pileup 6, 19 -- References: {200603010337.k213btxx031706-at-ns.microscopy.com} 6, 19 -- In-Reply-To: {200603010337.k213btxx031706-at-ns.microscopy.com} 6, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I sometimes use chromium or nickel to coat biological samples for EDS analysis. Cr can either be evaporated from Cr chips in vacuum evaporator, or sputtered from Cr target in sputter coater.The K-shell x-rays don't overlap any elements of biological interest and the L-shell is very low at about 0.57 KeV. Nickel L-shell is at about 0.89 KeV.
My experience with carbon coating is that it is not very good at eleiminating charging on highly insulating biological samples (critical point dried, freeze-dried) and also not being a "metal" is not a good source of secondary electrons for imaging. Bot Ni and Cr are very effective at eliminating charging and make for stable secondary electron images for capturing good images of what you are doing EDS analysis on.
Having said that, I shall attempt carbon coating today on biological sample to compare with EDS done on Cr coated sample a few weeks ago, so see if detectibility of trace amounts of Cu and Zn is improved.
Gib -- Gilbert (Gib) Ahlstrand, Electron Microscopist Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
-------- Original Message --------
Dear Keith, dear Phil (Oshel), dear Robert (Darby), Dear Diane (Ciaburry), dear Diane (Miller), dear listers, apologize for my perhaps rel. late reply to all kind opinions and/or messages I got concerning the header } Vaccination complications: macrophagic myofasciitis { (no other messages received except via MSA-Listserver).
I would like to thank you for your input and interesting/valuable comments, now knowing that there is not much new concerning an efficient treatment.
Yes, I have found also the reference on } one successful treatment { by a 2 years medication with steroids and azathioprine (Fischer,Reimann,Schroeder: Dtsch Med Wochenschr. 2003 Oct 31;128(44):2305-8) but in that case perhaps the myophagic "reaction" was not initiated by aluminium and therefore might have been caused by another constituent/circumstance .....the patient in our case unequivocally had Al in the macrophages' cytoplasm (as confirmed not only by typical EM-micrographs but now also by EDX and EELS) received corticosteroid treatment for appr. half a year without any improvement.
Chelating therapy for aluminium in human seems to be a little bit complicated perhaps, as I have seen from a lit. search, but perhaps I have located ONE person at Mount Sinai School of Medicine some minutes before. If you like, I'd send info directly / off Listserver to you provided that communication thread can be verified.
Also, I shall contact you personally within 24 h......(:-)) Have a nice and beautiful - great- day,
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Dear Wolgang,
My wife had what appears to be a similar problem after a probably not so routine injection in the early 1980s. The injection was for protection from anthrax, which ironically in the end she never worked with. Although this has perhaps similarity with 'Gulf war Syndrome' 20 years later, her reaction to the injection seems very unlikely to be due the 'pathogen', more likely the carrier medium. She got an intense reaction at the site within 24 hours that remained very painful (couldn't be touched|) for a year or so, although the pain became intense and then reduced a little over weekly cycles. We don't think anything like steroid injections were tried as my wife certainly wasn't keen on any more needles, but it is a while ago now and she can't remember all the minor details (they must be in her notes).
As she describes below the only thing that got rid of it was a fairly major operation to remove the entire reaction site. It caused so much pain, although it got very bad and then a bit better in the weekly cycles, that she insisted on the operation to excise the area. As my wife mentioned, the first operation didn't remove enough of the inflamed area and the second probably removed slightly more than was necessary. However it was very successful in the end. She got no compensation or anything even though it was a work related injury, as nothing was really found (still seems a bit like Gulf War Syndrome) - the 'lump' was removed in a non-painful part of the cycle so there was no useful microscopy.list.server related histology. At the time such reactions seemed rare and little was known about them - it didn't even have a name (now probably all sorts of similar conditions are bundled under the 'macrophagic myofasciitis' label).
She had a minor complication due the resulting very large 6 inch vertical scar at the top of her left arm being 'wired' after the operation. A visiting locum doctor told her to change the rest position of the arm from that advised by the surgeons and it badly split the wound's stitching. The area of her arm now has a large sensitive scar (from the operation scar tissue) that hurts her if anyone pokes it and occasionally she gets the odd tingle from the old area near where the injection site was (probably scar tissue again), but nothing like she was suffering previously. Her arm clearly has a large dip where so much muscle tissue was removed, but it doesn't notice now she's older (53), although she tends to wear short sleeves rather than tank tops. Sunburn irritates it was well. However in comparison with the pain she suffered for a year or so when the inflamed area was present, she considers the operation was a complete success. This was all back in 1982, and medical staff here were non-plussed by the injection reaction at the time. She didn't get any systemic problems she was aware of, just around the injection site (although that was so painful it may have masked other symptoms I suppose). It was quite a drastic cure but it worked, and of course she went on to marry a great guy and have wonderful children while still juggling a successful career etc....
My wife's comments :
"They didn't treat it with anything. The only other cases I came across then (as it was pre-internet) were either allergy to the aluminium used in preparing the inoculum or in the 3rd world women reacted to the short chains of an 'oil' in the adjuvant which normally used long-chain molecules but not the inoculum itself.
Whatever it was, it was as instant as the day after the injection I could not lift or touch my arm.
Mine flared up every 2 weeks eventually like clockwork.
The first local excision did work but because they cut it out when it was 'quiet' they could not discern where it was. I told them they had cut it in half but of course they didn't believe me. So when it re-appeared on schedule 2 weeks later they decided to cut a huge amount out just to be sure. If they had done the 2nd op first it would not have needed to be so drastic but still sizeable."
I hope this is of some help.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
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==============================Original Headers============================== 16, 28 -- From keith.morris-at-ucl.ac.uk Wed Mar 1 04:17:37 2006 16, 28 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) 16, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21AHZEt004779 16, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 04:17:36 -0600 16, 28 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 16, 28 -- by vscani-d.ucl.ac.uk with smtp (Exim 4.51) 16, 28 -- id 1FEOOh-0007P3-Ns; Wed, 01 Mar 2006 10:17:31 +0000 16, 28 -- Message-ID: {003f01c63d19$3f7b26b0$7b865290-at-keithhigrade} 16, 28 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 16, 28 -- To: {Microscopy-at-microscopy.com} 16, 28 -- Cc: {W.Muss-at-salk.at} 16, 28 -- References: {200602271813.k1RID3wk015939-at-ns.microscopy.com} 16, 28 -- Subject: Re: [Microscopy] Vaccination complications: macrophagic myofasciitis 16, 28 -- Date: Wed, 1 Mar 2006 10:16:47 -0000 16, 28 -- MIME-Version: 1.0 16, 28 -- Content-Type: text/plain; 16, 28 -- format=flowed; 16, 28 -- charset="iso-8859-1"; 16, 28 -- reply-type=original 16, 28 -- Content-Transfer-Encoding: 7bit 16, 28 -- X-Priority: 3 16, 28 -- X-MSMail-Priority: Normal 16, 28 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 16, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 16, 28 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 16, 28 -- X-UCL-MailScanner: Found to be clean 16, 28 -- X-UCL-MailScanner-SpamCheck: 16, 28 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk ==============================End of - Headers==============================
==============================Original Headers============================== 31, 28 -- From W.Muss-at-salk.at Wed Mar 1 10:41:27 2006 31, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 31, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21GfQch014880 31, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 10:41:26 -0600 31, 28 -- Received: from localhost (localhost [127.0.0.1]) 31, 28 -- by hermes.lks.at (Postfix) with ESMTP id CF10C5A9020; 31, 28 -- Wed, 1 Mar 2006 17:41:23 +0100 (CET) 31, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 31, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 31, 28 -- with ESMTP id 60933-01; Wed, 1 Mar 2006 17:41:23 +0100 (CET) 31, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 31, 28 -- by hermes.lks.at (Postfix) with SMTP id 67DC15A900A; 31, 28 -- Wed, 1 Mar 2006 17:41:23 +0100 (CET) 31, 28 -- Received: by localhost with Microsoft MAPI; Wed, 1 Mar 2006 17:41:10 +0100 31, 28 -- Message-ID: {01C63D57.53DD50E0.W.Muss-at-salk.at} 31, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 31, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 31, 28 -- To: "'keith.morris-at-ucl.ac.uk'" {keith.morris-at-ucl.ac.uk} 31, 28 -- Cc: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 31, 28 -- Subject: [Microscopy] Re(Summ): Vaccination complications: macrophagic myofasciitis 31, 28 -- Date: Wed, 1 Mar 2006 17:41:09 +0100 31, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 31, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 31, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 31, 28 -- MIME-Version: 1.0 31, 28 -- Content-Type: text/plain; charset="us-ascii" 31, 28 -- Content-Transfer-Encoding: 7bit 31, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Dear Hong, perhaps this citation could be also of value for your studies: (indeed I too was not able to find some better technical solution than : silver sulphide reaction(s), Gorm Danscher's methods, and Silver enhancement as Rick Powell formulated in his recent mail.
All best wishes Wolfgang Muss Salzburg/Austria
copied from HISTONET-Listserver to be found at: http://www.histosearch.com/histonet/May01/RE.Manganesestaining.html RE: Manganese staining
Roy Ellis {roy.ellis-at-imvs.sa.gov.au}
Beth, The following reference to manganese is found in 'Theory and strategy in histochemistry' edited by Hans Lyon and published by Springer-Verlag. After treating a section with benzothiazolylazonaphthol, manganese deposits will stain blue. The method is not specific as other metals, namely cadmium, zinc and nickel also stain blue. Post-differentiation with 1% oxine (8-hydroxyquinoline) in 75% ethanol turned tissue cadmium from blue to red but zinc remained blue. Regards Roy Ellis mailto:roy.ellis-at-imvs.sa.gov.au }
-----Original Message----- } From: Histo-Scientific Research Laboratories } [mailto:histosci-at-shentel.net] } Sent: Thursday, 3 May 2001 21:38 } To: Linda McGraf } Subject: Manganese staining } } } Dear Histonetters, } } Has anyone ever heard of staining for localization of manganese in animal } tissue? We have looked through our staining books and have come up with } nothing. If anyone has ever heard of such a procedure, please share any } info you may have. } } Thank you, } } Beth Poole } HSRL } beth-at-hsrl.org } (540)459-8211 } fax: (540)459-8217 } www.hsrl.org } 137 South Main Street } Woodstock, VA 22664
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America We have a PI here wants to localize manganese in tissue. Does anyone know a way of doing this without using X-ray microanalysis? Thank you.
Hong Emory
==============================Original Headers============================== 4, 16 -- From hyi-at-emory.edu Tue Feb 28 15:56:35 2006 4, 16 -- Received: from sccrmhc14.comcast.net (sccrmhc14.comcast.net [63.240.77.84]) 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1SLuZKn032163 4, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Feb 2006 15:56:35 -0600 4, 16 -- Received: from [24.30.86.130] (c-24-30-86-130.hsd1.ga.comcast.net[24.30.86.130]) 4, 16 -- by comcast.net (sccrmhc14) with SMTP 4, 16 -- id {2006022821563401400ebg66e} ; Tue, 28 Feb 2006 21:56:34 +0000 4, 16 -- Mime-Version: 1.0 (Apple Message framework v622) 4, 16 -- Content-Transfer-Encoding: 7bit 4, 16 -- Message-Id: {e81b3f049cea790cdb8341be7e80af1a-at-emory.edu} 4, 16 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 16 -- To: Microscopy-at-microscopy.com 4, 16 -- From: Hong Yi {hyi-at-emory.edu} 4, 16 -- Subject: Manganese 4, 16 -- Date: Tue, 28 Feb 2006 16:56:38 -0500 4, 16 -- X-Mailer: Apple Mail (2.622) ==============================End of - Headers==============================
==============================Original Headers============================== 15, 28 -- From W.Muss-at-salk.at Wed Mar 1 10:45:31 2006 15, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 15, 28 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21GjSok015853 15, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 10:45:29 -0600 15, 28 -- Received: from localhost (localhost [127.0.0.1]) 15, 28 -- by hermes.lks.at (Postfix) with ESMTP id 5EE5C5A9044; 15, 28 -- Wed, 1 Mar 2006 17:45:23 +0100 (CET) 15, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 15, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 15, 28 -- with ESMTP id 61136-01; Wed, 1 Mar 2006 17:45:22 +0100 (CET) 15, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 15, 28 -- by hermes.lks.at (Postfix) with SMTP id EE1B65A9041; 15, 28 -- Wed, 1 Mar 2006 17:45:22 +0100 (CET) 15, 28 -- Received: by localhost with Microsoft MAPI; Wed, 1 Mar 2006 17:45:10 +0100 15, 28 -- Message-ID: {01C63D57.E2BB1B80.W.Muss-at-salk.at} 15, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 15, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 15, 28 -- To: "'hyi-at-emory.edu'" {hyi-at-emory.edu} 15, 28 -- Cc: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 15, 28 -- Subject: AW: [Microscopy] Manganese 15, 28 -- Date: Wed, 1 Mar 2006 17:45:09 +0100 15, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 15, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 15, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 15, 28 -- MIME-Version: 1.0 15, 28 -- Content-Type: text/plain; charset="us-ascii" 15, 28 -- Content-Transfer-Encoding: 7bit 15, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Cr is known to oxidize quickly so I eliminated that one.
C would be good but the specimens may contain C.
I failed to list the elements I might see: B, C, O, F, Na, Mg, P, Si, Al, S, Cl, Ar, K, Ca, Fe, Co, Mo, In, Cu, Hf, W, Ga, As. Not all at once!
It is mostly the lower Zs that are a problem with low energy peaks around 2KeV.
Perhaps Pd alone is a good choice? This does not occur in organic dielectrics or other specimens.
gary g.
At 08:10 AM 3/1/2006, you wrote:
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==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Wed Mar 1 11:22:41 2006 13, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k21HMdss025692 13, 20 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 11:22:40 -0600 13, 20 -- Received: (qmail 21705 invoked from network); 1 Mar 2006 09:22:39 -0800 13, 20 -- Received: by simscan 1.1.0 ppid: 21702, pid: 21703, t: 0.1877s 13, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 20 -- by qsmtp3 with SMTP; 1 Mar 2006 09:22:39 -0800 13, 20 -- Message-Id: {6.2.3.4.2.20060301091618.0205cf68-at-mail.calweb.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 13, 20 -- Date: Wed, 01 Mar 2006 09:22:39 -0800 13, 20 -- To: ahlst007-at-umn.edu 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] RE: Low Z peak pileup] 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200603011610.k21GAN67009038-at-ns.microscopy.com} 13, 20 -- References: {200603011610.k21GAN67009038-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
How about TEM EELS? Does anyone out there know if this will work?
Aloha, Tina
_____________________________________________________________________________ Dear All: We have a PI here wants to localize manganese in tissue. Does anyone know a way of doing this without using X-ray microanalysis? Thank you.
Hong Emory
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * Biological Electron Microscope Facility * (808) 956-6251 * University of Hawaii at Manoa * * http://www.pbrc.hawaii.edu/bemf ****************************************************************************
==============================Original Headers============================== 8, 19 -- From tina-at-pbrc.hawaii.edu Wed Mar 1 12:52:51 2006 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 8, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21IqmNf013586 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 12:52:50 -0600 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k21Iqh0B022445 8, 19 -- (version=TLSv1/SSLv3 cipher=AES256-SHA bits=256 verify=NO) 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 08:52:44 -1000 (HST) 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k21IqgtI022442 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 08:52:43 -1000 (HST) 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 8, 19 -- Date: Wed, 1 Mar 2006 08:52:41 -1000 (HST) 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 8, 19 -- X-Sender: tina-at-halia 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 8, 19 -- Subject: AW: Manganese 8, 19 -- Message-ID: {Pine.GSO.4.21.0603010849240.22388-100000-at-halia} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
I have a student making C-Pt replicas. He shadows with Pt at 45 degree tilt with rotation. The protocol he has found then depoists C at 90 degrees to the source with rotation. He wants to know why he needs to change the angle of tilt to do the carbon. I could not really give him a good explanation. Can any of you help?
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21Jx6ko024033 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 13:59:07 -0600 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 4, 22 -- (authenticated bits=0) 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k21Jx5pL2826334 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 14:59:05 -0500 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 4, 22 -- X-Accept-Language: en-us, en 4, 22 -- MIME-Version: 1.0 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Replicas 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
When doing conventional replicas (static specimens, no rotation) the Pt evaporation is conducted at an angle to generate the shadows. The carbon is then evaporated at 90 degrees (directly above the specimen) to fill in the voids or gaps (shadows) generated during the Pt evaporation. This strengthens the replica.
In your case, you are rotating the specimen on a turntable in both cases. Nonetheless, even though the Pt evaporation is being carried out with rotation, you will still have some gaps (otherwise you would not see any shadows). The carbon (since it is being evaporated directly overhead) will not land in the same areas as the Pt but will more uniformly coat the specimen and fill in the shadows (gaps). Although it fills in the gaps, its low density does not interfere with the shadows generated by the Pt.
So, the reason for the different angles is to make the replica stronger by filling in any voids or gaps.
JB
} I have a student making C-Pt replicas. He shadows with Pt at 45 degree } tilt with rotation. The protocol he has found then depoists C at 90 } degrees to the source with rotation. He wants to know why he needs to } change the angle of tilt to do the carbon. I could not really give him } a good explanation. Can any of you help? } } Greg } } -- } Gregory W. Erdos, Ph.D, } Assistant Director, Biotechnology Program } Scientific Director, EM Core Lab } P.O. Box 118525 } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } Phone: 352-392-1295 } Fax: 352-846-0251 } } } ==============================Original Headers============================== } 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 } 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) } 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k21Jx6ko024033 } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } 13:59:07 -0600 } 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu } [10.227.60.63]) } 4, 22 -- (authenticated bits=0) } 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id } k21Jx5pL2826334 } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } 14:59:05 -0500 } 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} } 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 } 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} } 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) } 4, 22 -- X-Accept-Language: en-us, en } 4, 22 -- MIME-Version: 1.0 } 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" } {Microscopy-at-MSA.Microscopy.Com} } 4, 22 -- Subject: Replicas } 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 22 -- Content-Transfer-Encoding: 7bit } 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } 4, 22 -- X-Scanned-By: CNS Open Systems Group } (http://open-systems.ufl.edu/services/smtp-relay/) } 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group } (http://open-systems.ufl.edu/services/smtp-relay/) } ==============================End of - Headers==============================
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==============================Original Headers============================== 9, 18 -- From bozzola-at-siu.edu Wed Mar 1 14:28:03 2006 9, 18 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21KS3TZ029200 9, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 14:28:03 -0600 9, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 9, 18 -- by abbmta1.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k21KS1FY023612 9, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 14:28:02 -0600 (CST) 9, 18 -- Mime-Version: 1.0 9, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 9, 18 -- Message-Id: {p06110407c02bb21f9f99-at-[131.230.177.142]} 9, 18 -- In-Reply-To: {200603012010.k21KAXCC026054-at-ns.microscopy.com} 9, 18 -- References: {200603012010.k21KAXCC026054-at-ns.microscopy.com} 9, 18 -- Date: Wed, 1 Mar 2006 14:28:00 -0600 9, 18 -- To: Microscopy-at-msa.microscopy.com 9, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 9, 18 -- Subject: Re: [Microscopy] Replicas 9, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 9, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
We used to use Pt/C replicas to show surface relief. We would shadow a cellulose acetate surface replica with Pt/C at a small angle to the surface to highlight the surface texture, then deposit C normal to the surface to provide the support film. After the C film deposition the film was removed from the replica surface and mounted on a copper grid.
It is still useful when you want to see the height of features on a surface. If you know the shadow angle, you can easily calculate the height. I am not aware of doing sample rotation during the shadowing step, but then I'm a materials guy and am not familiar with some of the bio techniques.
Good luck, Henk
At 03:16 PM 03/01/06, gwe-at-ufl.edu wrote:
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==============================Original Headers============================== 10, 26 -- From colijn.1-at-osu.edu Wed Mar 1 14:54:52 2006 10, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21KspXt003731 10, 26 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 14:54:51 -0600 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 10, 26 -- id {01LZJO8BC9XS9V57H2-at-er6s1.eng.ohio-state.edu} for 10, 26 -- microscopy-at-microscopy.com; Wed, 01 Mar 2006 15:54:50 -0500 (EST) 10, 26 -- Received: from HOC1.ecr6.ohio-state.edu 10, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 10, 26 -- (PMDF V6.2-1x5 #31056) 10, 26 -- with ESMTPA id {01LZJO8AF6HG9VA6YH-at-er6s1.eng.ohio-state.edu} ; Wed, 10, 26 -- 01 Mar 2006 15:54:50 -0500 (EST) 10, 26 -- Date: Wed, 01 Mar 2006 15:54:47 -0500 10, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: [Microscopy] Replicas 10, 26 -- In-reply-to: {200603012016.k21KGm5p027267-at-ns.microscopy.com} 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- X-Sender: colijn-at-mail.ecr6.ohio-state.edu 10, 26 -- To: gwe-at-ufl.edu, Microscopy Listserver {microscopy-at-microscopy.com} 10, 26 -- Message-id: {6.1.0.6.2.20060301154803.02ee3940-at-mail.ecr6.ohio-state.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.0.6 10, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: {200603012016.k21KGm5p027267-at-ns.microscopy.com} ==============================End of - Headers==============================
Unless you change angles, the Pt and C will land in the same places. By changing the angle from 45 to 90 deg, the carbon covers pretty much everything (no shadows would be generated) since it comes straight down.
An interesting demonstration would be to mount an object on a turntable at the appropriate angles and use some colored spray paints.
} If the stage is tilted and rotating would the carbon not be able to } get in all the gaps? } } bozzola-at-siu.edu wrote: } } } } When doing conventional replicas (static specimens, no rotation) } } the Pt evaporation is conducted at an angle to generate the } } shadows. The carbon is then evaporated at 90 degrees (directly } } above the specimen) to fill in the voids or gaps (shadows) } } generated during the Pt evaporation. This strengthens the replica. } } } } In your case, you are rotating the specimen on a turntable in both } } cases. Nonetheless, even though the Pt evaporation is being carried } } out with rotation, you will still have some gaps (otherwise you } } would not see any shadows). The carbon (since it is being } } evaporated directly overhead) will not land in the same areas as } } the Pt but will more uniformly coat the specimen and fill in the } } shadows (gaps). Although it fills in the gaps, its low density does } } not interfere with the shadows generated by the Pt. } } } } So, the reason for the different angles is to make the replica } } stronger by filling in any voids or gaps. } } } } JB } } } } } I have a student making C-Pt replicas. He shadows with Pt at 45 degree } } } tilt with rotation. The protocol he has found then depoists C at 90 } } } degrees to the source with rotation. He wants to know why he needs to } } } change the angle of tilt to do the carbon. I could not really give him } } } a good explanation. Can any of you help? } } } } } } Greg } } } } } } -- } } } Gregory W. Erdos, Ph.D, } } } Assistant Director, Biotechnology Program } } } Scientific Director, EM Core Lab } } } P.O. Box 118525 } } } University of Florida } } } Gainesville, FL 32611 } } } gwe-at-ufl.edu } } } Phone: 352-392-1295 } } } Fax: 352-846-0251 } } } } } } } } } ==============================Original Headers============================== } } } 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 } } } 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) } } } 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } } k21Jx6ko024033 } } } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } } } 13:59:07 -0600 } } } 4, 22 -- Received: from [10.227.60.63] } } } (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) } } } 4, 22 -- (authenticated bits=0) } } } 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id } } } k21Jx5pL2826334 } } } 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 } } } 14:59:05 -0500 } } } 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} } } } 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 } } } 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} } } } 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) } } } 4, 22 -- X-Accept-Language: en-us, en } } } 4, 22 -- MIME-Version: 1.0 } } } 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" } } } {Microscopy-at-MSA.Microscopy.Com} } } } 4, 22 -- Subject: Replicas } } } 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } } 4, 22 -- Content-Transfer-Encoding: 7bit } } } 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } } } 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 } } } 4, 22 -- X-Scanned-By: CNS Open Systems Group } } } (http://open-systems.ufl.edu/services/smtp-relay/) } } } 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group } } } (http://open-systems.ufl.edu/services/smtp-relay/) } } } ==============================End of - Headers============================== } } } } } } } } } } } } } } -- } Gregory W. Erdos, Ph.D, } Assistant Director, Biotechnology Program } Scientific Director, EM Core Lab } P.O. Box 118525 } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } Phone: 352-392-1295 } Fax: 352-846-0251
==============================Original Headers============================== 7, 19 -- From bozzola-at-siu.edu Wed Mar 1 15:27:29 2006 7, 19 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 7, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21LRRIn013166 7, 19 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 15:27:28 -0600 7, 19 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 7, 19 -- by abbmta1.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k21LROtr015034 7, 19 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 1 Mar 2006 15:27:25 -0600 (CST) 7, 19 -- Mime-Version: 1.0 7, 19 -- X-Sender: bozzola-at-saluki-mail.siu.edu 7, 19 -- Message-Id: {p06110409c02bc11220b3-at-[131.230.177.142]} 7, 19 -- In-Reply-To: {44060A15.9080709-at-ufl.edu} 7, 19 -- References: {200603012037.k21KbwL4031717-at-ns.microscopy.com} 7, 19 -- {44060A15.9080709-at-ufl.edu} 7, 19 -- Date: Wed, 1 Mar 2006 15:27:23 -0600 7, 19 -- To: Microscopy-at-msa.microscopy.com 7, 19 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 7, 19 -- Subject: Re: [Microscopy] Re: Replicas 7, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 19 -- X-MASF: 0.00% ==============================End of - Headers==============================
Bozzola and Russell do a good job at diagrammatically showing why (if you want pictures - and well JB does a quick service to the list as well).
The Pt doesn't make a continuous film, even with rotation- esp if your samples have significant surface relief.
At 90° the carbon can be added uniformly and in a topographically neutral manner. Think about the carbon as the support for the Pt.
At least that's how I thought of the process. I had a luxury of having two sources: A point target with the Pt and a separate one for the carbon. And with the 90 degrees, I'd say drop the rotation maybe.
Does that help? When explaining something like this I always resort to drawing pictures.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: gwe-at-ufl.edu [mailto:gwe-at-ufl.edu] Sent: Wednesday, March 01, 2006 3:47 PM To: Williams, Geoffrey
I have a student making C-Pt replicas. He shadows with Pt at 45 degree tilt with rotation. The protocol he has found then depoists C at 90 degrees to the source with rotation. He wants to know why he needs to change the angle of tilt to do the carbon. I could not really give him a good explanation. Can any of you help?
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 4, 22 -- From gwe-at-ufl.edu Wed Mar 1 13:59:07 2006 4, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21Jx6ko024033 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 13:59:07 -0600 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 4, 22 -- (authenticated bits=0) 4, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k21Jx5pL2826334 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 1 Mar 2006 14:59:05 -0500 4, 22 -- Message-ID: {4405FD0A.40306-at-ufl.edu} 4, 22 -- Date: Wed, 01 Mar 2006 14:59:06 -0500 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 4, 22 -- X-Accept-Language: en-us, en 4, 22 -- MIME-Version: 1.0 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Replicas 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 4, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
==============================Original Headers============================== 16, 29 -- From Geoffrey_Williams-at-brown.edu Wed Mar 1 15:40:49 2006 16, 29 -- Received: from scorpio.services.brown.edu (scorpio.services.brown.edu [128.148.106.155]) 16, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21Lel6f017722 16, 29 -- for {Microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 15:40:48 -0600 16, 29 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 16, 29 -- by scorpio.services.brown.edu (Switch-3.1.7/Switch-3.1.7/) with ESMTP id k21LekQ1020973; 16, 29 -- Wed, 1 Mar 2006 16:40:47 -0500 (EST) 16, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 16, 29 -- Wed, 1 Mar 2006 16:40:46 -0500 16, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 16, 29 -- Content-class: urn:content-classes:message 16, 29 -- MIME-Version: 1.0 16, 29 -- Content-Type: text/plain; 16, 29 -- charset="iso-8859-1" 16, 29 -- Subject: RE: [Microscopy] Replicas 16, 29 -- Date: Wed, 1 Mar 2006 16:40:45 -0500 16, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F042303E5-at-MAIL1.AD.Brown.Edu} 16, 29 -- X-MS-Has-Attach: 16, 29 -- X-MS-TNEF-Correlator: 16, 29 -- Thread-Topic: [Microscopy] Replicas 16, 29 -- Thread-Index: AcY9cVbxc5ZMZm11QeSykRvDszMUyAABqVxg 16, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 16, 29 -- To: {gwe-at-ufl.edu} , {Microscopy-at-microscopy.com} 16, 29 -- X-OriginalArrivalTime: 01 Mar 2006 21:40:46.0700 (UTC) FILETIME=[CCBDD2C0:01C63D78] 16, 29 -- X-Brown-Proofpoint: Not Infected 16, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06021401 definitions=3.0.0-06030104 16, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06021401 definitions=3.0.0-06030104 16, 29 -- Content-Transfer-Encoding: 8bit 16, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k21Lel6f017722 ==============================End of - Headers==============================
I believe that they went with Pt/C to form small clusters for better resolution. It think that pure metals will nucleate in larger islands.
Henk
At 04:46 PM 03/01/06, you wrote:
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==============================Original Headers============================== 9, 26 -- From colijn.1-at-osu.edu Wed Mar 1 15:50:46 2006 9, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 9, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k21LokMR021322 9, 26 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 15:50:46 -0600 9, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 9, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 9, 26 -- id {01LZJQ6MUJUO9VIV3E-at-er6s1.eng.ohio-state.edu} for 9, 26 -- microscopy-at-microscopy.com; Wed, 01 Mar 2006 16:50:45 -0500 (EST) 9, 26 -- Received: from HOC1.ecr6.ohio-state.edu 9, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 9, 26 -- (PMDF V6.2-1x5 #31056) 9, 26 -- with ESMTPA id {01LZJQ6LTPQE9VK9JN-at-er6s1.eng.ohio-state.edu} ; Wed, 9, 26 -- 01 Mar 2006 16:50:44 -0500 (EST) 9, 26 -- Date: Wed, 01 Mar 2006 16:50:42 -0500 9, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 9, 26 -- Subject: Re: [Microscopy] Shadowing with evaporated metals 9, 26 -- In-reply-to: {200603012146.k21Lk3qW019641-at-ns.microscopy.com} 9, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 9, 26 -- X-Sender: colijn-at-mail.ecr6.ohio-state.edu 9, 26 -- To: bbandli-at-mvainc.com, microscopy-at-microscopy.com 9, 26 -- Message-id: {6.1.0.6.2.20060301164836.02d0dae0-at-mail.ecr6.ohio-state.edu} 9, 26 -- MIME-version: 1.0 9, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.0.6 9, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 9, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 9, 26 -- References: {200603012146.k21Lk3qW019641-at-ns.microscopy.com} ==============================End of - Headers==============================
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Title-Subject: [Filtered] fluorolume illuminator
Question: A colleague at a neighoring institution recently found what she described as a "fluorolume, old-style fluorescent illuminator". She asked me if I knew anything about how to operate it. I have never heard of a fluorolume and so am unable to help her right now. I would appreciate any responses regarding what it is, how it works, issues associated with it's use, etc...
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Title-Subject: [Filtered] SEM of bat embryos
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Title: SEM operation video
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Title: Magnification marker
Question: I have some TEM pictured that i got from scanned negative. these pictures have no magnification marker. my question is that how i can put a magnification marker on pictures when i have only negative and camrea length? thanks in advance
With an absorption edge ca 650 eV, Mn has a weak signal for EELS,. But it is doable, especially if the concentration is high enough. With freeze substituted cyanobacteria we get a cytosolic distribution of Mn in EELS ESI images.
Howard
On Mar 1, 2006, at 1:11 PM, tina-at-pbrc.hawaii.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ } 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 Hong- } } How about TEM EELS? Does anyone out there know if this will work? } } Aloha, } Tina } } ______________________________________________________________________ } _______ } Dear All: } We have a PI here wants to localize manganese in tissue. Does anyone } know a way of doing this without using X-ray microanalysis? Thank } you. } } Hong } Emory } } } ********************************************************************** } ****** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu } * Biological Electron Microscope Facility * (808) 956-6251 } * University of Hawaii at Manoa * } * http://www.pbrc.hawaii.edu/bemf } ********************************************************************** } ****** }
R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility Danforth Plant Science Center 975 N. Warson Rd. St. Louis, MO 63132
ph 314-587-1261 fx 314-587-1361 cell 314-378-2409 rhberg-at-danforthcenter.org www.danforthcenter.org visit this educational resource: http://www.danforthcenter.org/Cells/
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I want to provide a different perspective on the use of replicas. High-resolution Pt/C shadowing and replication provided important insights into the size and shape of polymers beginning over 40 years ago. The first images of DNA molecules were made this way. However, in my opinion, this methodology has largely been supplanted by the use of Atomic Force Microscopy, both for direct height measurements (available in Pt/C replicas by measuring shadow lengths when the coating is deposited from one direction only) and for imaging molecular contours.
As an everyday example, consider that making a magnetic read and write head for a hard disk drive requires controlling the relative heights of several different regions to a tolerance of ca. 1 nm. AFM supports production by providing a rapid means of offline analysis, far faster and more precise than any replica method could be. In the biopolymer area, for more than 10 years, it has been relatively easy to prepare dispersions of molecules on smooth surfaces like mica for AFM imaging. We have done some of this work ourselves, and examples can be seen at:
Disclaimer: ASM provides analytical services using AFM and I benefit from increasing the demand for AFM data.
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
----- Original Message ----- From: gwe-at-ufl.edu To: donc-at-asmicro.com Sent: Wednesday, March 01, 2006 3:37 PM Subject: [a] [Microscopy] Replicas
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I have a student making C-Pt replicas. He shadows with Pt at 45 degree tilt with rotation. The protocol he has found then depoists C at 90 degrees to the source with rotation. He wants to know why he needs to change the angle of tilt to do the carbon. I could not really give him a good explanation. Can any of you help?
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
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==============================Original Headers============================== 24, 22 -- From donc-at-asmicro.com Wed Mar 1 18:42:03 2006 24, 22 -- Received: from smtp103.sbc.mail.re2.yahoo.com (smtp103.sbc.mail.re2.yahoo.com [68.142.229.102]) 24, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k220g1aL011199 24, 22 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 18:42:02 -0600 24, 22 -- Received: (qmail 96736 invoked from network); 2 Mar 2006 00:42:01 -0000 24, 22 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.57.205.80 with login) 24, 22 -- by smtp103.sbc.mail.re2.yahoo.com with SMTP; 2 Mar 2006 00:42:00 -0000 24, 22 -- Message-ID: {000001c63d91$beaf7580$c901a8c0-at-ASM11} 24, 22 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 24, 22 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 24, 22 -- To: {gwe-at-ufl.edu} , "Microscopy List" {microscopy-at-microscopy.com} 24, 22 -- References: {200603012037.k21Kb5TY031477-at-ns.microscopy.com} 24, 22 -- Subject: Re: [a] [Microscopy] Replicas 24, 22 -- Date: Wed, 1 Mar 2006 18:06:00 -0500 24, 22 -- MIME-Version: 1.0 24, 22 -- Content-Type: text/plain; 24, 22 -- charset="iso-8859-1" 24, 22 -- Content-Transfer-Encoding: 7bit 24, 22 -- X-Priority: 3 24, 22 -- X-MSMail-Priority: Normal 24, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 24, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
One of my users has problems getting a good grid with negatively stained viral particles. She floats the grid on the virus+stain droplet, picks up the grid and then dries by touching against filter paper. It sounds like a pretty standard procedure but she often found that the support film ("store-bought" carbon coated formvar on either 300 or 400 mesh copper grid) is broken after the procedure. And occasionally, almost every hole is torn. Is there any tricks to prevent this?
I have experienced similar broken film but it was in formvar coated slot grids. After picking up a group of ribbons, the support film broke when the grid is dried. I always thought that I was just careless during the handling causing the film to crack and eventually the surface tension tore the film completely. However, that would be unlikely for the 300 or 400 mesh grids which are supported by so many grid bars? Thanks for any advice.
-- Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519) The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
==============================Original Headers============================== 4, 19 -- From wpchan-at-u.washington.edu Wed Mar 1 19:57:44 2006 4, 19 -- Received: from mxout5.cac.washington.edu (mxout5.cac.washington.edu [140.142.32.135]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k221vgwE007728 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 19:57:43 -0600 4, 19 -- Received: from homer23.u.washington.edu (homer23.u.washington.edu [140.142.12.141]) 4, 19 -- by mxout5.cac.washington.edu (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id k221vfws006724 4, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 17:57:42 -0800 4, 19 -- Received: from localhost (wpchan-at-localhost) 4, 19 -- by homer23.u.washington.edu (8.13.5+UW05.10/8.13.5+Submit) with ESMTP id k221vf6I025767 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 Mar 2006 17:57:41 -0800 4, 19 -- Date: Wed, 1 Mar 2006 17:57:41 -0800 (PST) 4, 19 -- From: "W. Chan" {wpchan-at-u.washington.edu} 4, 19 -- To: microscopy-at-microscopy.com 4, 19 -- Subject: viral particles 4, 19 -- Message-ID: {Pine.LNX.4.64.0603011722450.24739-at-homer23.u.washington.edu} 4, 19 -- MIME-Version: 1.0 4, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 19 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
The previous responders are correct in that the carbon makes the support film and the heavy metal creates shadows of the surface structure. The heavy metal angle is really a variable and should not always be set at 45 degrees per some protocol. The shallower the surface structure, the lower the Pt dep angle. As was stated a picture here would be a big help, but think of a 100nm high step vs a 2 nm high step on an otherwise smooth substrate. From my experience 45 degrees would be OK to shadow the 100nm step into visibility (actually a bit high). A Pt dep angle of 10 to 15 degrees would be better for the 2 nm step--longer shadows. Whatever angle you use you can make a rough calculation of step height from the geometry of the Pt shadowing--assuming the replica stays flat vs. assuming a potato chip shape.
Aside from AFM imaging, a Pt-C replica will give better topographic resolution of extremely small steps on a flat surface where secondary electron SEM contrast is weak, IMHO better than a super-duper SEM . Single-atom high growth steps on a growing crystal surface for example.
There is another "lost art" replication method I would love to see someone perform and send me the images to display in Microscopy Today: (I no longer have access to an e.m. lab). Pull a carbon replica of a fairly rough surface. Do not apply a heavy metal shadow. Put the naked carbon film into a TEM at 100keV or so. Tilt the specimen as high as your goniometer stage allows--45 to 60 degrees is best. Image with a small objective aperture in the bright-field position allowing the unscattered main beam to pass. Find an interesting step or structure using the weak contrast available in this mode. Then slightly displace the objective aperture so that the bright part of the main beam is *just* outside the aperture (or tilt the beam leaving the aperture centered to obtain the same effect). In other words, you are making a dark-field image using the "tails" of the main beam. Refocus. The result should be a sharp, high contrast, positive image that looks like and has the resolution of a high quality SEM image.
Ron Anderson, Editor Microscopy Today
gwe-at-ufl.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a student making C-Pt replicas. He shadows with Pt at 45 degree } tilt with rotation. The protocol he has found then depoists C at 90 } degrees to the source with rotation. He wants to know why he needs to } change the angle of tilt to do the carbon. I could not really give him } a good explanation. Can any of you help? } } Greg } }
==============================Original Headers============================== 6, 19 -- From randerson20-at-tampabay.rr.com Wed Mar 1 20:31:55 2006 6, 19 -- Received: from ms-smtp-05.tampabay.rr.com (ms-smtp-05-smtplb.tampabay.rr.com [65.32.5.135]) 6, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k222Vtn8018668 6, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 1 Mar 2006 20:31:55 -0600 6, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 6, 19 -- by ms-smtp-05.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k22202W8004771; 6, 19 -- Wed, 1 Mar 2006 21:00:04 -0500 (EST) 6, 19 -- Message-ID: {4406519F.1020900-at-tampabay.rr.com} 6, 19 -- Date: Wed, 01 Mar 2006 20:59:59 -0500 6, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 6, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 6, 19 -- MIME-Version: 1.0 6, 19 -- To: gwe-at-ufl.edu, Listserver {Microscopy-at-Microscopy.Com} 6, 19 -- Subject: Re: [Microscopy] Replicas 6, 19 -- References: {200603012000.k21K0Qm4024277-at-ns.microscopy.com} 6, 19 -- In-Reply-To: {200603012000.k21K0Qm4024277-at-ns.microscopy.com} 6, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit 6, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
unfortunately I do not have any idea how to be of help with handling such an illumination system.....but my personal interest in the request lead to the following search results:
............... ".....The basic tungsten source was the AO Ortho-illuminator and the high pressure mercury arc source was the AO Fluorolume illuminator equipped with a Corning (filter..)....." Source: Interchangeable Tungsten Filament and Mercury Arc Illuminator Adaptation for the Interference Microscope E. W. Lowrance Transactions of the American Microscopical Society, Vol. 86, No. 2 (Apr., 1967) , pp. 223-224 This journal is licensed to JSTOR by American Microscopical Society ==} http://links.jstor.org/sici?sici=0003-0023(196704)86%3A2%3C223%3AITFAMA% 3E2.0.CO%3B2-W
Structural Changes During Bean Leaf Abscission Peter C. Scott, Lillian W. Miller, Barbara D. Webster, A. C. Leopold American Journal of Botany, Vol. 54, No. 6 (Jul., 1967) , pp. 730-734 This journal is licensed to JSTOR by American Microscopical Society
Other Citations: Reference: Cellular and Molecular Life Sciences (CMLS) Publisher: Birkhauser Basel ISSN: 1420-682X (Paper) 1420-9071 (Online) DOI: 10.1007/BF01985701 Issue: Volume 37, Number 8
Question: A colleague at a neighoring institution recently found what she described as a "fluorolume, old-style fluorescent illuminator". She asked me if I knew anything about how to operate it. I have never heard of a fluorolume and so am unable to help her right now. I would appreciate any responses regarding what it is, how it works, issues associated with it's use, etc...
I have usually found that coatings collapse when the virus still has a lot of extraneous material associated with it such as from faecal samples or a not very pure cell pellet. I assumed it was mainly a heating and charging effect because of the high levels of background organic material which will swamp the conducting capacity of the carbon and copper on the grid. You never normally see it on pure isolates such as T4 phage. The simplest answer might be to dilute the drop until the grid stops breaking or spin out as much of the background as possible.
Your slot grid problem can easily happen because of flexing of the grid but it may also be weakened if you just coat the flat slot grids with plastic. I usually bend them slightly upwards so the plastic stretched across the and slot can't be damaged when it dries out.
I apologise if you have thought of all of this, already.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: wpchan-at-u.washington.edu
Dear Reza,
as I understand your question, in my opinion it is not possible to "put a magnification marker on pictures when (...you....) have only negative and camera length".......yes, you COULD DO that....but without guarantee for a "real" magnification reference.
2 solutions (only one really will work properly):
Solution 1: "real magnification" You must know the original magnification of the (T)EM-system onto the "negative".... (i.e.: primary magnification of EM-system at the correct kV-setting [= 50, 80, 100 kV; normally a certification test of magnification should be included in delivery papers of an instrument and should not change until major repair like lifting column parts, altering major adjustments of lenses etc.] times camera factor [depending on camera system you use [ e.g. 35 mm different will be a camera factor compared to other imaging/negative formats] = primary magnification of structures ON the NEGATIVE.
OR, right from the beginning, you MUST KNOW the negative's magnification.
If you then enlarge (secondary magnification) by means of an ancillary enlarger system onto positive paper, you have to multiply primary magnification ON the NEGATIVE with the "factor" of your (secondary) enlarger....on a positive you then can draw bars with the apropriate length due to secondary enlargement, scanning the positive, you will have included the "real" and correct magnification of structure/image....
Solution two: this only will result in a very "approximatively" set magnification bar: if there is included a sructure of known "normal" length, width, diameter...etc you perhaps CAN DRAW a line indicating } ~ ?m { or so........
Real magnification out of an unknown magnified negative } times { known secondary enlargement (?camera length?) unfortunately results again in } unknown magnification {....
So at least you should search for the (primary) magnification of the negative......
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Title: Magnification marker
Question: I have some TEM pictured that i got from scanned negative. these pictures have no magnification marker. my question is that how i can put a magnification marker on pictures when i have only negative and camrea length? thanks in advance
For those of you who were curious as to why we were using rotary shadowing, I would refer you to the web page of Gary Borisy http://www.borisylab.northwestern.edu/pages/protocols/electmicrosc.html#metcoat
Look at Figs. 4,5,6,8 to see the result of such shadowing on cytoskeletal proteins.
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 5, 22 -- From gwe-at-ufl.edu Thu Mar 2 07:44:10 2006 5, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22Di9UC021977 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 2 Mar 2006 07:44:09 -0600 5, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 5, 22 -- (authenticated bits=0) 5, 22 -- by smtp.ufl.edu (8.13.4/8.13.4/2.5.1) with ESMTP id k22Di6iY4321324 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 2 Mar 2006 08:44:07 -0500 5, 22 -- Message-ID: {4406F6A8.9090601-at-ufl.edu} 5, 22 -- Date: Thu, 02 Mar 2006 08:44:08 -0500 5, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 5, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 5, 22 -- X-Accept-Language: en-us, en 5, 22 -- MIME-Version: 1.0 5, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- Subject: Replicas FYI 5, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 5, 22 -- X-UFL-Spam-Status: hits=-1.428, required=5, tests=BAYES_20 5, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 5, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Pang, When you say that she dries the grids by touching them to filter paper....is she actually touching the face of the grids, or wicking the excess fluid by touching the edge of the grid to the paper? This is a critical difference. Its all in the interpretation of a written protocol. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 21 -- From lcgould-at-med.cornell.edu Thu Mar 2 08:06:57 2006 1, 21 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22E6vlu026341 1, 21 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 08:06:57 -0600 1, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 21 -- by smtp-gw2.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k22E6s4U023189 1, 21 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 09:06:54 -0500 (EST) 1, 21 -- Received: from [140.251.48.23] by mpx1.med.cornell.edu 1, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 21 -- with ESMTP id {0IVI003IA77HES40-at-mpx1.med.cornell.edu} for 1, 21 -- microscopy-at-microscopy.com; Thu, 02 Mar 2006 09:06:54 -0500 (EST) 1, 21 -- Date: Thu, 02 Mar 2006 09:02:51 -0500 1, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 21 -- Subject: Re: [Microscopy] viral particles 1, 21 -- In-reply-to: {200603020217.k222H7BQ014221-at-ns.microscopy.com} 1, 21 -- To: wpchan-at-u.washington.edu, Microscopy Listserver {microscopy-at-microscopy.com} 1, 21 -- Message-id: {p06200700c02cab041a7d-at-[140.251.48.23]} 1, 21 -- MIME-version: 1.0 1, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 21 -- References: {200603020217.k222H7BQ014221-at-ns.microscopy.com} 1, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.03.02.052605 ==============================End of - Headers==============================
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Title-Subject: [Filtered] FET specification (Pre-amplifier 183 A)
Question: Dear all,
I'm working with a FRX Philips PV 9500 with a detector EDAX (Pre-amplifier 183 A). The beryllium window was replaced since it was broken, but the detector still doesnĄt work. After re-checking, I can conclude that the FET of the detector is broken (doesn't work). Then I need to know the technical specifications or data sheet of this FET.
Any help about specification would be appreciated.
Thanks,
TČc. Ppal Elbio MartĚnez SECEGRIN - CERIDE - CONICET G¸emes 3450 3000 - Santa Fe Argentina email: edmarti-at-ceride.gov.ar
Instead of floating the filmed grid on the virus/stain mix try attaching the grid to a grid stick that has adhesive applied. Or you can put a piece of double-sided scotch tape onto the edge of a glass microscope slide and attach the extreme edge of the grid to that.
Now apply the virus/stain drop and after the appropriate time remove the liquid by touching the edge of the grid with a piece of filter paper.
This provides much gentler handling of the support film than lifting the grid off a droplet of solution where the surface tension creates quite a pull on the film as the grid is lifted away.
Certainly, using this technique, the support film should not rupture even on a 200 mesh grid.
Good luck,
Ted Dunn
--- wpchan-at-u.washington.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi } } One of my users has problems getting a good grid } with negatively stained } viral particles. She floats the grid on the } virus+stain droplet, picks up } the grid and then dries by touching against filter } paper. It sounds like } a pretty standard procedure but she often found that } the support film } ("store-bought" carbon coated formvar on either 300 } or 400 mesh copper } grid) is broken after the procedure. And } occasionally, almost every hole } is torn. Is there any tricks to prevent this? } } I have experienced similar broken film but it was in } formvar coated slot } grids. After picking up a group of ribbons, the } support film broke when } the grid is dried. I always thought that I was just } careless during the } handling causing the film to crack and eventually } the surface tension tore } the film completely. However, that would be } unlikely for the 300 or 400 } mesh grids which are supported by so many grid bars? } Thanks for any } advice. } } -- } Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB } A087, 206-685-1519) } The Biology Imaging Facility } (http://staff.washington.edu/wpchan/if/) } } ==============================Original } Headers============================== } 4, 19 -- From wpchan-at-u.washington.edu Wed Mar 1 } 19:57:44 2006 } 4, 19 -- Received: from mxout5.cac.washington.edu } (mxout5.cac.washington.edu [140.142.32.135]) } 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k221vgwE007728 } 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 } Mar 2006 19:57:43 -0600 } 4, 19 -- Received: from homer23.u.washington.edu } (homer23.u.washington.edu [140.142.12.141]) } 4, 19 -- by mxout5.cac.washington.edu } (8.13.5+UW05.10/8.13.5+UW05.09) with ESMTP id } k221vfws006724 } 4, 19 -- (version=TLSv1/SSLv3 } cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 } Mar 2006 17:57:42 -0800 } 4, 19 -- Received: from localhost (wpchan-at-localhost) } 4, 19 -- by homer23.u.washington.edu } (8.13.5+UW05.10/8.13.5+Submit) with ESMTP id } k221vf6I025767 } 4, 19 -- for {microscopy-at-microscopy.com} ; Wed, 1 } Mar 2006 17:57:41 -0800 } 4, 19 -- Date: Wed, 1 Mar 2006 17:57:41 -0800 (PST) } 4, 19 -- From: "W. Chan" {wpchan-at-u.washington.edu} } 4, 19 -- To: microscopy-at-microscopy.com } 4, 19 -- Subject: viral particles } 4, 19 -- Message-ID: } {Pine.LNX.4.64.0603011722450.24739-at-homer23.u.washington.edu} } 4, 19 -- MIME-Version: 1.0 } 4, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; } format=flowed } 4, 19 -- X-Uwash-Spam: Gauge=IIIIIII, } Probability=7%, Report='__C230066_P5 0, __CT 0, } __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY } 0, __MIME_VERSION 0, __SANE_MSGID 0' } ==============================End of - } Headers============================== }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 11, 19 -- From drteddunne-at-yahoo.com Thu Mar 2 09:23:29 2006 11, 19 -- Received: from web33411.mail.mud.yahoo.com (web33411.mail.mud.yahoo.com [68.142.206.143]) 11, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k22FNSpZ012360 11, 19 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 09:23:29 -0600 11, 19 -- Received: (qmail 94792 invoked by uid 60001); 2 Mar 2006 15:23:28 -0000 11, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 19 -- s=s1024; d=yahoo.com; 11, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 11, 19 -- b=U2u1+/qPDDqTnvnvab7y+jwzrwPz1JMTPbDwXwNgBXqT2RpTGiGaNP3FizX7k/xCqlRdoplngnQEZcqPJYZ1hX+VtYUjeWNMrn1ADVQHfZv3MjLjocLVotuGR3ZWFnJqDYS2cIzSqtL/XkuEyRStnB5xeIp1OCl9LsNdWscuvlk= ; 11, 19 -- Message-ID: {20060302152328.94790.qmail-at-web33411.mail.mud.yahoo.com} 11, 19 -- Received: from [202.47.247.156] by web33411.mail.mud.yahoo.com via HTTP; Thu, 02 Mar 2006 07:23:28 PST 11, 19 -- Date: Thu, 2 Mar 2006 07:23:28 -0800 (PST) 11, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 11, 19 -- Subject: Re: [Microscopy] viral particles 11, 19 -- To: microscopy-at-microscopy.com 11, 19 -- In-Reply-To: {200603020311.k223Bp32031503-at-ns.microscopy.com} 11, 19 -- MIME-Version: 1.0 11, 19 -- Content-Type: text/plain; charset=iso-8859-1 11, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I have just gotten this message, but the message I sent had no attachments. Is there any way you can check this? Maybe I have a virus...
Thank you,
dj
On Thu, 2 Mar 2006 spamMfilter-at-microscopy.com wrote:
} Subject: REJECTED MAIL } X-Mailer: MicroscopyListSpam Mail Filter vNJZ-2005080908 } X-lewp: MicroscopyListSpam NAGS } } -------------------------------------------------------- } Your mail has been rejected for the following reason(s): } -------------------------------------------------------- } Other: } } Content-Type: multipart/alternative } } An Email Attachment was detected with your message!!! } The Microscopy Listserver will not accept any ATTACHMENTS as a safety measure. } Suspect or Possibly an Unregistered User Address: dljones-at-bestweb.net } } Site match: verizon.net } --------------------------------------------------------------------- } If you have a legitimate reason to contact this SITE, you may get your } mail through the filter by FORWARDing this Email together with } all the error messages and header linesto the Address: } } } } MicroscopyListSpamFilter-at-Microscopy.Com: } } --------------------------------------------------------------------- } } } MicroscopyList Email Filter vNJZ-2005080908 - Your Friendly Neighborhood SysOp } } The text of the rejected email follows: } ********************************************************************* } } This message is in MIME format. The first part should be readable text, } while the remaining parts are likely unreadable without MIME-aware tools. } } --6400082-538-1141314083=:1576 } Content-Type: TEXT/PLAIN; charset=X-UNKNOWN; format=flowed } Content-Transfer-Encoding: QUOTED-PRINTABLE } } Giovanni, } } I do have a video for an Amray 1100 SEM. I haven't looked at it in a long t= } ime=20 } but I believe it has a lot of fundamentals of electron microscopy and EDS= } =20 } operation that is independent of instrument, although the AMR 1100 is quite= } =20 } similar to the AMR 1000 you are asking for. In fact, if you could send me a= } =20 } photo of the main control panel, I could probably indicate the differences = } and=20 } similarities in the two. } } I don't recall if it has much about sample preperation, but if it did, it w= } ould=20 } likely be aimed at metallurgy and not biological samples, which are probabl= } y=20 } what you would prefer. But I'm sure you can more easily find sample prep in= } fo=20 } focused towards biology fairly easily. } } The video I have is neither in VHS or DVD, it is in Video 8. I will have to= } =20 } figure out how to convert these tapes. } } I would be willing to copy them and send them to you for the base cost of= } =20 } materials and shipping. } } Let me know how you would like to proceed. } } dj } } On Wed, 1 Mar 2006 exploratorium-at-tiscali.it wrote: } } --| Email: exploratorium-at-tiscali.it } --| Name: giovanni de caro } --| } --| Organization: associazione luigi montalb=DA } --| } --| Education: 9-12th Grade High School } --| } --| Location: Campobasso, Italy } --| } --| Title: SEM operation video } --| } --| Question: I am looking for a video (VHS or DVD) dealing with the practica= } l=20 } --| operation of an older SEM. We are going to restart and operate an AMR 100= } 0 SEM=20 } --| in our natural science museum for youngsters in Italy=20 } --| (http://web.tiscali.it/exploratorium). Of course I do not expect a docume= } ntary=20 } --| ilustrating THIS specific instrument, I would be more than happy of somet= } hing=20 } --| showing the operation of this calss of instruments. I also would like to = } have=20 } --| a video on SEM specimen preparation. Thank you for your kind help. } --6400082-538-1141314083=:1576-- } } } ==============================Original Headers============================== } 12, 25 -- From dljones-at-bestweb.net Thu Mar 2 09:31:56 2006 } 12, 25 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) } 12, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22FVtKf014572 } 12, 25 -- for |--Microscopy-at-microscopy.com--|; Thu, 2 Mar 2006 09:31:56 -0600 } 12, 25 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) } 12, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id C1DB41CCF0; } 12, 25 -- Thu, 2 Mar 2006 10:31:54 -0500 (EST) } 12, 25 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) } 12, 25 -- by smtp2.bestweb.net (Postfix) with ESMTP id 400E61CCCB; } 12, 25 -- Thu, 2 Mar 2006 10:31:54 -0500 (EST) } 12, 25 -- Date: Thu, 2 Mar 2006 10:41:23 -0500 (Eastern Standard Time) } 12, 25 -- From: "David L. Jones" |--dljones-at-bestweb.net--| } 12, 25 -- To: exploratorium-at-tiscali.it } 12, 25 -- cc: Microscopy-at-microscopy.com } 12, 25 -- Subject: Re: [Microscopy] AskAMicroscopist: SEM operation video } 12, 25 -- In-Reply-To: |--200603020141.k221fcTU001827-at-ns.microscopy.com--| } 12, 25 -- Message-ID: |--Pine.WNT.4.62.0603021029330.1576-at-dlj--| } 12, 25 -- References: |--200603020141.k221fcTU001827-at-ns.microscopy.com--| } 12, 25 -- X-X-Sender: dljones-at-pop-croton.bestweb.net } 12, 25 -- MIME-Version: 1.0 } 12, 25 -- Content-Type: MULTIPART/MIXED; BOUNDARY="6400082-538-1141314083=:1576" } 12, 25 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net } 12, 25 -- X-Spam-Level: } 12, 25 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed } 12, 25 -- version=3.0.2 } ==============================End of - Headers============================== }
==============================Original Headers============================== 7, 24 -- From dljones-at-bestweb.net Thu Mar 2 09:39:53 2006 7, 24 -- Received: from smtp2.bestweb.net (smtp2-2.bestweb.net [209.94.103.46]) 7, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22FdqZQ016721 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 09:39:52 -0600 7, 24 -- Received: from smtp2.bestweb.net (localhost [127.0.0.1]) 7, 24 -- by smtp2.bestweb.net (Postfix) with ESMTP id DC68A1CCC3 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 10:39:51 -0500 (EST) 7, 24 -- Received: from dlj (pool-71-249-9-96.nycmny.east.verizon.net [71.249.9.96]) 7, 24 -- by smtp2.bestweb.net (Postfix) with ESMTP id 570F21CC37 7, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 10:39:51 -0500 (EST) 7, 24 -- Date: Thu, 2 Mar 2006 10:49:20 -0500 (Eastern Standard Time) 7, 24 -- From: "David L. Jones" {dljones-at-bestweb.net} 7, 24 -- To: Microscopy-at-microscopy.com 7, 24 -- Subject: Re: AskAMicroscopist: SEM operation video 7, 24 -- In-Reply-To: {200603021531.k22FVwuC014586-at-ns.microscopy.com} 7, 24 -- Message-ID: {Pine.WNT.4.62.0603021048000.1576-at-dlj} 7, 24 -- References: {200603021531.k22FVwuC014586-at-ns.microscopy.com} 7, 24 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 7, 24 -- MIME-Version: 1.0 7, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 7, 24 -- X-Spam-Checker-Version: SpamAssassin 3.0.2 (2004-11-16) on smtp2-1.bestweb.net 7, 24 -- X-Spam-Level: 7, 24 -- X-Spam-Status: No, score=-2.8 required=5.0 tests=ALL_TRUSTED autolearn=failed 7, 24 -- version=3.0.2 ==============================End of - Headers==============================
If the student is looking for macromolecules you actually need an angle of 8 to10 degrees for Pt and 90 degrees for Carbon. If he uses high angles like 45 he will not see anything. Those high angles were used for big stuff like Bacteria and some of the larger viruses like TMV. I have several protocols that I can supply off line.
Best, Al Coritz, Technical Engineer Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- X-from: {gwe-at-ufl.edu} To: {Sampleprep-at-earthlink.net} Sent: Wednesday, March 01, 2006 3:04 PM
Colleagues, As I have not seen a post on the subject of Rob Apkarian's accidental death yesterday I thought I would convey the sad news to the list. I did not know Rob well but we met and interacted at M&M meetings over many years. Rob had enthusiasm in abundance for electron microscopy and I will remember him as a "real character".
Regards
Chris
Christopher J Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.A04 UT Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-9039 Phone +1 214 648 2827 Fax +1 214 648 6408
==============================Original Headers============================== 5, 21 -- From christopher.gilpin-at-utsouthwestern.edu Thu Mar 2 15:16:25 2006 5, 21 -- Received: from swlx167.swmed.edu (swlx167.swmed.edu [199.165.152.167]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22LGO6o023123 5, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 15:16:24 -0600 5, 21 -- Message-Id: {200603022116.k22LGO6o023123-at-ns.microscopy.com} 5, 21 -- Received: from [129.112.148.58] (helo=cgdesktop) 5, 21 -- by swlx167.swmed.edu with esmtp (Exim 4.44) 5, 21 -- id 1FEv9s-0001YB-3p 5, 21 -- for Microscopy-at-microscopy.com; Thu, 02 Mar 2006 15:16:24 -0600 5, 21 -- From: "Christopher Gilpin" {christopher.gilpin-at-utsouthwestern.edu} 5, 21 -- To: {Microscopy-at-microscopy.com} 5, 21 -- Date: Thu, 2 Mar 2006 15:16:36 -0600 5, 21 -- MIME-Version: 1.0 5, 21 -- Content-Type: text/plain; 5, 21 -- charset="us-ascii" 5, 21 -- Content-Transfer-Encoding: 7bit 5, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 5, 21 -- Thread-Index: AcY+PpbOIka7Tyf2TCO7Qu1UmfNBBA== 5, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 5, 21 -- X-Scan-Signature: ab3f4c55490e4d672805895add812bc5 5, 21 -- Subject: Rob Apakrian passed away ==============================End of - Headers==============================
Thanks for the feedback on dealing with low eV peak pileup.
It looks like Ni and Pd are good options. Does anyone have experience with these and know if they oxidize?
Ni is 99.9% pure and is .062" thick. Pd is 99.5% pure and can be .004" or .008" thick. So I guess that Ni deposits faster than Pd. I recall the typical thickness for Au/Pd is about .01".
I don't think that just one target type will work for all specimens. That is OK. Changing targets is not all that big of a deal.
Thanks for the help.
gary g.
==============================Original Headers============================== 7, 17 -- From gary-at-gaugler.com Thu Mar 2 16:06:15 2006 7, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 7, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k22M6E0b000319 7, 17 -- for {microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 16:06:14 -0600 7, 17 -- Received: (qmail 23885 invoked from network); 2 Mar 2006 14:06:05 -0800 7, 17 -- Received: by simscan 1.1.0 ppid: 23882, pid: 23883, t: 0.1081s 7, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 7, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 7, 17 -- by qsmtp2 with SMTP; 2 Mar 2006 14:06:04 -0800 7, 17 -- Message-Id: {6.2.3.4.2.20060302135842.01ff6768-at-mail.calweb.com} 7, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 7, 17 -- Date: Thu, 02 Mar 2006 14:06:14 -0800 7, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 7, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 7, 17 -- Subject: Ni and Pd target material 7, 17 -- Mime-Version: 1.0 7, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
With deep sorrow, I am passing onto the list the tragic news of Dr. Rob Apkarian's Death, which occurred in Atlanta in the evening of February 28, 2006. Many of you may know Rob for his remarkable work in cryo-EM and high resolution SEM. Some of you may also have active collaborations with Rob. Rob was the director of Integrated Microscopy and Microanalytical Facility in Emory and a close colleague of mine in Emory microscopy research community. Rob was also an active member of the MSA leadership for many years.
Currently, I do not know about any plan regarding any religious service or funeral yet. But if anyone wishes to send a condolence card or flowers, I can help you get connected if you contact me off-line. Rob is survived by his wife who is also a member of Emory community.
Take care and live well.
Hong Emory EM
==============================Original Headers============================== 10, 18 -- From hyi-at-emory.edu Thu Mar 2 17:24:13 2006 10, 18 -- Received: from rwcrmhc11.comcast.net (rwcrmhc11.comcast.net [216.148.227.151]) 10, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k22NOCjx011488 10, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 17:24:12 -0600 10, 18 -- Received: from [24.30.86.130] (c-24-30-86-130.hsd1.ga.comcast.net[24.30.86.130]) 10, 18 -- by comcast.net (rwcrmhc11) with SMTP 10, 18 -- id {20060302232404m11009rnpie} ; Thu, 2 Mar 2006 23:24:08 +0000 10, 18 -- Mime-Version: 1.0 (Apple Message framework v622) 10, 18 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 18 -- Message-Id: {8cdae9f5fb505007537fe576bc1aeeb9-at-emory.edu} 10, 18 -- Cc: BusinessOffice-at-microscopy.org 10, 18 -- From: Hong Yi {hyi-at-emory.edu} 10, 18 -- Subject: (Microscopy) Dr. Apkarian 10, 18 -- Date: Thu, 2 Mar 2006 18:24:10 -0500 10, 18 -- To: Microscopy-at-microscopy.com 10, 18 -- X-Mailer: Apple Mail (2.622) 10, 18 -- Content-Transfer-Encoding: 8bit 10, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k22NOCjx011488 ==============================End of - Headers==============================
Thank you Chris and Hong for the posts about Rob's passing. A service for Rob is being planned for next week, the details have not been finalized. For those of you wishing to extend your sympathies to Rob's wife, you may contact me off-line as well.
He was a great research scientist, mentor, and a dear friend. I will miss him deeply.
Sincerely,
Elizabeth R. Wright
==============================Original Headers============================== 6, 20 -- From erwright-at-caltech.edu Thu Mar 2 18:16:08 2006 6, 20 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k230G6rA021415 6, 20 -- for {microscopy-at-Microscopy.Com} ; Thu, 2 Mar 2006 18:16:07 -0600 6, 20 -- Received: from localhost (water-dog [192.168.1.26]) 6, 20 -- by fire-ox-postvirus (Postfix) with ESMTP 6, 20 -- id 1598435E88; Thu, 2 Mar 2006 16:16:06 -0800 (PST) 6, 20 -- Received: from [192.168.157.114] (pix-1.caltech.edu [131.215.2.21]) 6, 20 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP 6, 20 -- id 112C535C32; Thu, 2 Mar 2006 16:16:05 -0800 (PST) 6, 20 -- Mime-Version: 1.0 (Apple Message framework v623) 6, 20 -- Content-Transfer-Encoding: 7bit 6, 20 -- Message-Id: {ee1d05f1abe4f266fe60b5bc3cb54fea-at-caltech.edu} 6, 20 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 6, 20 -- To: microscopy-at-Microscopy.Com, 3dem-at-ucsd.edu 6, 20 -- From: "Elizabeth R.Wright" {erwright-at-caltech.edu} 6, 20 -- Subject: Dr. Robert P. Apkarian 6, 20 -- Date: Thu, 2 Mar 2006 16:16:04 -0800 6, 20 -- X-Mailer: Apple Mail (2.623) 6, 20 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mariac-h-at-uniandes.edu.co) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 2, 2006 at 18:29:31 ---------------------------------------------------------------------------
Email: mariac-h-at-uniandes.edu.co Name: Maria Cristina Herrera
Organization: Universidad de Los Andes
Education: Graduate College
Location: Bogota, Colombia
Question: I got some SEM images of soils. I can identify some minerals but I am not sure about somethings that look like dehidrated roots. The scale of my images is 10 microns. The roots are longer than 10 microns and about 1 micron thick. I would like to know if roots can be this large and if there is somebody who can help me to define what are those things in my images..I can email my images. Thanks.
If the specimen is very small, and/or has low contrast (unstained), and can be deposited onto a formvar or carbon-coated grid, then the student could shadow the sample on-grid with Pt/C and omit the carbon altogether. The carbon layer is only important so that the replica doesn't distort or crack when it is floated off of the substrate. Because there is no extra carbon layer, there is a slight increase in contrast with an on-grid shadow vs a true replica. As mentioned in previous replies, small samples usually look best when shadowed at a low angle (10-15 deg. from horizontal). Furthermore, rotary shadowing usually looks best on fibers while fixed-angle shadowing looks best on globular/particulate specimens. And finally, there are less steps involved in making an on-grid shadow vs a replica.
Sorry if we've beat this topic to death,
Andrew Bowling The University of Texas at Austin
==============================Original Headers============================== 6, 17 -- From abowling-at-mail.utexas.edu Thu Mar 2 18:48:04 2006 6, 17 -- Received: from wb6-a.mail.utexas.edu (wb6-a.mail.utexas.edu [128.83.126.144]) 6, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k230m2nb028499 6, 17 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Mar 2006 18:48:02 -0600 6, 17 -- Received: (qmail 22244 invoked from network); 3 Mar 2006 00:48:01 -0000 6, 17 -- Received: from dhcp-146-6-151-204.biosci.utexas.edu (HELO ?146.6.151.204?) (abowling-at-146.6.151.204) 6, 17 -- by wb6.mail.utexas.edu with (RC4-MD5 encrypted) ESMTPSA; 3 Mar 2006 00:48:01 -0000 6, 17 -- Message-ID: {440791F4.2080606-at-mail.utexas.edu} 6, 17 -- Date: Thu, 02 Mar 2006 18:46:44 -0600 6, 17 -- From: Andrew Bowling {abowling-at-mail.utexas.edu} 6, 17 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 6, 17 -- X-Accept-Language: en-us, en 6, 17 -- MIME-Version: 1.0 6, 17 -- To: Microscopy-at-microscopy.com 6, 17 -- Subject: [Microscopy] Re: Replicas 6, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (andromeda_tm-at-libero.it) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 3, 2006 at 04:32:02 ---------------------------------------------------------------------------
Hi all, does someone know the protocol for staining with the ěEhrlich Biondi Heidenhainî method? Particularly how long has the specimen to stay in the staining solution? I have noticed, in previous, tests that is quit difficult to dye the nucleus with the methyl green contained in the solution. I am processing thin sections of mouse liver included in wax and fixed with Bouin liquid. I also wonder why the sections are crunched after cutting. There is a solution at this problem? Thanks in advance. Best Regards, Massimo
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (andromeda_tm-at-libero.it) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 3, 2006 at 04:36:47 ---------------------------------------------------------------------------
Hi everyone, would any of you give me some information about the procedure on using Aceto Carmine for nucleos staining and what can I use like contrast colour? Thank you in advance for your assistance. Best Regards, Massimo
I would like to contact anyone who has experience working with tobacco suspension cells. I am interested in general morphology information as well as preparation using standard chemical fix and high pressure freezing methods.
As this is a rather specific request, you might want to contact me off-line.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 5, 21 -- From dsherman-at-purdue.edu Fri Mar 3 08:46:44 2006 5, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k23EkhFF017338 5, 21 -- for {microscopy-at-microscopy.com} ; Fri, 3 Mar 2006 08:46:43 -0600 5, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.235]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 5, 21 -- Fri, 3 Mar 2006 09:46:41 -0500 5, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 5, 21 -- Fri, 3 Mar 2006 14:46:42 +0000 5, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 5, 21 -- Date: Fri, 03 Mar 2006 09:46:41 -0500 5, 21 -- Subject: Tobacco suspension cells 5, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 5, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 5, 21 -- Message-ID: {C02DC101.DCF0%dsherman-at-purdue.edu} 5, 21 -- Thread-Topic: Tobacco suspension cells 5, 21 -- Thread-Index: AcY+0UhghzF7x6rEEdq0gQARJN08Mg== 5, 21 -- Mime-version: 1.0 5, 21 -- Content-type: text/plain; 5, 21 -- charset="US-ASCII" 5, 21 -- Content-transfer-encoding: 7bit 5, 21 -- X-OriginalArrivalTime: 03 Mar 2006 14:46:41.0564 (UTC) FILETIME=[48B66DC0:01C63ED1] ==============================End of - Headers==============================
The following is the information regarding the funeral service for Dr. Apkarian. You can also view the announcement at http://www.chemistry.emory.edu/. Thank you.
Hong Emory EM
} We regret to announce that Dr. Robert Apkarian died in an traffic } accident on Tuesday, February 28, 2006. The Department of Chemistry } extends our deepest condolences to the Apkarian family. } } The funeral will be Monday, March 6th. } at the } Greek Orthodox Church } 2480 Clairmont Road, NE } Atlanta, GA 30329 } } 10:00 to 11:00 a.m. Viewing } 11:00 - 12:00 Service } Reception immediately following } } } All are welcome to attend. } } Please assist us by forwarding this information to any of Dr. } Apkarian's } friends who we may not have reached. } } Thank you.
==============================Original Headers============================== 7, 21 -- From hyi-at-emory.edu Fri Mar 3 09:01:29 2006 7, 21 -- Received: from pales.cc.emory.edu (pales.cc.emory.edu [170.140.8.221]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k23F1SWm021950 7, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Mar 2006 09:01:28 -0600 7, 21 -- Received: from [170.140.233.152] (localhost [127.0.0.1]) 7, 21 -- by pales.cc.emory.edu (8.13.4/8.13.4) with ESMTP id k23F1RlK014558 7, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Mar 2006 10:01:27 -0500 (EST) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v622) 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- Message-Id: {96f4f9509ca6223516484dc8bdf4b16d-at-emory.edu} 7, 21 -- Content-Type: text/plain; 7, 21 -- charset=US-ASCII; 7, 21 -- format=flowed 7, 21 -- To: Microscopy-at-microscopy.com 7, 21 -- From: Hong Yi {hyi-at-emory.edu} 7, 21 -- Subject: Fwd: [Fwd: Dr. Rob Apkarian] 7, 21 -- Date: Fri, 3 Mar 2006 10:01:26 -0500 7, 21 -- X-Mailer: Apple Mail (2.622) 7, 21 -- X-imss-version: 2.038 7, 21 -- X-imss-result: Passed 7, 21 -- X-imss-approveListMatch: *-at-emory.edu ==============================End of - Headers==============================
We are having trouble with our old LifeCell freezing machine (a slam freezer) so we would be interested in obtaining one that someone has laying around in the corner. It does not even have to be functioning as we have some clever folks here who might be able to fix ours with old parts. Please contact me with any ideas, machines or parts that might be out there somewhere.
There is a job open at Children's Hospital San Diego for a TEM/Histologist. The person will be responsible for doing the majority of EM (the EM workload varies, though it is almost exclusively renal tissue) and assisting in the Histology section when not doing EM (which happens frequently). So histology skills are an obvious plus.
If you are interested please contact me at psicurello-at-chsd.org or Dr. Eric Breisch at ebreisch-at-chsg.org. Please include a copy of your CV or resume.
Sunny San Diego awaits you!
Paula Sicurello :-)
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==============================Original Headers============================== 3, 18 -- From patpxs-at-yahoo.com Fri Mar 3 12:09:00 2006 3, 18 -- Received: from web52202.mail.yahoo.com (web52202.mail.yahoo.com [206.190.48.125]) 3, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k23I8xTd000725 3, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 3 Mar 2006 12:08:59 -0600 3, 18 -- Received: (qmail 2020 invoked by uid 60001); 3 Mar 2006 18:08:59 -0000 3, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 3, 18 -- s=s1024; d=yahoo.com; 3, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 3, 18 -- b=K/KClLe8EWmfVPKcUH9yRpeaPR7Tb2H5gmmoQQ66aLtQrbP98K5C2hRzZ9mW0ERYeDHLoh3/pxARyc4IUUb5kvJmhCmdKTW/0cT5GyjqGJKJPuDEMLi6aEmtMhiiDLnLokMFx7DAqMEzrTOifo1OJ+BaIO6zhfBivbFMN8vx7UE= ; 3, 18 -- Message-ID: {20060303180859.2018.qmail-at-web52202.mail.yahoo.com} 3, 18 -- Received: from [209.203.68.199] by web52202.mail.yahoo.com via HTTP; Fri, 03 Mar 2006 10:08:59 PST 3, 18 -- Date: Fri, 3 Mar 2006 10:08:59 -0800 (PST) 3, 18 -- From: Paula Sicurello {patpxs-at-yahoo.com} 3, 18 -- Subject: TEM/Histologist position open in San Diego 3, 18 -- To: MSA BB {microscopy-at-msa.microscopy.com} 3, 18 -- MIME-Version: 1.0 3, 18 -- Content-Type: text/plain; charset=iso-8859-1 3, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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Question: We are asking if anyone has a V. Avarlaid Autoradiographic slide coating machine they would like to unload? Parts of ours are missing and they are no longer made. Replys can be made to me at fremingt-at-fhcrc.org. Thanks.
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Email: gibi55-at-yahoo.com Name: Gino Bianchi
Organization: Universidad Central de Venezuela
Title-Subject: [Filtered] Autotechnicon wanted
Question: Looking for an Autotechnicon mono or duo in working order
I'm with Balzers, Inc., a supplier of hard coating equipment and services. We have just opened an Applications Support Center in Elgin, IL. We are in need (hopefully temporarily) of SEM service nearby.
If anyone knows of or supplies service in this area, I would appreciate this in formation.
Best regards,
Erik
C. Erik Bauer Tool Coating Specialist Balzers, Inc. Applications Support Center 1181 Jansen Farms Ct. Elgin, IL 60123 tel: 847-695-5200 ext.2001 cell: 224-730-084 fax: 847-695-4051 erik.bauer-at-balzers.com www.balzers.com
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==============================Original Headers============================== 8, 20 -- From cerikbauer-at-yahoo.com Mon Mar 6 08:31:50 2006 8, 20 -- Received: from web60421.mail.yahoo.com (web60421.mail.yahoo.com [209.73.178.149]) 8, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k26EVmCG015478 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 6 Mar 2006 08:31:48 -0600 8, 20 -- Received: (qmail 11744 invoked by uid 60001); 6 Mar 2006 14:31:48 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=Nim4mrtWgLFwHgT2ibLpYVAfN0b75xTX+I0fBLsBpxSVAD6+8Q0SOW/4vwLJh/5jJLElN/O8yYV4O2cfDQaVUa4fuDMFY6IpH9oP+Zpjv8q5jbsMhXnx5XWQFFhcqPOsI6S+0sB+nL7OxkvVxcVSOjrJEyVCY5rJbLEE9aonQlI= ; 8, 20 -- Message-ID: {20060306143148.11742.qmail-at-web60421.mail.yahoo.com} 8, 20 -- Received: from [63.144.89.98] by web60421.mail.yahoo.com via HTTP; Mon, 06 Mar 2006 06:31:48 PST 8, 20 -- Date: Mon, 6 Mar 2006 06:31:48 -0800 (PST) 8, 20 -- From: Erik Bauer {cerikbauer-at-yahoo.com} 8, 20 -- Subject: SEM Services near Chicago 8, 20 -- To: List Microscopy {Microscopy-at-MSA.Microscopy.Com} 8, 20 -- Cc: "Dennis T. Quinto" {dennis.quinto-at-balzers.com} , 8, 20 -- volker.derflinger-at-balzers.com 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
We will have a full time job opening for an entry level EM Tech. This position will support our current operation for clinical (hospital) as well as our research needs (medical school).
Thank you.
Rajesh Patel Imaging Suite Rm 024 School of Public Health 683 Hoes Lane Piscataway, NJ 08854
Just a word of caution, there are health/safety issues in dealing with inorganic fluorine compounds. Make sure you find the proper MSDS's for what you're dealing with.
Jane L. LaGoy Lab Manager/R&D Engineer Bodycote North America 155 River Street Andover, MA 01810 978-470-1620 x450 FAX: 978-475-2951 jane.lagoy-at-bodycote.com The only people to get even with are those who have helped you.
-----Original Message----- X-from: randy-nessler-at-uiowa.edu [mailto:randy-nessler-at-uiowa.edu] Sent: Tuesday, February 28, 2006 6:06 PM To: jane.lagoy-at-bodycote.com
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please copy both randy-nessler-at-uiowa.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: randy-nessler-at-uiowa.edu Name: Randy Nessler
Organization: University of Iowa
Title-Subject: [Filtered] Fluorine removal?
Question: I have been asked to post this question for a collegue. It appears that he might be getting fluorine contamination of his XPS samples when they are in a processing chamber. The chamber is lined with glass. Is there a suitable reagent to clean this chamber out with to remove any residual fluorine? "Baking" the chamber hasn't minimied the problem. Thanks, Randy
The seventh annual European ESEM Userclub meeting will take place in London on June 26th 2006, preceeding the Royal Microscopical Society's MICROSCIENCE 2006 conference.
For more information & registration, please visit:
http://www.rms.org.co.uk/events_esem/shtml
(By the way, to register for MICROSCIENCE 2006, June 27th - 29th, please visit http://www.microscience2006.org.uk/conference_registration.shtml. Note that the abstract deadline is 3rd April 2006).
Best Wishes,
Debbie.
-- Dr Debbie Stokes
Biological & Soft Systems University of Cambridge Dept of Physics Cavendish Laboratory JJ Thomson Avenue Cambridge CB3 0HE
I need to glue a little plastic ring to the end of a TEM cryoholder. Armstrong A12 was recommended, but I don't have any around. Anyone know where I can get some or if any epoxy is OK? Especially concerned about something that is good with vacuum and cryo temps.
I have some old M-bond 610 around that might do, but the directions say its shelf life is only about 9 months even unmixed. Mine is older than that, is it really no good?
Thanks
Jon -- Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 5, 21 -- From jmkrupp-at-ucsc.edu Mon Mar 6 16:44:18 2006 5, 21 -- Received: from smtp-prod-mx2.ucsc.edu (smtp-prod-mx2.ucsc.edu [128.114.125.44]) 5, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k26MiHeF006991 5, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Mar 2006 16:44:17 -0600 5, 21 -- Received: from ucsc.edu (cruzmail-fe1.ucsc.edu [128.114.125.5]) 5, 21 -- by smtp-prod-mx2.ucsc.edu (8.13.1/8.13.1) with ESMTP id k26MaAfw024370 5, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Mar 2006 14:36:11 -0800 (PST) 5, 21 -- Received: from [128.114.25.190] (account jmkrupp-at-ucsc.edu [128.114.25.190] verified) 5, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 5, 21 -- with ESMTPA id 48085986 for Microscopy-at-Microscopy.Com; Mon, 06 Mar 2006 14:36:05 -0800 5, 21 -- Mime-Version: 1.0 5, 21 -- Message-Id: {p06230903c032684ff6f1-at-[128.114.25.190]} 5, 21 -- Date: Mon, 6 Mar 2006 14:36:04 -0800 5, 21 -- To: Microscopy-at-microscopy.com 5, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 5, 21 -- Subject: Armstrong A12 5, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 5, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 5, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 5, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
As a brand new Hitachi user (S-3400N II), is was somewhat amazed to find that there is no dedicated Hitachi user listserver.
I also run a Cameca SX51 electron probe and with several other users in 1994 initiated the sx50-users listserver that has been infinitely useful for the past 12 years. And a JEOL epma list started a couple of years ago.
So why no listserver for Hitachi SEM users? It would seem that it would be quite useful for both new and experienced users, and helpful for sorting out a variety of issues and problems specific to users of these flavor instruments.
If you are interested, please contact me off line (off the list), direct to johnf-at-geology.wisc.edu
John Fournelle -- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
==============================Original Headers============================== 7, 24 -- From johnf-at-geology.wisc.edu Mon Mar 6 18:02:33 2006 7, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 7, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2702XOU026881 7, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 6 Mar 2006 18:02:33 -0600 7, 24 -- Received: from localhost (localhost [127.0.0.1]) 7, 24 -- by localhost (Postfix) with ESMTP id 0663620D1A 7, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 6 Mar 2006 18:02:33 -0600 (CST) 7, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 7, 24 -- by localhost (ice [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 7, 24 -- id 05536-09 for {Microscopy-at-Microscopy.Com} ; 7, 24 -- Mon, 6 Mar 2006 18:02:30 -0600 (CST) 7, 24 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 7, 24 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 7, 24 -- (No client certificate requested) 7, 24 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 7C5C320D0C 7, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 6 Mar 2006 18:02:30 -0600 (CST) 7, 24 -- Mime-Version: 1.0 7, 24 -- Message-Id: {p06230906c0327be78b16-at-[144.92.206.57]} 7, 24 -- Date: Mon, 6 Mar 2006 17:59:44 -0600 7, 24 -- To: Microscopy List_nestors {Microscopy-at-Microscopy.Com} 7, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 7, 24 -- Subject: Hitachi SEM Users? 7, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 24 -- X-Virus-Scanned: by amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
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Email: Dorothy.Howard-at-noaa.gov Name: Dorothy Howard
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Email: fulton.2-at-osu.edu Name: Dave Fulton
Organization: OARDC/MCIC/Ohio State University
Title-Subject: [Filtered] Microwave tissue processing for TEM
Question: Hello fellow Listers; Does anyone out there have a Microwave Research and Applications, Inc. , model BP-111-RS laboratory microwave oven, which they are using to process tissue samples for TEM? We have recently purchased one and have made several attempts to use it to process plant tissue (maize and tobacco) for TEM without success. If you have successful protocols that you have used with this oven and would be willing to share them, it would be greatly appreciated. You may reply directly to me if you wish. Thanks in advance for your time and trouble.
Dorothy Howard wrote: ========================================== Question: Could you tell me where in Maryland or Delaware you can contract for EM services? ==========================================
Would "just over the border in Pennsylvania" be considered? Structure Probe, Inc. has been offering both SEM and TEM services since 1970 as a laboratory service in West Chester, PA. Our contact information if given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 14, 25 -- From cgarber-at-2spi.com Mon Mar 6 22:32:44 2006 14, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 14, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k274WgUX017564 14, 25 -- for {microscopy-at-msa.microscopy.com} ; Mon, 6 Mar 2006 22:32:43 -0600 14, 25 -- Received: from ibm1x23g2abfyg (217-118-122-167.client.stsn.net [217.118.122.167]) 14, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k274WdIY015703; 14, 25 -- Mon, 6 Mar 2006 23:32:41 -0500 14, 25 -- X-IDV-FirstRcvd: 217-118-122-167.client.stsn.net [217.118.122.167] 14, 25 -- X-IDV-HELO: ibm1x23g2abfyg 14, 25 -- Message-ID: {02b101c641a0$298ac190$4b3f680a-at-ibm1x23g2abfyg} 14, 25 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 14, 25 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 14, 25 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 14, 25 -- Cc: {Dorothy.Howard-at-noaa.gov} 14, 25 -- Subject: TEM and SEM contract services in DE and MD 14, 25 -- Date: Mon, 6 Mar 2006 23:32:35 -0500 14, 25 -- MIME-Version: 1.0 14, 25 -- Content-Type: text/plain; 14, 25 -- charset="Windows-1252" 14, 25 -- X-Priority: 3 14, 25 -- X-MSMail-Priority: Normal 14, 25 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 14, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 14, 25 -- Content-Transfer-Encoding: 8bit 14, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k274WgUX017564 ==============================End of - Headers==============================
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Email: g.posthuma-at-lab.azu.nl Name: George Posthuma
Organization: Dept of Cell Biology, Utrecht, The netherlands
Title-Subject: [Filtered] workshop on Cryomethods, Ultracryotomy and Immunolabeling
Question: dear all,
In collaboration with Leica we will organize a workshop on Cryomethods, Ultracryotomy and Immunolabeling in Utrecht, The Netherlands. If you are interested, please have a look at: http://www.cmc-utrecht.nl/education/indexeducation.htm. The maximum of participants is 16, of which already 13 are booked.
This is for all you cryo experts out there. I was contacted by a researcher who wants to look at ice crystals in thin sections of ice cream in the TEM. Anyone out there who has the capability to do this sort of cryo work? The researcher is located at the University of Nebraska-Lincoln, Lincoln, NE. We do not have the cryo capability here in Nebraska. So I would like to find out who out there is closest and then we'll see if we can find a way to get the ice cream to you without it melting. This individual is not an EM person so I offerred to try to gather some information for him to see what might be available in the way of help. If anyone out there can help, please contact me and I'll pass the information along. Thanks
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center Omaha, NE 68198-6395 e-mail: tbargar-at-unmc.edu phone: 402-559-7347 fax: 402-559-3400
==============================Original Headers============================== 5, 20 -- From tbargar-at-unmc.edu Tue Mar 7 09:19:18 2006 5, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27FJIJs015410 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:19:18 -0600 5, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 5, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id A2F00A00AF 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:15 -0600 (CST) 5, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 5, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 30DAE398085 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:14 -0600 (CST) 5, 20 -- Subject: Need cryoultramicrotomy of ice cream 5, 20 -- To: Microscopy-at-MSA.Microscopy.com 5, 20 -- X-Mailer: Lotus Notes Release 6.5.1 January 21, 2004 5, 20 -- Message-ID: {OFA994B2B5.45E530B5-ON8625712A.0052F932-8625712A.005428D0-at-unmc.edu} 5, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 5, 20 -- Date: Tue, 7 Mar 2006 09:19:14 -0600 5, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR(Release 6.5.5 5, 20 -- HF62|January 19, 2006) at 03/07/2006 09:19:16 AM 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Um ... any particular reason for the thin sections? CryoSEM of ice cream has been done to examine ice crystals, lipid droplets, and air spaces. I'd think this would be a more useful (and much less stressful) technique than is cryoultrasectioning and cryo TEM and opening the crystals don't change in the process, then watching them change in the beam ... Lincoln Lim did this with the cryo stage on the Hitachi S-900 at UW-Madison, using low kV (~1.5, if I remember right), the researcher might want to look for his publications. Are there any cryoSEM labs around Lincoln?
Phil
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==============================Original Headers============================== 3, 23 -- From oshel1pe-at-cmich.edu Tue Mar 7 10:22:22 2006 3, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27GMMGG025898 3, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 10:22:22 -0600 3, 23 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 3, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k27H0r4l012285; 3, 23 -- Tue, 7 Mar 2006 12:00:53 -0500 3, 23 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 3, 23 -- Tue, 7 Mar 2006 11:20:51 -0500 3, 23 -- Mime-Version: 1.0 3, 23 -- Message-Id: {f06230902c03362a1cb3f-at-[141.209.160.132]} 3, 23 -- In-Reply-To: {200603071604.k27G4guS024098-at-ns.microscopy.com} 3, 23 -- References: {200603071604.k27G4guS024098-at-ns.microscopy.com} 3, 23 -- Date: Tue, 7 Mar 2006 11:22:17 -0500 3, 23 -- To: tbargar-at-unmc.edu 3, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 23 -- Subject: Re: [Microscopy] Need cryoultramicrotomy of ice cream 3, 23 -- Cc: Microscopy-at-microscopy.com 3, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 23 -- X-OriginalArrivalTime: 07 Mar 2006 16:20:51.0689 (UTC) FILETIME=[1A1A1590:01C64203] 3, 23 -- X-CanItPRO-Stream: default 3, 23 -- X-Spam-Score: -4 () L_EXCH_MF 3, 23 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Check out the "foods under the microscope" web page: http://www.magma.ca/~scimat/ This is a good resource on a variety of microscopic techniques on dairy products. They discuss the microscopy of yogurt and cheeses, as well as dried milk products. The bibliography is extensive.
Karl -----Original Message----- X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu] Sent: Tuesday, March 07, 2006 10:39 AM To: Karl Hagglund
This is for all you cryo experts out there. I was contacted by a researcher who wants to look at ice crystals in thin sections of ice cream in the TEM. Anyone out there who has the capability to do this sort of cryo work? The researcher is located at the University of Nebraska-Lincoln, Lincoln, NE. We do not have the cryo capability here in Nebraska. So I would like to find out who out there is closest and then we'll see if we can find a way to get the ice cream to you without it melting. This individual is not an EM person so I offerred to try to gather some information for him to see what might be available in the way of help. If anyone out there can help, please contact me and I'll pass the information along. Thanks
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center Omaha, NE 68198-6395 e-mail: tbargar-at-unmc.edu phone: 402-559-7347 fax: 402-559-3400
==============================Original Headers============================== 5, 20 -- From tbargar-at-unmc.edu Tue Mar 7 09:19:18 2006 5, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27FJIJs015410 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:19:18 -0600 5, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 5, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id A2F00A00AF 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:15 -0600 (CST) 5, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 5, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 30DAE398085 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Tue, 7 Mar 2006 09:36:14 -0600 (CST) 5, 20 -- Subject: Need cryoultramicrotomy of ice cream 5, 20 -- To: Microscopy-at-MSA.Microscopy.com 5, 20 -- X-Mailer: Lotus Notes Release 6.5.1 January 21, 2004 5, 20 -- Message-ID: {OFA994B2B5.45E530B5-ON8625712A.0052F932-8625712A.005428D0-at-unmc.edu} 5, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 5, 20 -- Date: Tue, 7 Mar 2006 09:19:14 -0600 5, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR(Release 6.5.5 5, 20 -- HF62|January 19, 2006) at 03/07/2006 09:19:16 AM 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 13, 24 -- From hagglundk1-at-nku.edu Tue Mar 7 13:46:27 2006 13, 24 -- Received: from mailfe2.nku.edu (exchange.nku.edu [192.122.237.68]) 13, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27JkRpE006150 13, 24 -- for {microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 13:46:27 -0600 13, 24 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 13, 24 -- Tue, 7 Mar 2006 14:46:26 -0500 13, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 13, 24 -- Content-class: urn:content-classes:message 13, 24 -- MIME-Version: 1.0 13, 24 -- Content-Type: text/plain; 13, 24 -- charset="us-ascii" 13, 24 -- Subject: RE: [Microscopy] Need cryoultramicrotomy of ice cream 13, 24 -- Date: Tue, 7 Mar 2006 14:46:26 -0500 13, 24 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F358691-at-mailfac1.hh.nku.edu} 13, 24 -- X-MS-Has-Attach: 13, 24 -- X-MS-TNEF-Correlator: 13, 24 -- Thread-Topic: [Microscopy] Need cryoultramicrotomy of ice cream 13, 24 -- Thread-Index: AcZCALuBxcuDWRkORCi4uvSXIKSRKwAHiweQ 13, 24 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 13, 24 -- To: {tbargar-at-unmc.edu} 13, 24 -- Cc: {microscopy-at-microscopy.com} 13, 24 -- X-OriginalArrivalTime: 07 Mar 2006 19:46:26.0709 (UTC) FILETIME=[D258C450:01C6421F] 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k27JkRpE006150 ==============================End of - Headers==============================
George also comes to the International Cryo-EM course at the University of British Columbia, Vancouver, British Columbia, Canada hosted by Kent McDonald, U Berkeley and Elaine Humphrey, UBC
This is a 10-day course, June 6-15, 2006, covering Cryo-TEM, Cryo-SEM, high pressure freezing, Cryo Ultramicrotomy, Tokuyasu method and immunolabelling.
This year we have new instruments from Leica and Baltec. follow the links from http://www.emlab.ubc.ca
Elaine
} } Email: g.posthuma-at-lab.azu.nl } Name: George Posthuma } } Organization: Dept of Cell Biology, Utrecht, The netherlands } } Title-Subject: [Filtered] workshop on Cryomethods, Ultracryotomy and } Immunolabeling } } Question: dear all, } } In collaboration with Leica we will organize a workshop on } Cryomethods, Ultracryotomy and Immunolabeling in Utrecht, The } Netherlands. If you are interested, please have a look at: } http://www.cmc-utrecht.nl/education/indexeducation.htm. The maximum } of participants is 16, of which already 13 are booked. } } Yours sincerely } } George Posthuma }
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada (2003-2005) University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
==============================Original Headers============================== 7, 30 -- From ech-at-interchange.ubc.ca Tue Mar 7 14:37:18 2006 7, 30 -- Received: from mta3.mail-relay.ubc.ca (mta3.mail-relay.ubc.ca [137.82.45.6]) 7, 30 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27KbGC1015821 7, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Mar 2006 14:37:16 -0600 7, 30 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 7, 30 -- by mta3.mail-relay.ubc.ca (8.12.11/8.12.11) with ESMTP id k27KbDs3025904 7, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Mar 2006 12:37:14 -0800 (PST) 7, 30 -- (envelope-from ech-at-interchange.ubc.ca) 7, 30 -- Received: from [137.82.85.193] (echpowerbook.emlab.ubc.ca [137.82.85.193]) 7, 30 -- by smtp.interchange.ubc.ca 7, 30 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 7, 30 -- with ESMTPA id {0IVR00FP9YLJ63-at-smtp.interchange.ubc.ca} for 7, 30 -- microscopy-at-msa.microscopy.com; Tue, 07 Mar 2006 12:36:55 -0800 (PST) 7, 30 -- Date: Tue, 07 Mar 2006 12:36:54 -0800 7, 30 -- From: Elaine Humphrey {ech-at-interchange.ubc.ca} 7, 30 -- Subject: Re: [Microscopy] viaWWW: workshop on Cryomethods, 7, 30 -- Ultracryotomy and Immunolabeling 7, 30 -- In-reply-to: {200603071425.k27EPhpY007742-at-ns.microscopy.com} 7, 30 -- X-Sender: ech-at-mail.interchange.ubc.ca 7, 30 -- To: microscopy-at-msa.microscopy.com 7, 30 -- Cc: g.posthuma-at-lab.azu.nl 7, 30 -- Message-id: {a06100501c03376ebe414-at-[137.82.85.193]} 7, 30 -- MIME-version: 1.0 7, 30 -- Content-type: text/plain; format=flowed; charset=us-ascii 7, 30 -- References: {200603071425.k27EPhpY007742-at-ns.microscopy.com} 7, 30 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.03.07.115105 7, 30 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 7, 30 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0 7, 30 -- X-Spam-Level: 7, 30 -- X-Spam-Flag: No ==============================End of - Headers==============================
THE TEXAS SOCIETY FOR MICROSCOPY invites you to our Spring 2006 meeting on April 20-22, 2006 at Alcon Research Labs, 6201 South Freeway, Ft. Worth, TX 76134 2ND CALL FOR PAPERS
All registration forms and lodging details are available on our web site: http://www.texasmicroscopy.org/
ABSTRACTS MUST BE RECEIVED BY: March 20, 2006 Advanced Registration Deadline: April 7, 2006. Advanced registration is strongly suggested to afford TSM an accurate participant count for event organization AND to comply with Alcon security requirements. Thursday workshops and Friday sessions are being held at Alcon Research.
**Workshops— Thursday, April 20, 2006
“Microwave Processing: Factors the Influence Results” Sponsored by Ted Pella, Inc. Speaker: Rick Giberson, Sr. Applications Engineer
“ESEM: not just for Biology Anymore” Sponsored by FEI Company Speaker: Daniel Phifer, Sr. Applications Engineer
**Guest Speaker — Friday, April 21, 2006
“Materials Science in Museums” Dr. Pamela Vandiver, Professor of Materials Science and Engineering and Archeology, Co-Director of Program in Heritage Conservation Science at the University of Arizona, and former Senior Research Scientist at the Smithsonian Institution’s Center for Materials Research and Education.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 12, 21 -- From r-holdford-at-ti.com Tue Mar 7 15:43:20 2006 12, 21 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 12, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k27LhJZS026508 12, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Mar 2006 15:43:19 -0600 12, 21 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 12, 21 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k27LhAjT009990 12, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Mar 2006 15:43:16 -0600 (CST) 12, 21 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 12, 21 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k27LhANd018028 12, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Mar 2006 15:43:10 -0600 (CST) 12, 21 -- Message-ID: {440DFE6E.2010101-at-ti.com} 12, 21 -- Date: Tue, 07 Mar 2006 15:43:10 -0600 12, 21 -- From: Becky Holdford {r-holdford-at-ti.com} 12, 21 -- Organization: SC Packaging Development -- FA Development 12, 21 -- User-Agent: Mozilla Thunderbird 0.8 (Windows/20040913) 12, 21 -- X-Accept-Language: en-us, en 12, 21 -- MIME-Version: 1.0 12, 21 -- To: MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 12, 21 -- Subject: 2nd call for papers for Spring meeting of the Texas Society for Microscopy 12, 21 -- Content-Type: text/plain; charset=windows-1252; format=flowed 12, 21 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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 camiller-at-anatomy.iupui.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: camiller-at-anatomy.iupui.edu Name: Caroline A. Miller
Organization: Indiana University, Indiana Microscopy Society
Title-Subject: [Filtered] Our upcoming Spring Meeting, March 20th, 2006
Question: The Spring Meeting of the Indiana Microscopy Society will be Monday, March 20 at the University of Notre Dame, South Bend, IN 8 AM until 3:15 PM in the McKenna Hall Conference Center
Guest Speakers: Daphna Yaniv, PhD, Electron Microscopy Sciences/Quantomix ěEM of Fully Hydrated Samplesî
Eva Chi, PhD, University of Chicago ěRole of Cell Membrane in the Pathogenisis of Alzheimerís Diseaseî
Alex Kandel, PhD, University of Notre Dame ěStructure and Dynamics of Organic Molecules on Surfaces, One Molecule at a Timeî
There will be a best Micrograph and Student Poster Competition A Tour of the Notre Dame Campus will be offered at 3:15 Registration is $10 for members, $20 for non-members Students are free with membership
Breakfast and Lunch Provided For registration and more information go to: indianamicroscopy.org
A very good day to all of you! I apologise that this might be out of place. I am doing some UV-Vis absorption test on my powders in different solvent. I am intending to use carbon tetrachloride or chloroform with a quartz cuvette. However, I am worry that the solvent might damage the lining/joining part of the quartz cuvette. I was informed that strong acid will damage it but I am not too sure CCl4 or chloroform.
I would kindly seek advise from you regarding this. Thank you so much in advance first!!
Have a nice day ahead!
Cheers, YY School of Materials Science and Engineering Nanyang Technological University Singapore
__________________________________ Do you Yahoo!? Yahoo! Movies - Search movie info and celeb profiles and photos. http://sg.movies.yahoo.com/
==============================Original Headers============================== 7, 18 -- From one_twinklestar-at-yahoo.com.sg Tue Mar 7 18:47:47 2006 7, 18 -- Received: from web50006.mail.yahoo.com (web50006.mail.yahoo.com [206.190.38.21]) 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k280lkHm011734 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 18:47:46 -0600 7, 18 -- Received: (qmail 4555 invoked by uid 60001); 8 Mar 2006 00:47:45 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com.sg; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=PhDz60Edu3BvwxWyY+MwzR9o7q7TrMoXIWIFKaqnRTmP64hZzF0dBNFXJS4ocu0rU9Rb6skVASz5goyEUUIxemAXOxr+lHrMk03GGuAdvKACiCU5CuIf0ApYQhlCpFf6eBTqvLxKUCLqp+BDQWX15qow0Vb3OzoTLwoRUH4hnp4= ; 7, 18 -- Message-ID: {20060308004745.4553.qmail-at-web50006.mail.yahoo.com} 7, 18 -- Received: from [202.21.158.12] by web50006.mail.yahoo.com via HTTP; Wed, 08 Mar 2006 08:47:45 CST 7, 18 -- Date: Wed, 8 Mar 2006 08:47:45 +0800 (CST) 7, 18 -- From: Tay Yee Yan {one_twinklestar-at-yahoo.com.sg} 7, 18 -- Subject: Regarding Carbon TetraChloride or Chloroform with Quart Cuvette 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; charset=iso-8859-1 7, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
It's a few decades since I've used such things, but I'm fairly sure they are one-piece, ie just quartz, with no liners or joints. I doubt that strong acid, apart from HF, of course, will damage them either. I used to routinely clean them with the so-called chromic acid (sodium dichromate dissolved in concentrated sulfuric acid) with no problems at all.
cheers
rtch
On 7 Mar 2006 at 19:02, one_twinklestar-at-yahoo.com.sg wrote:
} } } } ---------------------------------------------------------------------------- } 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: } } A very good day to all of you! I apologise that this } might be out of place. I am doing some UV-Vis } absorption test on my powders in different solvent. I } am intending to use carbon tetrachloride or chloroform } with a quartz cuvette. However, I am worry that the } solvent might damage the lining/joining part of the } quartz cuvette. I was informed that strong acid will } damage it but I am not too sure CCl4 or chloroform. } } I would kindly seek advise from you regarding this. } Thank you so much in advance first!! } } Have a nice day ahead! } } Cheers, } YY } School of Materials Science and Engineering } Nanyang Technological University } Singapore } }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 10, 27 -- From r.sims-at-auckland.ac.nz Tue Mar 7 19:41:23 2006 10, 27 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) 10, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k281fLnO024295 10, 27 -- for {microscopy-at-microscopy.com} ; Tue, 7 Mar 2006 19:41:21 -0600 10, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 10, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 04F693435F; 10, 27 -- Wed, 8 Mar 2006 14:41:20 +1300 (NZDT) 10, 27 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) 10, 27 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 27 -- with ESMTP id 10821-05; Wed, 8 Mar 2006 14:41:19 +1300 (NZDT) 10, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id DAA7234118; 10, 27 -- Wed, 8 Mar 2006 14:41:19 +1300 (NZDT) 10, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 27 -- To: one_twinklestar-at-yahoo.com.sg 10, 27 -- Date: Wed, 08 Mar 2006 14:44:43 +1300 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] Regarding Carbon TetraChloride or Chloroform with Quart Cuvette 10, 27 -- Cc: microscopy-at-microscopy.com 10, 27 -- Message-ID: {440EEDDB.5137.1427CB3-at-localhost} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200603080102.k2812K8Q015619-at-ns.microscopy.com} 10, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 10, 27 -- Content-type: text/plain; charset=US-ASCII 10, 27 -- Content-transfer-encoding: 7BIT 10, 27 -- Content-description: Mail message body 10, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (nomy_nay-at-hotmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, March 7, 2006 at 22:09:53 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both nomy_nay-at-hotmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: In electron microscopy, the higher the voltage the greater the penetrating abilityof the electron beam, but the trade is a reduction of what?
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Email: sue.tyler-at-noaa.gov Name: Sue Tyler
Organization: NOAA
Title-Subject: [Filtered] Pafaffin block storage
Question: Our facility is in the process of building a new paraffin block storage building. The question is, are there specific temperature ranges to maintain optimal block preservation?
Our outside temps. in Maryland are as low as 0 degrees C in the winter.
I have searched the histonet archives but did not come up with any specific temp. Perhaps someone could offer some suggestions. Thanks
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] Disposing of Dark Room Chemicals
Question: Lately we have been asked to review our procedures for disposing of chemicals. I was wondering if Kodak has any official recommendations for disposing of D-19 developing and Kodak fixer for TEM negatives.
Higher voltage may 1) Damage your samples easily. For Al foil, you can easily see the beam damage at 300kv 2) Cause multibeam effect when you use the diffraction contrast techniques. Short wavelength means flat Ewards sphere, or more beams are excited.
OF course, cost and maintenance are also problems.
---- Original message ---- } Date: Wed, 8 Mar 2006 08:59:51 -0600 } From: nomy_nay-at-hotmail.com } Subject: [Microscopy] AskAMicroscopist: higher voltage decreases what? } To: clei-at-uiuc.edu } } } } } ------------------------------------------------------------- --------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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Mark, You probably need to check your regional/institutional requirements. Here in NYC we are required to use a silver capture system to filter our photographic fix solutions. If we do that, then the D-19 can do down the drain with a lot of water. If we don't use a silver capture system, then we must collect all solutions (developer, stop and fix) and have our Environmental & Life Safety officers take it all away for disposal. If you have a low volume, you can get a simple gravity filtration system that you just pour the fix through when its exhausted. If you have an automated processor, there are traps that can be connected in series with the rest of the system to capture the silver out of the fix. We have both types of systems here, and they work well. The outside contractor comes through periodically to monitor them and change the active capture filter when needed. Its actually pretty painless, and not too expensive. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 21 -- From lcgould-at-med.cornell.edu Wed Mar 8 09:30:22 2006 1, 21 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28FUKbZ031038 1, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:30:20 -0600 1, 21 -- Received: from mpx2.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 21 -- by smtp-gw2.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k28FUHRL028004 1, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:30:18 -0500 (EST) 1, 21 -- Received: from [140.251.48.23] by mpx2.med.cornell.edu 1, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 21 -- with ESMTP id {0IVT00BZDF2HX5A0-at-mpx2.med.cornell.edu} for 1, 21 -- microscopy-at-microscopy.com; Wed, 08 Mar 2006 10:30:17 -0500 (EST) 1, 21 -- Date: Wed, 08 Mar 2006 10:26:02 -0500 1, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 21 -- Subject: Re: [Microscopy] viaWWW: Disposing of Dark Room Chemicals 1, 21 -- In-reply-to: {200603081520.k28FKv8w028026-at-ns.microscopy.com} 1, 21 -- To: twigg-at-estd.nrl.navy.mil, Microscopy Listserver {microscopy-at-microscopy.com} 1, 21 -- Message-id: {p06200709c034a61c8ce8-at-[140.251.48.23]} 1, 21 -- MIME-version: 1.0 1, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 21 -- References: {200603081520.k28FKv8w028026-at-ns.microscopy.com} 1, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.03.08.065107 ==============================End of - Headers==============================
I have some basic very technical questions. I want to prepare MCF-7, HepG2 and Caco-2 cells for TEM observation. My protocol involves the following: - grow cells on 6W multiwell plates up to 70-80% confluency - Wash cells 1x with cold PBS - Add 1 ml cold PBS and detach cells with a cell scraper - Collect the cells in a 1.5 ml eppendorf tube, rince the well 1x with 500µl PBS and merge the volumes ----| total 1.5 ml - Centrifuge at 4°C for 5 min at 1500 RPM - Pipet out carefully the supernatant and carefully add cold Karnovsky fixative - ….
When I follow this protocol with these cells, only the HepG2 give a nice pellet, the other give a too small pellet, Please could you share with me your opinion about this protocol? - Should I forget 6W wells and grow cells on normal petri dishes? I would like to avoid that because I plan to prepare 24 conditions in parallel and working with 24 petri dishes will be a pain. - Is the centrifugation sufficient? Should I increase the centrifugation speed? - Do you have a working protocol for the preparation of these cells for morphological observation? Actually growing these cells in a way that they arrive at the same confluency at the same time is a real challenge since their growth rate are very different. This means that at least one cell line won’t be at optimal confluency at the time of the experiment. I know it’s a question of experience, but I don’t want to want 1 month before starting this experiment :-)
Another question: in parallel to this protocol, I am trying to develop a protocol for the embedding of cell monolayers. The first attempt was not too bad, the embedding basically worked, but when I try to detach the Epon resin from the bottom of the petri dish (I cutted a square with a saw), the surface of the resin is not flat. In addition I don’t know where to cut my pyramid since I don’t see where the cells are on the resin surface. Any clue?
Thank you in advance,
Stephane
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==============================Original Headers============================== 4, 18 -- From nizets2-at-yahoo.com Wed Mar 8 09:39:20 2006 4, 18 -- Received: from web37408.mail.mud.yahoo.com (web37408.mail.mud.yahoo.com [209.191.87.61]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28FdJEq001920 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:39:20 -0600 4, 18 -- Received: (qmail 58964 invoked by uid 60001); 8 Mar 2006 15:39:19 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.com; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 4, 18 -- b=lXojiS18reQ7VfhwrGChK+qQJQofBqFpgp/cSONNcQy2bC84v0/AoWGPKo1xcDx/gMrYHPQWxBDJEWLm3VNKtKpDE+bdVglPv2ioU2n9JFt+kEOxwZOc8SZsqyApi8Kww4Xs9H10q48DO/xeyLlUc9Qk8HMsJ3x6uD+DRefo2+o= ; 4, 18 -- Message-ID: {20060308153919.58962.qmail-at-web37408.mail.mud.yahoo.com} 4, 18 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 07:39:19 PST 4, 18 -- Date: Wed, 8 Mar 2006 07:39:19 -0800 (PST) 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 18 -- Subject: protocol to prepare cells in culture for TEM 4, 18 -- To: microscopy-at-microscopy.com 4, 18 -- MIME-Version: 1.0 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 4, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Stephane, I think my protocol will help you with both of your problems. You certainly can continue to use the 6-well plates. I get samples like that quite frequently. Your fixation and dehydrations can stay the same. For years, I have used a hybrid Epon-analog resin to embed in culture dishes. I use a standard Epon formula but use the followoing compnents:
LX-112 and DMP-30 from Ladd Research Industries DDSA and NMA from Electron Microscopy Sciences
I know it seems weird, but years ago I tried all sorts things, from the "straight" formulations from each vendor to a bunch of mixtures. This one has never reacted with the plastic of the dishes.
Here is how I do the actual embedding of the cell monolayers in the dishes:
After the last 100% ethanol, I remove the alcohol and cover the bottom of the well with a layer of the resin mixture that is about 2 mm deep. I then insert embedding tubes that I've made by cutting the pyramidal bottoms off of BEEM capsules (just slice them with a fresh razor blade and be sure to insert them so that the manufactured end rather than the cut one is sitting against the dish). After I insert labels into the tubes, I put these into the oven at 60 deg. overnight. In the morning, I fill JUST the embedding tubes and return everything to the oven again to finish polymerizing. When the resin is cured, you can grab the tubes with pair of needle-nosed pliers and snap them out. Sometimes a bit of the bottom of the dish comes away with the block, but often you get a very smooth block face. If some of the dish comes up, it is easy to see under a dissecting 'scope and it comes away easily when you trim you block face. I often cut away part of the block face either to keep in reserve or to re-embed to get cross sections, then trim the rest into a narrow rectangle. When you section the resulting block en face, start at 0.25 micrometers (no thicks!), and pick up and Tol. Blue the sections as you get them. You should be able to get smooth thins within a micron or so. I usually trim a very long rectangle and then start to section in such a way that I am a few degrees off of being perfectly en face from top to bottom so that I first get sections from one edge of the rectangle and then have a lot of "acreage" to work through if I need more sections later on.
Disclaimer: I have no financial interest in either Ladd or EMS...I'm just a happy customer who believes in using what works.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 7, 21 -- From lcgould-at-med.cornell.edu Wed Mar 8 10:39:26 2006 7, 21 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28GdNd1023966 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:39:25 -0600 7, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 7, 21 -- by smtp-gw1.med.cornell.edu (Switch-3.1.7/Switch-3.1.7) with ESMTP id k28GdKbq006737 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 11:39:20 -0500 (EST) 7, 21 -- Received: from [140.251.48.23] by mpx1.med.cornell.edu 7, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 7, 21 -- with ESMTP id {0IVT00D2DI9KVU50-at-mpx1.med.cornell.edu} for 7, 21 -- microscopy-at-microscopy.com; Wed, 08 Mar 2006 11:39:20 -0500 (EST) 7, 21 -- Date: Wed, 08 Mar 2006 11:35:05 -0500 7, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 7, 21 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 7, 21 -- In-reply-to: {200603081559.k28Fxc9f009527-at-ns.microscopy.com} 7, 21 -- To: nizets2-at-yahoo.com, Microscopy Listserver {microscopy-at-microscopy.com} 7, 21 -- Message-id: {p06200701c034b2185c05-at-[140.251.48.23]} 7, 21 -- MIME-version: 1.0 7, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 7, 21 -- References: {200603081559.k28Fxc9f009527-at-ns.microscopy.com} 7, 21 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.03.08.081106 ==============================End of - Headers==============================
I think that if you scrape the cells before fixation you will have a bunch of ripped up cells that have spilled their guts all over the place. We do not scrape until after osmium. If the pellet is very small I do not resuspend it, but I do centrifuge between steps. I have worked with many an invisible pellet.
nizets2-at-yahoo.com wrote:
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Tempting to say simply that the trade is a reduction of contrast. That answer would apply if you were asking about the trade of switching from say 40KV to 100KV on the same microscope.
You can compensate for this by using a smaller objective aperture.
If you are comparing a standard 40 to 100 KV microscope with a high voltage mocroscope then the answer is less straightforward and it is not necessarily so that you have a serious contrast reduction. Perhaps an electron microscope manufacturer will see this question and give you more specific answers.
Best wishes,
Ted Dunn The EMscope Company Ltd. --- nomy_nay-at-hotmail.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question was submitted to Ask-A-Microscopist by } (nomy_nay-at-hotmail.com) } from } http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } on Tuesday, March 7, 2006 at 22:09:53 } Remember to consider the Grade/Age of the student } when considering the Question } --------------------------------------------------------------------------- } Please reply to both nomy_nay-at-hotmail.com as well } as to the Microscopy Listserver } --------------------------------------------------------------------------- } } Email: nomy_nay-at-hotmail.com } Name: Naomi Piyaratna } } Organization: Wollongong University, Australia } } Education: Undergraduate College } } Location: Wollongong, NSW, AUSTRALIA } } Title: Electron Miscroscopy. } } Question: In electron microscopy, the higher the } voltage the greater the penetrating abilityof the } electron beam, but the trade is a reduction of what? } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 8, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Mar } 8 08:40:26 2006 } 8, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 8, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k28EeP1i016821 } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 } Mar 2006 08:40:26 -0600 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } (Unverified) } 8, 12 -- Message-Id: } {p0611040bc0349d3016f2-at-[206.69.208.22]} } 8, 12 -- Date: Wed, 8 Mar 2006 08:40:25 -0600 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: nomy_nay-at-hotmail.com (by way of } Ask-A-Microscopist) } 8, 12 -- Subject: AskAMicroscopist: higher voltage } decreases what? } 8, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 10, 20 -- From drteddunne-at-yahoo.com Wed Mar 8 10:47:40 2006 10, 20 -- Received: from web33401.mail.mud.yahoo.com (web33401.mail.mud.yahoo.com [68.142.206.133]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28Glche027028 10, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:47:39 -0600 10, 20 -- Received: (qmail 95631 invoked by uid 60001); 8 Mar 2006 16:47:38 -0000 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 20 -- s=s1024; d=yahoo.com; 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 10, 20 -- b=xpdlSSAAs0maYw04oUmPXHuqrXZmk07hPoZ8sv26sWh9auopxV0y/Sk8pC2kl0jDWpfW2TuOMk+g98SFzYOPojX2MrElhArfxv/3WaIY53wt88EQsbbUfuabYnRZpoWvdipk4Hgzqjz6poJVOSsr+FJE9qaEYUtJotg0YNB7sP8= ; 10, 20 -- Message-ID: {20060308164738.95629.qmail-at-web33401.mail.mud.yahoo.com} 10, 20 -- Received: from [202.47.247.156] by web33401.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 08:47:38 PST 10, 20 -- Date: Wed, 8 Mar 2006 08:47:38 -0800 (PST) 10, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: higher voltage decreases what? 10, 20 -- To: nomy_nay-at-hotmail.com 10, 20 -- Cc: microscopy-at-microscopy.com 10, 20 -- In-Reply-To: {200603081542.k28FgiZ6003132-at-ns.microscopy.com} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 10, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
A higher voltage will also reduce amplitude contrast (see below) because the nuclei of the specimen atoms will scatter higher energy electrons less than lower energy electrons. It's quite common for biologists to use a 60kv electron beam routinely to enhance contrast for instance whereas other users may favour 80kv or more for the brighter higher resolution image.
So increasing the voltage seems to produce the same effect as using a larger objective aperture (which will also reduce contrast).
NB as this is an off-list question, I think I should clarify that there are three main types of contrast seen in the transmission electron microscope (unless someone wants to add some more) - amplitude, phase and diffraction contrasts. Amplitude contrast is particularly important up to about 50,000x magnification and heavier elements in the sample with greater nuclear mass will appear darker than lighter elements because of their ability to scatter electrons out of the electron beam.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: clei-at-uiuc.edu
On Mar 8, 2006, at 6:41 AM, nomy_nay-at-hotmail.com wrote:
} Question: In electron microscopy, the higher the voltage the greater } the penetrating abilityof the electron beam, but the trade is a } reduction of what? } Dear Naomi, Contrast. The scattering cross section decreases as electron energy increases up to about 800-1000 kV (depending on the atomic number of the material). This means that for a specific scattered fraction of incident electrons, the allowed thickness will be greater at higher energy; however, since for a given thickness the scattered fraction is smaller, the difference between what happens when the beam strikes your specimen and when it passes through a hole will be less, so there is less contrast. There is no free lunch. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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Stephanie- There is a protocol for embedding cultured cell layers on our Center for Microscopy & Microanalysis web site. You should skip the step of washing with PBS; it is hard to make this reproducible. Just throw off the medium and pour on the fixative. You recover the cells in what is basically a cast of the cell culture. Ou should get a smooth, glass-like surface where the epoxy resin made contact with the cells. It is hard to cut out the little pieces, but MUCH easier than centrifuging cells down after every step. Antoher advantage is that you keep the relationship of the cells with one another and can see the cell layers (if any), intercellular junctions, etc.
The center web site should appear at the signature line.
==============================Original Headers============================== 4, 15 -- From heckman-at-bgnet.bgsu.edu Wed Mar 8 12:15:35 2006 4, 15 -- Received: from smtp01.bgsu.edu (smtp01.bgsu.edu [129.1.5.17]) 4, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28IFVZC025545 4, 15 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 12:15:33 -0600 4, 15 -- Received: from localhost (webmail.bgsu.edu [129.1.5.22]) 4, 15 -- by smtp01.bgsu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k28HLieE020494; 4, 15 -- Wed, 8 Mar 2006 12:21:44 -0500 (EST) 4, 15 -- MIME-Version: 1.0 4, 15 -- Date: Wed, 8 Mar 2006 12:21:44 -0500 4, 15 -- From: "heckman-at-bgnet.bgsu.edu" {heckman-at-bgnet.bgsu.edu} 4, 15 -- Message-Id: {1141838504-16113.023.77-smmsdV2.1.2-at-smtp.bgsu.edu} 4, 15 -- X-SMMS-Source: 72.240.90.60 4, 15 -- To: {microscopy-at-microscopy.com} 4, 15 -- Cc: {nizets2-at-yahoo.com} 4, 15 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM ==============================End of - Headers==============================
The higher the voltage, the lower the contrast. For ultra-thin sections of biological material a voltage of 60-80 keV is best.
Stephane
--- nomy_nay-at-hotmail.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question was submitted to Ask-A-Microscopist by } (nomy_nay-at-hotmail.com) } from } http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } on Tuesday, March 7, 2006 at 22:09:53 } Remember to consider the Grade/Age of the student } when considering the Question } --------------------------------------------------------------------------- } Please reply to both nomy_nay-at-hotmail.com as well } as to the Microscopy Listserver } --------------------------------------------------------------------------- } } Email: nomy_nay-at-hotmail.com } Name: Naomi Piyaratna } } Organization: Wollongong University, Australia } } Education: Undergraduate College } } Location: Wollongong, NSW, AUSTRALIA } } Title: Electron Miscroscopy. } } Question: In electron microscopy, the higher the } voltage the greater the penetrating abilityof the } electron beam, but the trade is a reduction of what? } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 8, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Mar } 8 08:40:26 2006 } 8, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 8, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k28EeP1i016821 } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 } Mar 2006 08:40:26 -0600 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } (Unverified) } 8, 12 -- Message-Id: } {p0611040bc0349d3016f2-at-[206.69.208.22]} } 8, 12 -- Date: Wed, 8 Mar 2006 08:40:25 -0600 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: nomy_nay-at-hotmail.com (by way of } Ask-A-Microscopist) } 8, 12 -- Subject: AskAMicroscopist: higher voltage } decreases what? } 8, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 7, 20 -- From nizets2-at-yahoo.com Wed Mar 8 11:30:43 2006 7, 20 -- Received: from web37404.mail.mud.yahoo.com (web37404.mail.mud.yahoo.com [209.191.87.57]) 7, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28HUYrV009464 7, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 11:30:35 -0600 7, 20 -- Received: (qmail 68044 invoked by uid 60001); 8 Mar 2006 17:02:30 -0000 7, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 20 -- s=s1024; d=yahoo.com; 7, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 20 -- b=PShnOhKIJY78JybHM2/w+l8sinFMsUxsKDTKohPBgoXeZU+rwzqvRU6vXUYf+jGrPARL/y3UDMKQ4T/ZNCoJLw6rIBR/vdumfn3A1wR30+ha5vOLnaL3CB/my9qlOjD8ZNTpXDoVusBtGh570vupwwCltrkfv8qtKgJfg22xQqo= ; 7, 20 -- Message-ID: {20060308170230.68042.qmail-at-web37404.mail.mud.yahoo.com} 7, 20 -- Received: from [80.122.101.102] by web37404.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 09:02:30 PST 7, 20 -- Date: Wed, 8 Mar 2006 09:02:30 -0800 (PST) 7, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: higher voltage decreases what? 7, 20 -- To: nomy_nay-at-hotmail.com 7, 20 -- Cc: microscopy-at-microscopy.com 7, 20 -- In-Reply-To: {200603081633.k28GXqOZ021992-at-ns.microscopy.com} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; charset=iso-8859-1 7, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Dear Naomi, I don't know how many others have replied to your inquiry. The larger beam-sample interaction volume that results from a higher beam voltage results in the signal coming from deeper in the sample, rather than just from the surface. This gives information from deeper in the sample, but sacrifices information from the very surface. If you need to see small features on the surface of your sample, a lower accelerating voltage is better. I hope this helps. Any basic SEM text should cover this point. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- X-from: {nomy_nay-at-hotmail.com} To: {mager-at-interchange.ubc.ca} Sent: Wednesday, March 08, 2006 7:57 AM
The ongoing discussion about contrast brings to mind another question. If one wants to add enough heavy metal to label a singular structure on a biological tissue thin section, how much metal is required to obtain a useful signal on a standard TEM? Would a STEM system allow one to "see" the structure with a lower amount of heavy metal label? Or does an energy filtered electron microscope (like the Zeiss 902) permit one to resolve smaller clusters?
I remember that some gold-linked antibody probes used fairly small gold clusters (11 atoms perhaps?) to improve penetration into the section, but that these ABs were only made visible after silver enhancement for routine TEM. When does a cluster of metal atoms become resolvable in a minimally stained thin section? -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
==============================Original Headers============================== 4, 21 -- From hall-at-aecom.yu.edu Wed Mar 8 14:06:39 2006 4, 21 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 4, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28K6c8f026310 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 14:06:38 -0600 4, 21 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 4, 21 -- by mailgw.aecom.yu.edu (8.12.11/8.12.11) with SMTP id k28K6WnP016413 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 15:06:38 -0500 4, 21 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 4, 21 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006030815063806598 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 08 Mar 2006 15:06:38 -0500 4, 21 -- Received: from [129.98.90.160] (worm.aecom.yu.edu [129.98.90.160]) 4, 21 -- by post.aecom.yu.edu (Postfix) with ESMTP id EB5852FD5 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 15:06:37 -0500 (EST) 4, 21 -- Mime-Version: 1.0 4, 21 -- X-Sender: hall-at-mailserver.aecom.yu.edu 4, 21 -- Message-Id: {a05100306c034e75199ac-at-[129.98.90.160]} 4, 21 -- Date: Wed, 8 Mar 2006 15:06:35 -0500 4, 21 -- To: microscopy-at-microscopy.com 4, 21 -- From: David Hall {hall-at-aecom.yu.edu} 4, 21 -- Subject: contrast and imaging small clusters of heavy metals 4, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
} Hi Sephanie Can you use glass coverslips? If so } I have a protocol I can send you where you can } embed the whole coverslip (cell side down) in a } chang embedding mold. You then remove the glass } using hydrofluoric acid, punch out small resin } circles of cells using a leather punch . Then } re-attach on to a blank resin stub and } section. That way you can get many blocks from } 1 coverslip. As mentioned earlier, avoid the PBS } step and scraping as both can destroy the morphology.
JoAnn Buchanan Stanford University School of Medicine
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
==============================Original Headers============================== 9, 20 -- From redhair-at-stanford.edu Wed Mar 8 14:42:18 2006 9, 20 -- Received: from smtp3.Stanford.EDU (smtp3.Stanford.EDU [171.67.16.138]) 9, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28KgHQt003672 9, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 14:42:17 -0600 9, 20 -- Received: from Bucky.stanford.edu (B135-WinXP.Stanford.EDU [171.65.21.62]) 9, 20 -- by smtp3.Stanford.EDU (8.12.11/8.12.11) with ESMTP id k28JaO6K016189; 9, 20 -- Wed, 8 Mar 2006 11:36:24 -0800 9, 20 -- Message-Id: {6.2.5.6.2.20060308112933.07445d60-at-stanford.edu} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 9, 20 -- Date: Wed, 08 Mar 2006 11:36:21 -0800 9, 20 -- To: nizets2-at-yahoo.com 9, 20 -- From: JoAnn Buchanan {redhair-at-stanford.edu} 9, 20 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 9, 20 -- Cc: "Microscopy Listserver" {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200603081644.k28Gip2n025969-at-ns.microscopy.com} 9, 20 -- References: {200603081644.k28Gip2n025969-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 9, 20 -- Content-Transfer-Encoding: 8bit 9, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k28KgHQt003672 ==============================End of - Headers==============================
Greater high voltage in a TEM is one of the few things in nature that does not have a lot of serious "Cons" that outweigh or balance the "Pros." Granted that increased radiation concerns and somewhat less contrast attend increasing the voltage, but on the plus side, the increased penetration, easier specimen preparation, improved resolution, plus others pros are BIG advantages.
Please forgive me if I point out that should you have a radiation sensitive specimen that you can always lower the voltage on a 300keV TEM for that specimen, but you can't raise the voltage on a 100keV machine to allow you to see through a thick specimen.
Ron Anderson
drteddunne-at-yahoo.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello Naomi, } } Tempting to say simply that the trade is a reduction } of contrast. That answer would apply if you were } asking about the trade of switching from say 40KV to } 100KV on the same microscope. } } You can compensate for this by using a smaller } objective aperture. } } If you are comparing a standard 40 to 100 KV } microscope with a high voltage mocroscope then the } answer is less straightforward and it is not } necessarily so that you have a serious contrast } reduction. Perhaps an electron microscope manufacturer } will see this question and give you more specific } answers. } } Best wishes, } } } Ted Dunn } The EMscope Company Ltd. } --- nomy_nay-at-hotmail.com wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The } } Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } This Question was submitted to Ask-A-Microscopist by } } (nomy_nay-at-hotmail.com) } } from } } } } } http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } } } on Tuesday, March 7, 2006 at 22:09:53 } } Remember to consider the Grade/Age of the student } } when considering the Question } } } } } --------------------------------------------------------------------------- } } } Please reply to both nomy_nay-at-hotmail.com as well } } as to the Microscopy Listserver } } } } } --------------------------------------------------------------------------- } } } Email: nomy_nay-at-hotmail.com } } Name: Naomi Piyaratna } } } } Organization: Wollongong University, Australia } } } } Education: Undergraduate College } } } } Location: Wollongong, NSW, AUSTRALIA } } } } Title: Electron Miscroscopy. } } } } Question: In electron microscopy, the higher the } } voltage the greater the penetrating abilityof the } } electron beam, but the trade is a reduction of what? } } } } } } } --------------------------------------------------------------------------- } } } } } ==============================Original } } Headers============================== } } 8, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Mar } } 8 08:40:26 2006 } } 8, 12 -- Received: from [206.69.208.22] } } (mac22.zaluzec.com [206.69.208.22]) } } 8, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } } ESMTP id k28EeP1i016821 } } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 } } Mar 2006 08:40:26 -0600 } } 8, 12 -- Mime-Version: 1.0 } } 8, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } } (Unverified) } } 8, 12 -- Message-Id: } } {p0611040bc0349d3016f2-at-[206.69.208.22]} } } 8, 12 -- Date: Wed, 8 Mar 2006 08:40:25 -0600 } } 8, 12 -- To: microscopy-at-microscopy.com } } 8, 12 -- From: nomy_nay-at-hotmail.com (by way of } } Ask-A-Microscopist) } } 8, 12 -- Subject: AskAMicroscopist: higher voltage } } decreases what? } } 8, 12 -- Content-Type: text/plain; } } charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? 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Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 10, 20 -- From drteddunne-at-yahoo.com Wed Mar 8 10:47:40 2006 } 10, 20 -- Received: from web33401.mail.mud.yahoo.com (web33401.mail.mud.yahoo.com [68.142.206.133]) } 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28Glche027028 } 10, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 10:47:39 -0600 } 10, 20 -- Received: (qmail 95631 invoked by uid 60001); 8 Mar 2006 16:47:38 -0000 } 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 10, 20 -- s=s1024; d=yahoo.com; } 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 10, 20 -- b=xpdlSSAAs0maYw04oUmPXHuqrXZmk07hPoZ8sv26sWh9auopxV0y/Sk8pC2kl0jDWpfW2TuOMk+g98SFzYOPojX2MrElhArfxv/3WaIY53wt88EQsbbUfuabYnRZpoWvdipk4Hgzqjz6poJVOSsr+FJE9qaEYUtJotg0YNB7sP8= ; } 10, 20 -- Message-ID: {20060308164738.95629.qmail-at-web33401.mail.mud.yahoo.com} } 10, 20 -- Received: from [202.47.247.156] by web33401.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 08:47:38 PST } 10, 20 -- Date: Wed, 8 Mar 2006 08:47:38 -0800 (PST) } 10, 20 -- From: ted dunn {drteddunne-at-yahoo.com} } 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: higher voltage decreases what? } 10, 20 -- To: nomy_nay-at-hotmail.com } 10, 20 -- Cc: microscopy-at-microscopy.com } 10, 20 -- In-Reply-To: {200603081542.k28FgiZ6003132-at-ns.microscopy.com} } 10, 20 -- MIME-Version: 1.0 } 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 } 10, 20 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 19 -- From randerson20-at-tampabay.rr.com Wed Mar 8 15:12:46 2006 5, 19 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01-smtplb.tampabay.rr.com [65.32.5.131]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28LCjXY012167 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 8 Mar 2006 15:12:46 -0600 5, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 19 -- by ms-smtp-01.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k28HBtjp013043 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 8 Mar 2006 12:11:57 -0500 (EST) 5, 19 -- Message-ID: {440F1059.5060502-at-tampabay.rr.com} 5, 19 -- Date: Wed, 08 Mar 2006 12:11:53 -0500 5, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: higher voltage decreases what? 5, 19 -- References: {200603081650.k28Gofmh028153-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200603081650.k28Gofmh028153-at-ns.microscopy.com} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
First, washing cells and centrifuging them before fixing will cause some changes in the morphology. Usually the changes are hidden after the cells have been embedded in epoxy resin because the resin is so good at holding the damaged cells together. As a general rule, for TEM examination it is best to handle the cells as little as possible, even after fixation. I think there is an old Nature paper by Pernilla et al that clearly demonstrates a loss of protein from unfixed cells during centrifugation.
After all, we don't need to wash cells if we don't have to. We are electron microscopists and can easily face the challenge of differentiating what is in the cells from what is outside. Why wash all the outside stuff away unless it is absolutely necessary?
We process cell monolayers in dishes similar to those you use and when we want to examine a pellet we fix first by adding double strength fixative to the cells as they are growing, and at the temperature they are growing at. After all, it is well known that changing the temperature of the cells can cause significant changes within the cells.
Once the fixative is added we wait 30 sec and then carefully scrape the cells from the substrate. Instead of regular cell scrapers we use small pieces of carefully trimmed teflon, or even shaped orange sticks that have been soaked in buffer before use.
Each dish is fixed and scraped individually and the cells immediately transferred to a tube for pelleting. We use an Eppendorf centrifuge that we bring up to top speed and then immediately let run down. It is important that each dish is treated individually and not batch processed as is routine in most labs. The only problem with a 6-well dish is that such individual attention is not easy when all the culture wells are on the same plate.
Once the fixed cells have been pelleted, we then leave the pellets to cool on ice. At some point we will add fresh, single strength fixative so that the pellet continues fixing. What is interesting is that if you pellet the cells while they are fixing and leave them as a pellet, a very strong clump of cells is formed. The cells do not fall apart so there is no need to continue centrifuging after that one pelleting step, and the pellet can be cut into smaller pieces for easier dehydration, infiltration and embedding.
Your dehydration and embedding protocol is not really important. Different dehydration agents produce different results, and the resin you use will also affect staining and sectioning qualities.
One of the most important parts of the process is the amount of time you grow your cells before fixing them. We always try to let the cells grow for at least 3 days before using them. I know that is not always possible when looking at transient transfections and other short-lived experiments, but letting the cells grow really does make a difference to the morphology. Again, I think there is lots of old (pre-pdf era) literature to dig into on the effect of growth on morphology (more proteins are made by the cells).
I think the processing of monolayers have been nicely covered already by other contributors, but I can add one extra point. If you want to remove glass or Thermaox slides from an epoxy resin block using liquid nitrogen, then the method works best if the resin has not been fully polymerized. Put the blocks in the oven in the morning and try to remove the blocks later in the day when the resin has hardened a little.
Best regards,
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
} From: {nizets2-at-yahoo.com} } Reply-To: {nizets2-at-yahoo.com} } Date: Wed, 8 Mar 2006 11:29:58 -0600 } To: {pwebster-at-hei.org} } Subject: [Microscopy] protocol to prepare cells in culture for TEM } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear colleagues and listers, } } I have some basic very technical questions. I want } to prepare MCF-7, } HepG2 and Caco-2 cells for TEM observation. } My protocol involves the following: } - grow cells on 6W multiwell plates up to } 70-80% confluency } - Wash cells 1x with cold PBS } - Add 1 ml cold PBS and detach cells with a } cell scraper } - Collect the cells in a 1.5 ml eppendorf } tube, rince the } well 1x with 500µl PBS and merge the volumes ----| } total 1.5 ml } - Centrifuge at 4°C for 5 min at 1500 RPM } - Pipet out carefully the supernatant and } carefully add cold } Karnovsky fixative } - â‘. } } When I follow this protocol with these cells, only the } HepG2 give a } nice pellet, the other give a too small pellet, } Please could you share with me your opinion about } this protocol? } - Should I forget 6W wells and grow cells } on normal petri } dishes? I would like to avoid that because I plan to } prepare 24 } conditions in parallel and working with 24 petri } dishes will be a pain. } - Is the centrifugation sufficient? Should } I increase the } centrifugation speed? } - Do you have a working protocol for the } preparation of } these cells for morphological observation? } Actually growing these cells in a way that they } arrive at the same } confluency at the same time is a real challenge since } their growth rate } are very different. This means that at least one cell } line won‚t be at } optimal confluency at the time of the experiment. I } know it‚s a question } of experience, but I don‚t want to want 1 month before } starting this } experiment :-) } } Another question: in parallel to this protocol, I am } trying to } develop a protocol for the embedding of cell } monolayers. The first attempt } was not too bad, the embedding basically worked, but } when I try to detach } the Epon resin from the bottom of the petri dish (I } cutted a square } with a saw), the surface of the resin is not flat. In } addition I don‚t } know where to cut my pyramid since I don‚t see where } the cells are on the } resin surface. Any clue? } } Thank you in advance, } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 4, 18 -- From nizets2-at-yahoo.com Wed Mar 8 09:39:20 2006 } 4, 18 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.87.61]) } 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k28FdJEq001920 } 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:39:20 -0600 } 4, 18 -- Received: (qmail 58964 invoked by uid 60001); 8 Mar 2006 15:39:19 } -0000 } 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 4, 18 -- s=s1024; d=yahoo.com; } 4, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-T } ransfer-Encoding; } 4, 18 -- } b=lXojiS18reQ7VfhwrGChK+qQJQofBqFpgp/cSONNcQy2bC84v0/AoWGPKo1xcDx/gMrYHPQWxBDJ } EWLm3VNKtKpDE+bdVglPv2ioU2n9JFt+kEOxwZOc8SZsqyApi8Kww4Xs9H10q48DO/xeyLlUc9Qk8H } MsJ3x6uD+DRefo2+o= ; } 4, 18 -- Message-ID: {20060308153919.58962.qmail-at-web37408.mail.mud.yahoo.com} } 4, 18 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via } HTTP; Wed, 08 Mar 2006 07:39:19 PST } 4, 18 -- Date: Wed, 8 Mar 2006 07:39:19 -0800 (PST) } 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 4, 18 -- Subject: protocol to prepare cells in culture for TEM } 4, 18 -- To: microscopy-at-microscopy.com } 4, 18 -- MIME-Version: 1.0 } 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 4, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers==============================
==============================Original Headers============================== 20, 20 -- From PWebster-at-hei.org Wed Mar 8 16:55:57 2006 20, 20 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 20, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28Mts57009838 20, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 16:55:55 -0600 20, 20 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 20, 20 -- Wed, 8 Mar 2006 22:52:26 +0000 20, 20 -- User-Agent: Microsoft-Entourage/11.2.1.051004 20, 20 -- Date: Wed, 08 Mar 2006 14:55:52 -0800 20, 20 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 20, 20 -- From: "Webster, Paul" {PWebster-at-hei.org} 20, 20 -- To: {nizets2-at-yahoo.com} , {microscopy-at-microscopy.com} 20, 20 -- Message-ID: {C034A0F8.8A9C%PWebster-at-hei.org} 20, 20 -- Thread-Topic: [Microscopy] protocol to prepare cells in culture for TEM 20, 20 -- Thread-Index: AcZDA3MAsXpDM672EdqlqAANk7Zh7g== 20, 20 -- In-Reply-To: {200603081729.k28HTvGp009203-at-ns.microscopy.com} 20, 20 -- Mime-version: 1.0 20, 20 -- Content-type: text/plain; 20, 20 -- charset="UTF-8" 20, 20 -- Content-Transfer-Encoding: 8bit 20, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k28Mts57009838 ==============================End of - Headers==============================
On Mar 8, 2006, at 7:17 AM, clei-at-uiuc.edu wrote:
} Higher voltage may } 1) Damage your samples easily. For Al foil, you can easily } see the beam damage at 300kv } 2) Cause multibeam effect when you use the diffraction } contrast techniques. Short wavelength means flat Ewards } sphere, or more beams are excited. } } OF course, cost and maintenance are also problems. } Dear Changhui & Naomi, All true, and one can take advantage of both. 1) Although the elastic and total cross sections decrease with increasing energy, the inelastic cross section rises with increasing energy, so by using an energy filter or collecting position-tagged spectra--a complete energy spectrum at each pixel obtained with the dose used for imaging--one can increase the contrast by using only the unscattered and elastically scattered electrons to produce the image, and one can collect (almost) all the electrons incident on the specimen and make use of their energy losses to identify the constituents; i.e., do element mapping. 2) Acquiring the higher-order diffraction information will allow one to get higher resolution, and diffraction contrast techniques are not the only case where this is true. I can't speak to the cost problem, but having maintained a 1.2 MeV HVEM for many years, I can say that, while more difficult than just buying a service contract, a dedicated staff can maintain good performance from such an instrument for decades. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Mar 8 17:00:43 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28N0fbw011238 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 17:00:42 -0600 4, 22 -- Received: from localhost (fire-dog [192.168.1.4]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 72F153456A 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 15:00:41 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id A465235D52 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Mar 2006 15:00:40 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603081517.k28FHOHf026899-at-ns.microscopy.com} 4, 22 -- References: {200603081517.k28FHOHf026899-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {ffad539d04d4f31bc0412cf77c665982-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: higher 4, 22 -- Date: Wed, 8 Mar 2006 15:09:15 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Hi Stephane I would not scrape cells until you have fixed them.
As to flat embedding cells; grow cells on Aclar film (EMS or Pella) fix and osmificate on the film process the aclar film just as you would any other prep embed and bake the plastic the film will still soft and will peal off, leaving a smooth plastic block with the cells in the plastic because the cells are just on the surface, and the block is smooth, and the cells are osmificated, they can easily be seen under a microscope I then take a fine saw and cut rectangular blocks out of the plastic and mount them in the microtome. This works well for X-sections of your cells. If you wish to section in the plane of the film, cut out small pieces and glue them (cell side up) to a blank
If you have any questions, contact me off-list. I probably have a protocol sitting around I could send you. David
On Mar 8, 2006, at 10:05 AM, nizets2-at-yahoo.com wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Dear colleagues and listers, } } I have some basic very technical questions. I want } to prepare MCF-7, } HepG2 and Caco-2 cells for TEM observation. } My protocol involves the following: } - grow cells on 6W multiwell plates up to } 70-80% confluency } - Wash cells 1x with cold PBS } - Add 1 ml cold PBS and detach cells with a } cell scraper } - Collect the cells in a 1.5 ml eppendorf } tube, rince the } well 1x with 500µl PBS and merge the volumes ----| } total 1.5 ml } - Centrifuge at 4°C for 5 min at 1500 RPM } - Pipet out carefully the supernatant and } carefully add cold } Karnovsky fixative } - } . } } When I follow this protocol with these cells, only the } HepG2 give a } nice pellet, the other give a too small pellet, } Please could you share with me your opinion about } this protocol? } - Should I forget 6W wells and grow cells } on normal petri } dishes? I would like to avoid that because I plan to } prepare 24 } conditions in parallel and working with 24 petri } dishes will be a pain. } - Is the centrifugation sufficient? Should } I increase the } centrifugation speed? } - Do you have a working protocol for the } preparation of } these cells for morphological observation? } Actually growing these cells in a way that they } arrive at the same } confluency at the same time is a real challenge since } their growth rate } are very different. This means that at least one cell } line won’t be at } optimal confluency at the time of the experiment. I } know it’s a question } of experience, but I don’t want to want 1 month before } starting this } experiment :-) } } Another question: in parallel to this protocol, I am } trying to } develop a protocol for the embedding of cell } monolayers. The first attempt } was not too bad, the embedding basically worked, but } when I try to detach } the Epon resin from the bottom of the petri dish (I } cutted a square } with a saw), the surface of the resin is not flat. In } addition I don’t } know where to cut my pyramid since I don’t see where } the cells are on the } resin surface. Any clue? } } Thank you in advance, } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 4, 18 -- From nizets2-at-yahoo.com Wed Mar 8 09:39:20 2006 } 4, 18 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.87.61]) } 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id } k28FdJEq001920 } 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 09:39:20 } -0600 } 4, 18 -- Received: (qmail 58964 invoked by uid 60001); 8 Mar 2006 } 15:39:19 -0000 } 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 4, 18 -- s=s1024; d=yahoo.com; } 4, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type: } Content-Transfer-Encoding; } 4, 18 -- } b=lXojiS18reQ7VfhwrGChK+qQJQofBqFpgp/cSONNcQy2bC84v0/AoWGPKo1xcDx/ } gMrYHPQWxBDJEWLm3VNKtKpDE+bdVglPv2ioU2n9JFt+kEOxwZOc8SZsqyApi8Kww4Xs9H1 } 0q48DO/xeyLlUc9Qk8HMsJ3x6uD+DRefo2+o= ; } 4, 18 -- Message-ID: } {20060308153919.58962.qmail-at-web37408.mail.mud.yahoo.com} } 4, 18 -- Received: from [80.122.101.102] by } web37408.mail.mud.yahoo.com via HTTP; Wed, 08 Mar 2006 07:39:19 PST } 4, 18 -- Date: Wed, 8 Mar 2006 07:39:19 -0800 (PST) } 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 4, 18 -- Subject: protocol to prepare cells in culture for TEM } 4, 18 -- To: microscopy-at-microscopy.com } 4, 18 -- MIME-Version: 1.0 } 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 4, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers==============================
==============================Original Headers============================== 8, 23 -- From Elliott-at-Arizona.edu Wed Mar 8 14:19:52 2006 8, 23 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 8, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28KJpwr030045 8, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2006 14:19:51 -0600 8, 23 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 8, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 8F665D3E079 8, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2006 13:19:50 -0700 (MST) 8, 23 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 8, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 7AC30D3B759 8, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2006 13:19:46 -0700 (MST) 8, 23 -- Mime-Version: 1.0 (Apple Message framework v623) 8, 23 -- In-Reply-To: {200603081705.k28H54vS000881-at-ns.microscopy.com} 8, 23 -- References: {200603081705.k28H54vS000881-at-ns.microscopy.com} 8, 23 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 8, 23 -- Message-Id: {c583c629128f46efca713255cec9301b-at-Arizona.edu} 8, 23 -- From: David Elliott {Elliott-at-Arizona.edu} 8, 23 -- Subject: Re: [Microscopy] protocol to prepare cells in culture for TEM 8, 23 -- Date: Wed, 8 Mar 2006 13:19:46 -0700 8, 23 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 8, 23 -- X-Mailer: Apple Mail (2.623) 8, 23 -- X-Virus-Scanned: amavisd-new at email.arizona.edu 8, 23 -- Content-Transfer-Encoding: 8bit 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k28KJpwr030045 ==============================End of - Headers==============================
There is another interesting approach, especially since you are working with c. Elegans. NT-MDT has just announced a new device which integrates an atomic force microscope with a Leica ultramicrotome. All of the early work has been done on either polymers or on c. elegans, especially by Dr. M. Mueller and Dr. N. Matsko at ETH in Zurich. The AFM uses local differences in elasticity and other physical properties to provide contrast, so in many cases, there is no need to stain with heavy metals. Also, because the AFM images from the block face, in sequential planes, it provides perfectly aligned sections for 3D reconstruction.
I understand that a demo unit will be available here in the States sometime around mid-year.
If you are interested in images and/or further information, please contact me off-line
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education www.MicroscopyEducation.com
313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018 ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). For details, call us at 972-943-8011 and ask for Ken Piel.
CAVEAT: MME is involved in the support of this techology and therefore has a financial interest.
At 03:14 PM 3/8/2006, hall-at-aecom.yu.edu wrote:
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==============================Original Headers============================== 14, 18 -- From bfoster-at-mme1.com Wed Mar 8 18:13:30 2006 14, 18 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 14, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k290DRk8029991 14, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 18:13:28 -0600 14, 18 -- Received: (qmail 27665 invoked from network); 8 Mar 2006 18:15:11 -0500 14, 18 -- Received: from host57-99.rancor.birch.net (HELO Barb-XP.mme1.com) (65.17.57.99) 14, 18 -- by enterprise.5starpro.com with SMTP; 8 Mar 2006 18:15:11 -0500 14, 18 -- Message-Id: {7.0.1.0.0.20060308170743.01d8d888-at-mme1.com} 14, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 18 -- Date: Wed, 08 Mar 2006 17:13:22 -0600 14, 18 -- To: hall-at-aecom.yu.edu, microscopy-at-microscopy.com 14, 18 -- From: Barbara Foster {bfoster-at-mme1.com} 14, 18 -- Subject: Re: [Microscopy] contrast and imaging small clusters of heavy 14, 18 -- metals 14, 18 -- In-Reply-To: {200603082114.k28LEj9f012730-at-ns.microscopy.com} 14, 18 -- References: {200603082114.k28LEj9f012730-at-ns.microscopy.com} 14, 18 -- Mime-Version: 1.0 14, 18 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
In the same vein as the complaints about having to go through SPAM filter certifications when sending listserver messages:::::::
Why is it that people do not "unsubscribe" from the listserver when they attend a meeting, go on vacation or do whatever it is they put in their auto-reply messages?
I get enough junk mail already so getting a barrage of these messages after I post on the listserver has one effect only - I don't post messages.
Now I wait for the auto-replies to arrive.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
==============================Original Headers============================== 9, 18 -- From PWebster-at-hei.org Wed Mar 8 18:57:36 2006 9, 18 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k290vY7T010073 9, 18 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 18:57:35 -0600 9, 18 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 9, 18 -- Thu, 9 Mar 2006 00:54:06 +0000 9, 18 -- User-Agent: Microsoft-Entourage/11.2.1.051004 9, 18 -- Date: Wed, 08 Mar 2006 16:57:33 -0800 9, 18 -- Subject: Out-of office replies 9, 18 -- From: "Webster, Paul" {PWebster-at-hei.org} 9, 18 -- To: {microscopy-at-microscopy.com} 9, 18 -- Message-ID: {C034BD7D.8AC0%PWebster-at-hei.org} 9, 18 -- Thread-Topic: Out-of office replies 9, 18 -- Thread-Index: AcZDFHK8sUqIzK8HEdqlqAANk7Zh7g== 9, 18 -- Mime-version: 1.0 9, 18 -- Content-type: text/plain; 9, 18 -- charset="US-ASCII" 9, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
This is most likely an Outlook issue. I use Eudora and simply set up an automatic kill filter for that subject (hence I changed this message's subject or I would not see my own posting--actually a good test). Your message's Subject does not exactly match Outlook's format so it did not get killed at my end.
Nestor has made numerous postings/Administrivia about this. I guess the problem still continues for those who don't have the ability to do filtering.
gary g.
At 05:42 PM 3/8/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Wed Mar 8 20:22:04 2006 9, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k292M3Y5001772 9, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 20:22:03 -0600 9, 20 -- Received: (qmail 11897 invoked from network); 8 Mar 2006 18:22:02 -0800 9, 20 -- Received: by simscan 1.1.0 ppid: 11893, pid: 11894, t: 0.1657s 9, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 20 -- by qsmtp1 with SMTP; 8 Mar 2006 18:22:02 -0800 9, 20 -- Message-Id: {6.2.3.4.2.20060308181546.02038dd0-at-mail.calweb.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 9, 20 -- Date: Wed, 08 Mar 2006 18:22:02 -0800 9, 20 -- To: PWebster-at-hei.org 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] Outofoffice replies 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200603090142.k291gfbj024539-at-ns.microscopy.com} 9, 20 -- References: {200603090142.k291gfbj024539-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I am been dismayed by the number of responses that answer the question as if it is for TEM.
Maybe I am missing something, but TEM is not mentioned in the question. Neither is the voltage range, such as 80KeV or 300KeV.
Hence, it is not obvious if the question relates to TEM, SEM, etc.....
My knee-jerk is response is: high penetration --- high transmission less reflection --- less surface sensitive
This would work equally well for SEM, STEM, TEM, AEM, etc......
JQuinn
PS: Neither is 'bio' vs 'materials'.
--------------------------------------------------------------------------- } } Email: nomy_nay-at-hotmail.com } Name: Naomi Piyaratna } } Organization: Wollongong University, Australia } } Education: Undergraduate College } } Location: Wollongong, NSW, AUSTRALIA } } Title: Electron Miscroscopy. } } Question: In electron microscopy, the higher the } voltage the greater the penetrating abilityof the } electron beam, but the trade is a reduction of what? } }
==============================Original Headers============================== 11, 12 -- From jquinn-at-www.matscieng.sunysb.edu Wed Mar 8 13:07:04 2006 11, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 11, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k28J6wCH009419 11, 12 -- for {microscopy-at-microscopy.com} ; Wed, 8 Mar 2006 13:07:01 -0600 11, 12 -- Received: (from jquinn-at-localhost) 11, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id k28J5Q113161 11, 12 -- for microscopy-at-microscopy.com; Wed, 8 Mar 2006 14:05:26 -0500 11, 12 -- Date: Wed, 8 Mar 2006 14:05:26 -0500 11, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 11, 12 -- Message-Id: {200603081905.k28J5Q113161-at-www.matscieng.sunysb.edu} 11, 12 -- To: microscopy-at-microscopy.com 11, 12 -- Subject: re: higher voltage ==============================End of - Headers==============================
Warm thanks to everyone for taking the time to instruct me about their protocol. I received a lot of answers, and lots of great ideas. Now I have to make a choice ;-)
Apparently general congruence can be observed for important steps: - Never wash with PBS, prefer direct fixation - Avoid scraping live cells. For this point one can ask why cell scrapers exist if they are so damaging to live cells. - There is still a possibility to use propyleneoxide in petri dishes, though it requires practice. I will keep this possibility as a "last resource" if nothing else works ;-) - Detaching the resin from the support after 12h (before complete curing) helps. - Growing cells in 6W format is definitely possible ;-) (which is a great new)
Some great ideas I will follow: - Using cut BEEM capsules during embedding of monolayers - Carbon-coating glass slides (I mean this one is really great isn't it?) - Fixing cells in the medium for a short time, collecting and centrifuging, then continuing fixation on the pellet (it helps a lot since I abandoned in situ fixation because I had a loose pellet and then too few cells per section) - When processing monolayers, minimize the time of contact with "extracting" substance (dehydration)
I don't always need to know the orientation of the cells, and so it was great to receive ideas for both pellet and monolayer embedding.
P.S1: I got only one clue how to localize the cells after monolayer embedding. Other help would still be welcome.
P.S2: Stephane is a french name, and it is different from Stephanie :D
Finally, I wish good luck to Pat in the land of the Sauerkraut and the biggest beer drinkers of the world.
Warm regards,
Stephane
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Sorry if I digress a bit, but I am new to the field of EDX. I thought that higher voltages gave a higher signal in EDX, and so a higher sensitivity. Is it not true?
Stéphane
http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi, } } In SEM, you will lose surface sensitivity. } } In TEM, scattering cross sections decrease which is } not good for EDX and EELS. } } Hongqi } } Dept. of Materials Science and Engineering } Pennsylvania State University } University Park, PA 16802 } email: hud105-at-psu.edu } } } ==============================Original
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I wish to thank the many members of this list who replied to my rather poorly posed question about EM costs. All of the replies were informative and helpful. They came in two flavors. Some wrote to say that I did not provide enough information to make even a rough guess, but they were kind enough to list the kinds of things I needed to specify or know in order to make cost estimates for these labs. Others described their labs and provided some cost estimates for their setups. I was also referred to some useful articles. I also had some replies from companies that sell new or used equipment, with information and prices. All told, I had about 2 dozen replies and followups in the two weeks following the initial request.
--aryeh -- Aryeh Weiss School of Engineering Bar Ilan University Ramat Gan 52900 Israel
Ph: 972-3-5317638 FAX: 972-3-5340697
==============================Original Headers============================== 4, 29 -- From aryeh-at-cc.huji.ac.il Thu Mar 9 04:09:16 2006 4, 29 -- Received: from tamar.os.biu.ac.il (tamar.os.biu.ac.il [132.70.60.24]) 4, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29A9FBQ015738 4, 29 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 04:09:15 -0600 4, 29 -- Received: from ismss-1.biu.ac.il (ismss.biu.ac.il [132.70.84.150]) 4, 29 -- by tamar.os.biu.ac.il (8.12.11/8.12.11/BIU) with ESMTP id k29A99KL696542 4, 29 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 12:09:13 +0200 4, 29 -- Received: from cc.huji.ac.il ([132.70.133.31]RDNS failed) by 4, 29 -- ismss-1.biu.ac.il with InterScan Message Security Suite; Thu, 09 Mar 2006 4, 29 -- 12:10:40 +0200 4, 29 -- Message-ID: {440FFEBF.40509-at-cc.huji.ac.il} 4, 29 -- Date: Thu, 09 Mar 2006 12:09:03 +0200 4, 29 -- From: Aryeh Weiss {aryeh-at-cc.huji.ac.il} 4, 29 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) 4, 29 -- Gecko/20030624 Netscape/7.1 (ax) 4, 29 -- X-Accept-Language: en-us, en, he 4, 29 -- MIME-Version: 1.0 4, 29 -- To: microscopy-at-microscopy.com 4, 29 -- Subject: cost of TEM, SEM and AFM 4, 29 -- Content-Type: text/plain; 4, 29 -- charset=us-ascii; 4, 29 -- format=flowed 4, 29 -- Content-Transfer-Encoding: 7bit 4, 29 -- X-imss-version: 2.038 4, 29 -- X-imss-result: Passed 4, 29 -- X-imss-scanInfo: M:P L:E SM:0 4, 29 -- X-imss-tmaseResult: TT:0 TS:0.0000 TC:00 TRN:0 TV:3.52.1006(14312.002) 4, 29 -- X-imss-scores: Clean:23.91834 C:2 M:3 S:5 R:5 4, 29 -- X-imss-settings: Baseline:2 C:1 M:1 S:1 R:1 (0.1500 0.1500) ==============================End of - Headers==============================
We are selling our practically unused cryo system for SEM, Oxford CT 1500B, bought in 1996, because of lack of space and suitable projects. The price is set to 5000 USD, which is far below the price of a new comparable system. The buyer will have to pay for the transportation. Please, ask for more information, if you are interested!
****************************** Kerstin Brismar B.Sc., Research engineer, Photographer Dept. of Crop Science SLU (Swedish University of Agricultural Sciences) P.O. Box 44 (Delivery: Växtskyddsvägen 1) SE-230 53 Alnarp, Sweden Phone: +46 40 41 55 05 Fax: +46 40 41 55 19 E-mail: Kerstin.Brismar-at-vv.slu.se ******************************
==============================Original Headers============================== 6, 17 -- From Kerstin.Brismar-at-vv.slu.se Thu Mar 9 04:33:27 2006 6, 17 -- Received: from alnus.slu.se (alnus.slu.se [194.47.49.5]) 6, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29AXQoK018422 6, 17 -- for {Microscopy-at-Microscopy.Com} ; Thu, 9 Mar 2006 04:33:26 -0600 6, 17 -- Received: from vv-238.vv.slu.se (vv-238.vv.slu.se [194.47.228.116]) 6, 17 -- by alnus.slu.se (8.12.11/8.12.11) with ESMTP id k29AXPYd021333 6, 17 -- for {Microscopy-at-Microscopy.Com} ; Thu, 9 Mar 2006 11:33:25 +0100 (CET) 6, 17 -- Message-Id: {6.2.5.6.0.20060309112709.01cbef08-at-vv.slu.se} 6, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 6, 17 -- Date: Thu, 09 Mar 2006 11:33:24 +0100 6, 17 -- To: Microscopy-at-Microscopy.Com 6, 17 -- From: Kerstin Brismar {Kerstin.Brismar-at-vv.slu.se} 6, 17 -- Subject: SEM - Cryo system for sale 6, 17 -- Mime-Version: 1.0 6, 17 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 6, 17 -- Content-Transfer-Encoding: 8bit 6, 17 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k29AXQoK018422 ==============================End of - Headers==============================
5 nm gold particles conjugated to antibodies are visible in "routine" TEM. Section staining doesn't necessarily have to be reduced to see the beads, although that can help. This isn't to say this is easy, but it can be done. A STEM or EELS would make the beads more identifiable, and zero-loss imaging in a TEM with EELs would mean very lightly stained, or unstained, OsO4 postfixed, sections could be examined. 3 nm might maybe just be doable, but I haven't tried. This is without any enhancement, Ag or otherwise. Highest spatial resolution is obtained if the gold particles are conjugated to primary antibodies.
Phil
} The ongoing discussion about contrast brings to mind another } question. If one wants to add enough heavy metal to label a singular } structure on a biological tissue thin section, how much metal is } required to obtain a useful signal on a standard TEM? Would a STEM } system allow one to "see" the structure with a lower amount of heavy } metal label? Or does an energy filtered electron microscope (like } the Zeiss 902) permit one to resolve smaller clusters? } } I remember that some gold-linked antibody probes used fairly small } gold clusters (11 atoms perhaps?) to improve penetration into the } section, but that these ABs were only made visible after silver } enhancement for routine TEM. When does a cluster of metal atoms } become resolvable in a minimally stained thin section? } -- } David H. Hall, Ph.D. } Center for C. elegans Anatomy } Department of Neuroscience } Albert Einstein College of Medicine } 1410 Pelham Parkway } Bronx, NY 10461 } } www.wormatlas.org } www.aecom.yu.edu/wormem } } phone 718 430-2195 } fax 718 430-2514 -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 24 -- From oshel1pe-at-cmich.edu Thu Mar 9 07:10:42 2006 4, 24 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29DAflY004701 4, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 07:10:41 -0600 4, 24 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 24 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k29Dn74l006366; 4, 24 -- Thu, 9 Mar 2006 08:49:07 -0500 4, 24 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 24 -- Thu, 9 Mar 2006 08:11:01 -0500 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {f06230900c035d8b5efa5-at-[141.209.160.132]} 4, 24 -- In-Reply-To: {200603082208.k28M82xx028864-at-ns.microscopy.com} 4, 24 -- References: {200603082208.k28M82xx028864-at-ns.microscopy.com} 4, 24 -- Date: Thu, 9 Mar 2006 08:10:37 -0500 4, 24 -- To: hall-at-aecom.yu.edu 4, 24 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 24 -- Subject: Re: [Microscopy] contrast and imaging small clusters of heavy 4, 24 -- metals 4, 24 -- Cc: Microscopy-at-microscopy.com 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-OriginalArrivalTime: 09 Mar 2006 13:11:01.0839 (UTC) FILETIME=[EA0C65F0:01C6437A] 4, 24 -- X-CanItPRO-Stream: default 4, 24 -- X-Spam-Score: -3.5 () L_EXCH_MF,PORN_RP_PENETRATIONS_ 4, 24 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Not strictly true... It depends on your examination goals. Here are two extreme, but not unusual, examples:
If I am looking for Pb somewhat deep below the surface (especially if the matrix contains S / Mo that can interfere with the low energy Pb lines), higher kV is indicated (20-30 kV). In this example I need the high kV to penetrate deeply and to excite the higher energy Pb lines. The higher energy Pb lines will better escape from the sample as well.
OTOH, if I am trying to identify micron size B4C crystals residing on a surface, low kV is indicated (2-5 kV). In this case, I desire low penetration to minimize excitation of the substrate and minimize dilution the response.
Regards, Woody White BWXT Services
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I too had noticed that many assumption this was a TEM question. Few addressed SEM although voltage is certainly an issue there.
I would also remind the other posters that this was submitted via Ask-a-Microscopist. There is an advisory on the initial post to remind us to copy the original inquirer on the discussion since they probably do not subscribe to the list. Some of you have been copying Naomi, but a lot haven't. She is missing out on your advice. It is easy to forget that as the discussion winds on.
Of course, we hope she will be happy enough with the responses that she becomes a regular list member.
Warren Straszheim Materials Analysis and Research Laboratory Iowa State University ------------------------------------------------ X-from: jquinn-at-www.matscieng.sunysb.edu [mailto:jquinn-at-www.matscieng.sunysb.edu] Sent: Wed 3/8/2006 1:35 PM To: wesaia-at-iastate.edu
Hi,
I understand. It sounds like against the common sense.
But the intensity of the EDX signal only depends on the element itself and the probability of scattering events. We use a factor " cross section" to quantified such probability. Look at its expression in any TEM book you will see the higher the voltage, the smaller the cross section.
Or I like to consider this question physically in the following way: Electrons can be consider as many single waves. The higher their voltage, the shorter their wavelength and the smaller the "size" of every of them. Apparently the small ball can travel longer in certain specimen. Just like a car is much easier to get blocked by the traffic than a motorcycle. Of course when there are no policemen. :-)
Hopefully this may help.
Hongqi
At 03:52 AM 3/9/2006, you wrote: } Sorry if I digress a bit, but I am new to the field of } EDX. I thought that higher voltages gave a higher } signal in EDX, and so a higher sensitivity. } Is it not true? } } Stéphane } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } Hi, } } } } In SEM, you will lose surface sensitivity. } } } } In TEM, scattering cross sections decrease which is } } not good for EDX and EELS. } } } } Hongqi } } } } Dept. of Materials Science and Engineering } } Pennsylvania State University } } University Park, PA 16802 } } email: hud105-at-psu.edu } } } } } } ==============================Original } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com
==============================Original Headers============================== 10, 23 -- From hud105-at-psu.edu Thu Mar 9 10:10:42 2006 10, 23 -- Received: from f04n07.cac.psu.edu (f04s07.cac.psu.edu [128.118.141.35]) 10, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29GAgWf003438 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 10:10:42 -0600 10, 23 -- Received: from Skyline.psu.edu ([128.118.37.73]) 10, 23 -- by f04n07.cac.psu.edu (8.13.2/8.13.2) with ESMTP id k29GAeTf028048; 10, 23 -- Thu, 9 Mar 2006 11:10:40 -0500 10, 23 -- Message-Id: {6.0.0.22.2.20060309102326.019b98f8-at-email.psu.edu} 10, 23 -- X-Sender: hud105-at-email.psu.edu (Unverified) 10, 23 -- X-Mailer: QUALCOMM Windows Eudora Version 6.0.0.22 10, 23 -- Date: Thu, 09 Mar 2006 11:09:15 -0500 10, 23 -- To: Stephane Nizet {nizets2-at-yahoo.com} 10, 23 -- From: Hongqi Deng {hud105-at-psu.edu} 10, 23 -- Subject: Re: higher voltages for EDX 10, 23 -- Cc: microscopy-at-microscopy.com 10, 23 -- In-Reply-To: {20060309085206.89288.qmail-at-web37405.mail.mud.yahoo.com} 10, 23 -- References: {200603082151.k28LpGbr023687-at-ns.microscopy.com} 10, 23 -- {20060309085206.89288.qmail-at-web37405.mail.mud.yahoo.com} 10, 23 -- Mime-Version: 1.0 10, 23 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 10, 23 -- X-Virus-Scanned: by amavisd-new 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k29GAgWf003438 ==============================End of - Headers==============================
Please don't make the mistake of simply correlating X-ray emission with a single parameter,like accelerating voltage, in either SEM or TEM applications.
Your measured x-ray intensity, as a function of accelerating voltage, is a product of a number of factors which include the ionization cross-section, electron beam current, electron energy loss, the scattering pathlength, and absorption path length.
As the accelerating voltage changes all of these parameters will vary and you need to include all of them in any assessment x-ray intensity for a given set of experimental conditions. The quantity that decreases with accelerating voltage is #Ionizations/nA/unitpathlength. Even though this quantity decreases with accelerating voltage for a constant probe size, it is likely that you will measure a higher x-ray signal as the accelerating voltage increases.
For example, below the critical excitation energy (Ec) for a given shell the x-ray emission for an element will be zero. It then increases rapidly to a broad maximum somewhere at ~ 2-4x Ec. Once you exceed ~ 4*Ec there is indeed a decrease in the cross-section. However this decrease is NOT linear, nor is it inversely proportional to accelerating voltage. Instead it is inversely proportional to the relativistically corrected energy of the electrons (1/2 mv^2), this means the decrease is not as great as you would expect. In addition, a number of electron sources actually yield higher beam currents at higher accelerating voltages, so even though the cross-section will be decreasing somewhat with accelerating voltage the net effect can be an increased x-ray signal, until such time as the depth of production is so great that the x-ray are absorbed within the sample before being detected.
If your bored and interested in seeing more detail on this for the TEM area, as well as some of the corresponding background and equations, go to the following URL
http://tpm.amc.anl.gov/Lectures/
then download the PDF file
XEDSAEMShortCourse.pdf
and look at pages 30-33 & 44-65.
Of course there are other deliterious effects of higher accelerating voltage, but that is a different discussion entirely.
Nestor Your Friendly Neighborhood SysOp
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
===========================================
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I understand your explanation, but the intensity of the signal (Y axis) in EDX does not depend on the nature of the material (this is the X axis), but on the number of times the same signal is read. This means that the intensity of the signal read by EDX depends on the number of electrons which hit a certain point on the sample, per unit of time. And this depends on the current. And least that's what I thought! :-D
Stéphane
--- Hongqi Deng {hud105-at-psu.edu} wrote:
} Hi, } } I understand. It sounds like against the common } sense. } } But the intensity of the EDX signal only depends on } the element itself and } the probability of scattering events. We use a } factor " cross section" to } quantified such probability. Look at its expression } in any TEM book you } will see the higher the voltage, the smaller the } cross section. } } Or I like to consider this question physically in } the following way: } Electrons can be consider as many single waves. The } higher their voltage, } the shorter their wavelength and the smaller the } "size" of every of them. } Apparently the small ball can travel longer in } certain specimen. Just like } a car is much easier to get blocked by the traffic } than a motorcycle. Of } course when there are no policemen. :-) } } Hopefully this may help. } } Hongqi } } } At 03:52 AM 3/9/2006, you wrote: } } Sorry if I digress a bit, but I am new to the field } of } } EDX. I thought that higher voltages gave a higher } } signal in EDX, and so a higher sensitivity. } } Is it not true? } } } } Stéphane } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } ---------------------------------------------------------------------------- } } } } } } Hi, } } } } } } In SEM, you will lose surface sensitivity. } } } } } } In TEM, scattering cross sections decrease which } is } } } not good for EDX and EELS. } } } } } } Hongqi } } } } } } Dept. of Materials Science and Engineering } } } Pennsylvania State University } } } University Park, PA 16802 } } } email: hud105-at-psu.edu } } } } } } } } } ==============================Original } } } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam } protection around } } http://mail.yahoo.com } }
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==============================Original Headers============================== 8, 20 -- From nizets2-at-yahoo.com Thu Mar 9 11:24:18 2006 8, 20 -- Received: from web37407.mail.mud.yahoo.com (web37407.mail.mud.yahoo.com [209.191.87.60]) 8, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k29HOHJk015444 8, 20 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 11:24:17 -0600 8, 20 -- Received: (qmail 54719 invoked by uid 60001); 9 Mar 2006 16:52:12 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=3kV/Nj+ncCCzrH6eYWFGstUn2UqHFAmOL9udYdlBDwVnGDQeQu0C+uCgHk+DzZS9fIs/tD4mn2DLSy/94rPK7qwUq4XX4ed7Wn9d3pwQj6OaHFg9fyGw53GF194WbW+FjG46qDCbVLZhe5sv4nedawZwRsQfrDNqWRNfn5EEAa0= ; 8, 20 -- Message-ID: {20060309165212.54717.qmail-at-web37407.mail.mud.yahoo.com} 8, 20 -- Received: from [80.122.101.102] by web37407.mail.mud.yahoo.com via HTTP; Thu, 09 Mar 2006 08:52:12 PST 8, 20 -- Date: Thu, 9 Mar 2006 08:52:12 -0800 (PST) 8, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 20 -- Subject: Re: higher voltages for EDX 8, 20 -- To: Hongqi Deng {hud105-at-psu.edu} 8, 20 -- Cc: microscopy-at-microscopy.com 8, 20 -- In-Reply-To: {6.0.0.22.2.20060309102326.019b98f8-at-email.psu.edu} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
You are partially correct. You are confusing total number of counts (which is what you are talking about) with the physics of x-ray generation, which is dependant upon both material and experimental conditions.
Counting longer only improves the statistics it will not increase the number of x-rays per electron per unit pathlength.
Nestor Your Friendly Neighborhood SysOp
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
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==============================Original Headers============================== 11, 16 -- From zaluzec-at-aaem.amc.anl.gov Thu Mar 9 11:33:20 2006 11, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 11, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29HXJC9018026 11, 16 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 11:33:19 -0600 11, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 11, 16 -- by aaem.amc.anl.gov (8.12.11/8.12.10) with ESMTP id k29HXJ61001218 11, 16 -- for {microscopy-at-microscopy.com} ; Thu, 9 Mar 2006 11:33:19 -0600 11, 16 -- Mime-Version: 1.0 11, 16 -- Message-Id: {p06110405c03616f433bc-at-[146.139.72.105]} 11, 16 -- Date: Thu, 9 Mar 2006 11:33:18 -0600 11, 16 -- To: microscopy-at-microscopy.com 11, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 11, 16 -- Subject: Re: higher voltages for EDX 11, 16 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 11, 16 -- Content-Transfer-Encoding: 8bit 11, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k29HXJC9018026 ==============================End of - Headers==============================
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Email: pedromfjcosta-at-gmail.com Name: Pedro Costa
Organization: University of Cambridge
Title-Subject: [Filtered] EDX
Question: I have been using the ES-Vision software, from EMISPEC, to do some quantitative analysis of STEM-EDX spectra taken with a 200 kV microscope. Besides allowing the input of experimental parameters (energy resolution, sample thickness / orientation, etc), this software is capable to do background subtraction, peak fitting, modelling of spectra and extract quantitative details for each element. As regards the quantification procedure, the final table of results shows several parameters such as weight percentage composition, atomic percentage composition and an uncertainty percentage. In case someone has used this software for the same purposes, would it be possible to clarify how does ES-Vision work out the uncertainties in the calculations?Also, what are the formulas used for the elemental percentages calculations (weight and atomic)?
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Email: mbisher-at-princeton.edu Name: Margaret Bisher
Organization: Princeton University
Title-Subject: [Filtered] Light Microscopy of Wood
Question: Hello Group,
I have a graduate student here who wants to image wood. It seems simple, but to her (and me, being a TEM person) it's not. She only wants to do light microscopy, so thick sections only. We can do paraffin or plastic embedding, or even cryo (we have a cryostat as well as a microtome).
My question to the group, is what do we do? We tried a standard dehydration with ethanol and zylene into paraffin, but the paraffin did not seem to penetrate completely into the wood and when cutting, the wood seems to crumble and just not be what we are hoping for.
Any suggestions would be really great. I've never done plant material, so this is really foreign to me.
The sticks are really tiny, most of them are going to be anywhere from 2 to 5 mm in diameter, not huge stalks..... she ultimately would like cross sections of these.
Margaret E. Bisher Electron Microscopy Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113B Princeton, NJ 08544-1014 Office: (609) 258-7026 Fax: (609) 258-8468 email: mbisher-at-molbio.princeton.edu
Get a book on botanical microtechnique before you do anything, it will save you a lot of wasted time. The 'woodies' I knew all used celloidion (parlodion) embedding.
Geoff
mbisher-at-princeton.edu wrote:
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What you do get at higher kV is a better peak-to-background ratio (at least in a TEM).
Characteristic X-rays are emitted isotropically. However, part of the background arises from bremstralung which is forward scattered (i.e. down the column) - the degree of forward scattering is dependent on the velocity of the electrons. Hence higher kVs result in the bremstralung forward scattering increasing. But, since the EDX background is not wholly dependent on bremstralung, the actual instrumental gain is not as much as you would expect from a simple physics argument.
In the case of SEM, you are probably best going to low kV, since this reduces the excitation volume, so improving the spatial resolution. However, this only really works with a FEG gun (to get enough probe current at low kV) and with WDX, since you have to work with L and M lines and need the resolution of WDX to separate the lines.
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
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Email: martimor-at-nmsu.edu Name: M. M.
Title-Subject: [Filtered] RE: propylene oxide
Question: I was wondering if anyone could answer a question that I have been wondering about? I was told by a technician by a certain company that acetone can act as a "scavenger" (okay?) when used as a transitional solvent for Araldite 502/Embed-812 medium and propylene oxide would always be better. Why would this be? I am seeking any advice on this matter please.
Does anyone in Australia have a video display unit suitable to fit a Hitachi H-600 TEM that they're willing to part with? Our second unit is having a near-death experience.
Thanks,
John Brealey Queen Elizabeth Hospital EM Unit IMVS - TQEH Pathology Adelaide, South Australia (08) 8222 6612
john.brealey-at-imvs.sa.gov.au
==============================Original Headers============================== 6, 35 -- From john.brealey-at-imvs.sa.gov.au Thu Mar 9 17:01:39 2006 6, 35 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 6, 35 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k29N1bnY032162 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 9 Mar 2006 17:01:38 -0600 6, 35 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 6, 35 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k29N1ZZr004923 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:35 +1030 (CST)' 6, 35 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 6, 35 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k29N1ZgP004906 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:35 +1030 (CST)' 6, 35 -- Received: from localhost (mesh.imvs.sa.gov.au [10.20.98.37]) 6, 35 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id EABF72EC03E 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:34 +1030 (CST) 6, 35 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 6, 35 -- by localhost (mesh.imvs.sa.gov.au [10.20.98.37]) (amavisd-new, port 10024) 6, 35 -- with LMTP id 31177-01-20 for {Microscopy-at-MSA.Microscopy.Com} ; 6, 35 -- Fri, 10 Mar 2006 09:31:34 +1030 (CST) 6, 35 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 6, 35 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id B81C92EC024 6, 35 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Mar 2006 09:31:34 +1030 (CST) 6, 35 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 6, 35 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 6, 35 -- Subject: TEM Hitachi H-600 6, 35 -- Date: Fri, 10 Mar 2006 09:31:34 +1030 6, 35 -- Message-ID: {000201c643cd$6979e270$c88a140a-at-iqe36042} 6, 35 -- MIME-Version: 1.0 6, 35 -- Content-Type: text/plain; 6, 35 -- charset="iso-8859-1" 6, 35 -- Content-Transfer-Encoding: 7bit 6, 35 -- X-Priority: 3 (Normal) 6, 35 -- X-MSMail-Priority: Normal 6, 35 -- X-Mailer: Microsoft Outlook 8.5, Build 4.71.2173.0 6, 35 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 6, 35 -- Importance: Normal 6, 35 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au ==============================End of - Headers==============================
I learned EM from John Luft around the time he pioneered use of Epon and propylene oxide. Four years later I diverted to acetone after being surprised and impressed by the results of ROBERTSON, J. D., BODENHEIMER, T. S., and STAGE, D. E., J. Cell Biol., 1963, 19, 159. Still later i tested 60 degree C heat-cure of 10 ml test-tubes filled with my Araldite 506 embedding mixture deliberately adulterated by inclusion of 15-20% solvent, trying propylene oxide, acetone, EtOH and MeOH. I found no reason then or since to give up acetone in favor of the others. Specifically, the propylene oxide mix cured to give a softer and somewhat cheesy polymer; the acetone mix reduced somewhat in volumed during cure and the cured product was similar to unadulterated resin. I can't recall the alcohols results; I think they were similar to acetone? . I was surprised that the propylene oxide result was inferior to the acetone result; I had expected both to evaporate during cure to leave a final polymer unaltered y the solvent inclusion.
I never heard the term "scavenger" applied. Luft called propylene oxide a "reactive diluent", and suggested that any small amount that failed to evaporate during heat-cure would incorporate harmlessly in the polymer. He was probably correct. My point is that large amounts of propylene oxide are not so "harmless", while similarly large amounts of included acetone are surprisingly innocuous. -mike reedy-
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-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
Hello, I was wondering if anyone had used the Navitar Video Fluorescence Scope: http://www.navitar.com/zoom/zfl_gen.htm
I was considering using it with some bacteria fluorescence probes and was wondering if anyone had tried it out and what they thought of it. Any advice appreciated.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
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I notice that Epson has just started shipping the V700 [and V750 Pro] large format flatbed scanner that costs around Ł400 to Ł550 [for the Pro version that has enhanced optics and Silverfast Ai] in the UK. The V700 is a top end consumer model and the V750 is a 'professional' model (and so offers more).
Check it out at: http://www.photo-i.co.uk/News/Feb06/Epson_V700_scanner.htm and click the 'interactive review'.
This scanner is clearly a real advance on the slightly cheaper Epson 4990F and Canon 9950F scanners that have 4,800 dpi. The V700 [and V750] is rated at 6,400 dpi. The new Epson V750 Pro produces sharper focus in its scans than these previous models that gave quite 'soft' images - this is no doubt due to better optics.
X-from the review link above it is clear that the V700 series scanner is a significant improvement on the last generation of prosumer flat bed scanners and should be a serious contender for any shortlist on those wishing to scan large format negatives/positives up to A4 in size (i.e. TEM negatives).
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
Is this site truely independent or is it the voice of the manufacturer or industry group? Has anyone taken a TEM negative and scanned it on the new v. the old scanners? And does anyone need a TEM negative scanned at 6400 dpi? I would rather see an real increase in the dynamic range rather than a dpi "race".
Geoff
keith.morris-at-ucl.ac.uk wrote:
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==============================Original Headers============================== 6, 32 -- From mcauliff-at-umdnj.edu Mon Mar 13 09:08:18 2006 6, 32 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 6, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2DF8IMg010097 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Mar 2006 09:08:18 -0600 6, 32 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 6, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id B4ECD4BE3D 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Mar 2006 09:08:17 -0600 (CST) 6, 32 -- Received: from polaris.umdnj.edu (polarisb.UMDNJ.EDU [130.219.34.133]) 6, 32 -- by zix02.umdnj.edu (Proprietary) with ESMTP id 869DD4BE31 6, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Mar 2006 09:08:16 -0600 (CST) 6, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 32 -- id {0IW200I01N6CAI-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 32 -- for microscopy-at-msa.microscopy.com; Mon, 13 Mar 2006 10:08:15 -0500 (EST) 6, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 6, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 32 -- 2004)) with ESMTP id {0IW200GSINCH7J-at-Polaris.umdnj.edu} ; Mon, 6, 32 -- 13 Mar 2006 10:07:31 -0500 (EST) 6, 32 -- Date: Mon, 13 Mar 2006 10:06:37 -0500 6, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 32 -- Subject: Re: [Microscopy] Scanner for TEM Micrographs 6, 32 -- In-reply-to: {200603131321.k2DDLqdn024056-at-ns.microscopy.com} 6, 32 -- To: keith.morris-at-ucl.ac.uk, 6, 32 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 32 -- Message-id: {44158A7D.3000902-at-umdnj.edu} 6, 32 -- MIME-version: 1.0 6, 32 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 32 -- Content-transfer-encoding: 8BIT 6, 32 -- X-Accept-Language: en-us, en 6, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 32 -- Gecko/20040804 Netscape/7.2 (ax) 6, 32 -- References: {200603131321.k2DDLqdn024056-at-ns.microscopy.com} ==============================End of - Headers==============================
Dear All John Benjamin Dancer produced photomicrographs of fleas in the mid-19th century. Was he the very first person to record a microscope image using photography?
Who was the first person to make a photomicrograph of a protein crystal?
Best wishes Chris
Dr Christopher E. Jeffree University of Edinburgh Institute of Molecular Plant Sciences King's Buildings, Mayfield Road Edinburgh, EH9 3JH Scotland, UK Tel: +44 131 650 5554 FAX: +44 131 650 5392 email c.jeffree-at-ed.ac.uk
==============================Original Headers============================== 5, 26 -- From c.jeffree-at-ed.ac.uk Mon Mar 13 10:04:44 2006 5, 26 -- Received: from lawnmarket.ucs.ed.ac.uk (lawnmarket.ucs.ed.ac.uk [129.215.166.63]) 5, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2DG4h0w024583 5, 26 -- for {microscopy-at-microscopy.com} ; Mon, 13 Mar 2006 10:04:43 -0600 5, 26 -- Received: from EMFBIO (emf.icmb.ed.ac.uk [129.215.156.56]) 5, 26 -- by lawnmarket.ucs.ed.ac.uk (8.12.10/8.12.10) with SMTP id k2DG4gNH022706 5, 26 -- for {microscopy-at-microscopy.com} ; Mon, 13 Mar 2006 16:04:42 GMT 5, 26 -- Message-ID: {000e01c646b7$51009530$389cd781-at-EMFBIO} 5, 26 -- Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} 5, 26 -- From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} 5, 26 -- To: {microscopy-at-microscopy.com} 5, 26 -- Subject: First photomicrograph 5, 26 -- Date: Mon, 13 Mar 2006 16:00:57 -0000 5, 26 -- MIME-Version: 1.0 5, 26 -- Content-Type: text/plain; 5, 26 -- format=flowed; 5, 26 -- charset="iso-8859-1"; 5, 26 -- reply-type=original 5, 26 -- Content-Transfer-Encoding: 7bit 5, 26 -- X-Priority: 3 5, 26 -- X-MSMail-Priority: Normal 5, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 5, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 5, 26 -- X-Edinburgh-Scanned: at lawnmarket.ucs.ed.ac.uk 5, 26 -- with MIMEDefang 2.33, Sophie, Sophos Anti-Virus 5, 26 -- X-Scanned-By: MIMEDefang 2.33 (www . roaringpenguin . com / mimedefang) ==============================End of - Headers==============================
To be honest most of us here use only 800 to 1,200 dpi for TEM negative scanning anyway, whether for publication or image analysis (at this dpi the Epson 9950F will no doubt produce similar scans to the new Epson V750). We may zoom in on areas for scanning (enlargements), but never 'archive' the whole TEM negative as digitised images - we just keep the negatives. Our new Epson 9950F flatbed scanner we use (cost Ł280 in 2006) produces digitised images scanned at maximum resolution that are distinguishable to those from our Agfa DuoScan flatbed scanner (that cost Ł5,000 in 2001). Both scanners were at their maximum usable resolution of 2,400 dpi and 2,500 dpi respectively. I have noticed that at 4800 dpi the Epson 4990 Photo has juddered once while scanning - this might be due to inferior mechanics or poor vibration protection, the heavy DuoScan has a 'squash ball' type isolation feature. The DuoScan is slower and noisier though, and can only scan one negative at a time in it's 'sweet spot', compare to three to six negatives with the Epson 4990 Photo (it's also SCSI rather than universal USB2). Image morphometry distortion is also very low across the platter with the Epson, producing mean errors in length and area measurements of around 0.3% at 800 dpi and 0.15% at 2,400 dpi (using graph paper as a target). This is very close to the inherent errors from using the mouse in MetaMorph.
Images from both scanners need a quick bit of Photoshop work to get them looking their best. We used to spend hours doing that when printing EM photographs in the dark room - now with these cheap multi-purpose flatbed scanners you should never have to do that again.TEM negatives go up to a maximum of around six times enlargement, although at high TEM magnifications the resin grain may be more of a problem than the film grain. I'm also scanning a few B&W TEM negatives and colour 35mm slides on UCL's 8000 dpi Imacon professional scanner at UCL's Reprographics for comparison - that's Ł5 to Ł10 (} 50Mb) per scan though depending on image file size.
DMax in these modern cheap flatbed scanners is reported to be around 3.8 to 4.0 (its difficult to compare specs though as, like with dpi, manufacturers lie about the true value differently). Correctly exposed B&W silver halide negatives are reported to have a DMax of nearer 1.5 compared to a colour dye slides 3.5D - so a good scanners DMax is less of a problem with B&W negatives and even a dog of a modern scanner should cope with most TEM negatives in terms of dynamic range. Plus we can only distinguish 191 grey levels, so 8-bit (256 greys) rather than 14-bit (16,385 greys) is mostly fine for B&W photographs - we do a bit better with colour and so need things like CMYK printing and ICC profiling for this. As with microscopes, fantastic resolution isn't much use if there's no contrast, but again as with microscopes, increased contrast often has the cost of reducing resolution. B&W TEM negatives that initially look good with very high contrast (DMax nearer 2.4) are generally inferior in detail to negatives that have a far more neutral tonal balance. One problem with TEM negatives is that we can't immediately tell if the picture is poor after zooming in, particularly if a cheap scanner secretly applies USM, whereas if its a colour scan of our kids faces, or writing on the side of a ship, it's immediately obvious. For TEM negatives it's easier just to compare the results from different scanners and with the manual view looking at the negative with a light box and a magnifier. Although we don't need a [probably optimistic] '6,000 dpi' for large format negatives unless we really want to look at the film grain, it does suggest that the scanner has very good optics for great lower resolution scans. The use of Digital ICE [FARE] dust removal is pretty irrelevant for B&W negatives as the process is optimised for colour film only.
It naturally tends to be photographers who want the higher resolutions of 4,000 dpi and above, plus sharp focussing, largely for the archiving smaller 35mm colour slide or negatives. Here resolving detail in shadow with low noise [i.e. high DMax] is very important - further helped by Photoshop CS's great 'shadow/highlight' feature. My colour slides of the family from the 1950's to 1980's are all showing signs of aging, in particular the 50's slides that have now gone very dark brown (although a few Fuji slides from the 70's have also really aged badly - fortunately I mainly used Afga/Perutz back then). I now wish I hadn't got myself the Canon 9950F (Ł260) for home use at Christmas - the new Epson V750 would have made a better job of digitising my family colour slides and photos (although I probably wouldn't notice the difference much at A4 after USM, and the V750 is Ł200 more). At work we are happy to continue with the Epson 4990 Photo, although I obviously would have bought the Epson V750 with its sharper optics if I was buying now - the Epson 4990 Photo and Canon 9950F do definitely produce 'softer' slightly out of focus scans (and their output quality is identical). Also don't buy the Canon 9950F to scan TEM negatives, it can't do 'A4' negative scans and is restricted to the sizes of the film holders (that are 4 x 5" and 120 and not standard EM negative sizes). The Epson 4990 (and the near identical V750) can scan negatives to nearly the full platter size - that means three or six TEM negatives in one go (depending on their size).The V750 can also scan more 35mm slides and negatives in one go than the 4990. We have plenty of 35mm film to scan here from old optical photo-microscopes, 'talks' and lab cameras. My Canon 9950F flatbed scanner at home produces better colour scan images, particularly from colour negatives, than my comparably priced dedicated Acer [Benq] ScanWit 2740s 2,700 dpi 35mm slide scanner - no doubt the Epson V750 flatbed would do even better.
I suppose we should use 'acid free' bag storage to protect our colour negatives, photographs and slides from atmospheric pollution and decay - just the same as the valuable linen, comic and book crowd do - but I've never got around to it. Fortunately time has demonstrated that the silver halide process produces a far more stable image, albeit black and white, compared to those produced with colour dyes. The support material though, e.g particularly old cellulose nitrate stock, may degrade badly with time. Early photographers fortunately used glass plate as the support medium that was very durable [until you drop them]. I have a few 1920's large format B&W film negatives of the family and 1930's16mm B&W movie film (Pathe News) that still look good though.
Keith
PS. The http://www.photo-i.co.uk site is an independent one run in the UK. The site is quite similar to the excellent http://www.dpreview.com for digital cameras. It's quite clear from their reviews that they are independent - although they are naturally keen photographers rather than electron microscope users so their priorities may differ. If they say some aspect of the product will really pig you off - it invariably does. I expect they do consultancy reviews for magazines and that manufacturers are keen to get them to review their product if they think it's good. It's also rather obvious that in short review articles in the like those of PCPro magazine [in the UK] the reviewers have often spent hardly any time trying to get the best out of the scanner.
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
My last posting missed out a rather key 'in' from 'in'distguishable (I should have stuck with the less technically correct term 'identical', as in the my first draft). The sentence in paragraph one, line three, thus should have read:
"Our new Epson 9950F flatbed scanner we use (cost Ł280 in 2006) produces digitised images scanned at maximum resolution that are indistinguishable to those from our Agfa DuoScan flatbed scanner (that cost Ł5,000 in 2001)."
i.e. both scanned images look pretty much the same and you can't tell them apart once you have put them through Photoshop's autocontrast adjustment. Prior to Photoshop editing, the 'auto' setting on the older Agfa DuoScan twain interface produced quite light scans that actually looked worse than the cheap Epson 9950F's more contrasty ones.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
I have always made my Epon with...well Epon. Now I come in a lab (i am the only EM here) where we have stocks of Glycid ether. Are they similar products? Should I use it the same way as Epon and at the same proportions?
Stephane (without i ;-))
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Tue Mar 14 10:42:38 2006 7, 18 -- Received: from web37408.mail.mud.yahoo.com (web37408.mail.mud.yahoo.com [209.191.87.61]) 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2EGgc7H026412 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 14 Mar 2006 10:42:38 -0600 7, 18 -- Received: (qmail 36782 invoked by uid 60001); 14 Mar 2006 16:42:38 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=SGVnigD362HyNBBpWqulINdx4DZuhpYEH/KXo23eBg0s9tANhKguMtkdM5HYPG2SwQBMv0F2knf6QxcjiN3EvhoqAmnK0AnEpPFdsd97p+/a88DqJ0yy4Zd5qFq+V89n7jiQo3B9Mq/vZjLSa5mK51I+Hv06akJlSUS8nczo9Mw= ; 7, 18 -- Message-ID: {20060314164238.36780.qmail-at-web37408.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via HTTP; Tue, 14 Mar 2006 08:42:38 PST 7, 18 -- Date: Tue, 14 Mar 2006 08:42:38 -0800 (PST) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: glycid Ether 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; charset=iso-8859-1 7, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
} -----Original Message----- } From: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Tuesday, March 14, 2006 10:50 AM } To: Dusevich, Vladimir } Subject: [Microscopy] glycid Ether } } } } } -------------------------------------------------------------- } -------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } -------------- } } Dear all, } } I have always made my Epon with...well Epon. Now I come in a } lab (i am the only EM here) where we have stocks of Glycid ether. } Are they similar products? Should I use it the same way as } Epon and at the same proportions? } } Stephane (without i ;-)) } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection } around http://mail.yahoo.com } } ==============================Original } Headers============================== } 7, 18 -- From nizets2-at-yahoo.com Tue Mar 14 10:42:38 2006 7, } 18 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.87.61]) } 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP } id k2EGgc7H026412 } 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 14 Mar } 2006 10:42:38 -0600 } 7, 18 -- Received: (qmail 36782 invoked by uid 60001); 14 Mar } 2006 16:42:38 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; } q=dns; c=nofws; } 7, 18 -- s=s1024; d=yahoo.com; } 7, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Conten t-Type:Content-Transfer-Encoding; } 7, 18 -- } b=SGVnigD362HyNBBpWqulINdx4DZuhpYEH/KXo23eBg0s9tANhKguMtkdM5HY } PG2SwQBMv0F2knf6QxcjiN3EvhoqAmnK0AnEpPFdsd97p+/a88DqJ0yy4Zd5qF } q+V89n7jiQo3B9Mq/vZjLSa5mK51I+Hv06akJlSUS8nczo9Mw= ; } 7, 18 -- Message-ID: } {20060314164238.36780.qmail-at-web37408.mail.mud.yahoo.com} } 7, 18 -- Received: from [80.122.101.102] by } web37408.mail.mud.yahoo.com via HTTP; Tue, 14 Mar 2006 } 08:42:38 PST 7, 18 -- Date: Tue, 14 Mar 2006 08:42:38 -0800 } (PST) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 } -- Subject: glycid Ether 7, 18 -- To: } microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- } Content-Type: text/plain; charset=iso-8859-1 7, 18 -- } Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== }
==============================Original Headers============================== 6, 23 -- From DusevichV-at-umkc.edu Tue Mar 14 11:29:02 2006 6, 23 -- Received: from KC-MSXPROTO2.kc.umkc.edu (pop3.exchange.umkc.edu [134.193.143.155] (may be forged)) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2EHT0qX002997 6, 23 -- for {microscopy-at-msa.microscopy.com} ; Tue, 14 Mar 2006 11:29:01 -0600 6, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by KC-MSXPROTO2.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Tue, 14 Mar 2006 11:28:59 -0600 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: RE: [Microscopy] glycid Ether 6, 23 -- Date: Tue, 14 Mar 2006 11:28:58 -0600 6, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADBF9-at-KC-MSX1.kc.umkc.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [Microscopy] glycid Ether 6, 23 -- Thread-Index: AcZHh08url+foB08SoSaavJe2GWPwQABWlcA 6, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 6, 23 -- To: {microscopy-at-msa.microscopy.com} 6, 23 -- X-OriginalArrivalTime: 14 Mar 2006 17:28:59.0657 (UTC) FILETIME=[C79CBF90:01C6478C] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2EHT0qX002997 ==============================End of - Headers==============================
The University of California at Santa Barbara (Molecular, Cellular, and Developmental Biology Department and the Neuroscience Research Institute) is offering a workshop on Advanced Microscopy Digital Imaging. This course is co-sponsored with Purdue University and will be held in Santa Barbara from April 24-28th. In addition to our academic sponsors, the workshop has the generous support of Media Cybernetics who is providing their software and support for a teaching assistant. Also, Olympus of America is providing four inverted microscope stands, a DSU (Disk Scanning Unit) and Fluoview scanning confocal microscope. There will be digital cameras from Q-Imaging, fluorescent filters from Omega Optical, cell injectors from Eppendorf, and stage heaters and live cell chambers from Bioptechs. For more information about the course, please check the following web site:
I have an Olympus BH-2 Microscope that requires a Polariser/Analyser set and filters for both light paths. I have been advised by the local Olympus reps that the microscope has been superseded many years ago and the parts are hard to come by.
Does anyone have any stashed in a draw that they would be willing to part with or know of any third party suppliers.
Any help would be greatly appreciated.
Regards George
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==============================Original Headers============================== 10, 27 -- From George.Theodossiou-at-amcor.com.au Tue Mar 14 18:02:05 2006 10, 27 -- Received: from aiti251.amcor.com.au (smtp.amcor.com.au [202.14.180.248]) 10, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2F022RK026584 10, 27 -- for {microscopy-at-msa.microscopy.com} ; Tue, 14 Mar 2006 18:02:03 -0600 10, 27 -- Received: from aadcex0001.amcor.net (unverified) by aiti251.amcor.com.au 10, 27 -- (Content Technologies SMTPRS 4.3.19) with ESMTP id 10, 27 -- {T7709ff7fcaa0de98c8bd0-at-aiti251.amcor.com.au} for 10, 27 -- {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 11:07:27 +1100 10, 27 -- Received: from aadcex0004.amcor.net ([160.222.80.235]) by 10, 27 -- aadcex0001.amcor.net with Microsoft SMTPSVC (6.0.3790.211); Wed, 15 Mar 10, 27 -- 2006 11:02:01 +1100 10, 27 -- Received: from 160.222.214.38 ([160.222.214.38]) by aadcex0004.amcor.net 10, 27 -- ([160.222.80.235]) with Microsoft Exchange Server HTTP-DAV; Wed, 15 Mar 10, 27 -- 2006 00:02:00 +0000 10, 27 -- User-Agent: Microsoft-Entourage/11.2.1.051004 10, 27 -- Date: Wed, 15 Mar 2006 11:01:25 +1100 10, 27 -- Subject: Filters For Olympus BH-2 10, 27 -- From: "George.Theodossiou" {George.Theodossiou-at-amcor.com.au} 10, 27 -- To: {microscopy-at-msa.microscopy.com} 10, 27 -- Message-ID: {C03DA485.101C%George.Theodossiou-at-Amcor.com.au} 10, 27 -- Thread-Topic: Filters For Olympus BH-2 10, 27 -- Thread-Index: AcZHw5m62ByWbLO2EdqaGwANkzYUMg== 10, 27 -- Mime-version: 1.0 10, 27 -- Content-type: text/plain; charset="US-ASCII" 10, 27 -- Content-transfer-encoding: 7bit 10, 27 -- X-OriginalArrivalTime: 15 Mar 2006 00:02:01.0242 (UTC) 10, 27 -- FILETIME=[AF550FA0:01C647C3] ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both maloneyb-at-fiu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: maloneyb-at-fiu.edu Name: Barbara
Organization: FIU
Title-Subject: [Filtered] How to change filament in Phillips 300 TEM
Question: I have changed filaments in other instruments, but never in this older model. The manual doesn't seem to cover this - does anyone have the written procedure. Really would appreciate it greatly. Thanks Barbara
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Email: pekysar-at-ucdavis.edu Name: Pat Kysar
Organization: University of California, Davis
Title-Subject: [Filtered] Low Dose TEM
Question: Hello all, We have a client who is trying to use the low dose function of our FEI CM120. He is looking at magnetic nano particles on a substrate. He was able to make it work but is still getting damage to his sample. Unfortuately, neither of the techs here have used the low dose so we are quite unfamiliar with it and aren't much help. He would like to know what radius he should be using and we would like to hear from anyone who has experience with the low dose on this tool (or any tips, for that matter) which might help him achieve success. We would all appreciate any help! Cheers, Pat Kysar University of California, Davis Medical School, Pathology EM Lab
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Email: biology-at-ucla.edu Name: Eric
Organization: UCLA Medical Center
Title-Subject: [Filtered] Cleaning Grids
Question: This was never a problem till about a month ago..
Since about a month we have been having a problem keeping the sections stuck to the grids. The grids are cleaned in 100% Acetone and dried. Sections are picked up either from above or below the water.
When the grids are stained using the microwave staining method we have been using for several years the sections come off the grids... Every now and then the sections will stick to the grids and everyhting is fine.
Any suggestions about consistency? I have tried staining less time in the microwave, but this does not make a difference...
Any different ways to clean the grids so the sections will stick better?
Dear Stephane, ... the problem with HV and 'sensitivity' of EDX measurements is very complex, as Nestor already stated. First, the overvoltage U = Eo/Ec (primary electron energy / critical shell ionisation energy) should be at least more than 2 for all elements of interest. If not, every gain in HV is leading to extreme excitation enhancements of the element (and X-ray production), like Nestor already stated. If U } 3, the X-ray excitation curve decreases quite flat.
If the basic excitation of characteristic X-rays is sufficient:
#1 } From the point of view of detection limits you have to consider the peak to background ratio. The background in EPMA is bremsstrahlung. The ratio of characteristic line excitation to bremsstrahlung excitation is always gaining with primary electron energy (HV). Therefore you achieve always better detection limits with higher HV (but see #2, the count rate limitation can work against!)
#2 You must take into account: Higher electron energy do always excite much more higher energy X-rays (both characteristic X-rays and bremsstrahlung). The count rate possibilities of an EDX are always limited. So you have to consider, that you possibly will get less count rates for the elements of interest with higher energy excitation... because your pulse processor has to process more high energy X-ray signals (less counts in a given time for the low energy X-rays you have in focus). E.g. The peaks are lower in a Au/Ag alloy between 2..4 keV (Ag-L/Au-M) with 40 kV excitation, than they would be with 15 kV. But the Au-L lines (near 10 keV) are 4 times higher (because 15 keV is very low for Au-L). Therefore, an increase of HV is bad for Ag-L/Au-M measurement, if a limited EDX count rate is given.
#3 Also the X-ray absorption in specimen is increasing with higher HV. Particularly weaker element counts will be very much reduced behind absorption jump of a main element. But to drive against absorption: Tilt the specimen towards the EDX detector. The absorption influence is not so important with TEM X-ray measurements (thin specimen).
Finally: The better detection limits ('sensitivity') goes really with higher HV. But one has to take into account limitations in EDX pulse processing and absorption issues (not in general, depends of matrix elements in specimen). Therefore a simple specimen tilt is often much better than an increase of excitation energy to improve count rate yield of an element of interest. If the count rate from specimen is sufficient, the use of a shorter pulse processor shaping-time increases the detection sensitivity (despite the worse energy resolution), because you can detect more counts in same time.
Only spectra simulation software is able to answer these complex excitation and absorption influences in advance, without any data acquisition for tests.
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Sorry if I digress a bit, but I am new to the field of EDX. I thought that higher voltages gave a higher signal in EDX, and so a higher sensitivity. Is it not true?
Not a lot of people know the answer to this, apparently Any advance on one reply??
Best wishes Chris
----- Original Message ----- X-from: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: {microscopy-at-microscopy.com} Sent: Monday, March 13, 2006 4:00 PM
Obviously photo-microscopy was going strong in 1904. There's no direct link to the article, so to find a rather incredible picture of 10 feet of bellows and plate camera connected to a simple brass compound microscope goto :
http://www.microscopy-uk.org.uk/
Click main resources ' Micscape article library'. Then click 'find' and enter '1904'
You will then get the link : 'Taking a photomicrograph .... in 1904 by Dave Walker, UK'
I think I may even be able to see a protein crystal on the microscope stage.
Any takers for an earlier example ?
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {c.jeffree-at-ed.ac.uk} To: {keith.morris-at-ucl.ac.uk} Sent: Wednesday, March 15, 2006 9:24 AM
Well, it's kind of a trick question. Would a photomicrograph of a protein containing structure showing birafrigence count? Would a X-ray defraction photo taken with a "micro" camera count? Ahh.. these are the question to settle over a beer or a nice cup of coffee.....
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c.jeffree-at-ed.ac.u k To: frank.karl-at-degussa.com cc: 03/15/2006 03:55 Subject: [Microscopy] Fw: First photomicrograph AM Please respond to c.jeffree
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Not a lot of people know the answer to this, apparently Any advance on one reply??
Best wishes Chris
----- Original Message ----- X-from: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: {microscopy-at-microscopy.com} Sent: Monday, March 13, 2006 4:00 PM
Barbara,
I have a set of manuals for the EM 300. Instructions for changing the filament are in the "Operating Instructions" volume -- there are several separate volumes -- Section D, pg 171 ff. Sounds like you don't have this volume. I can photocopy the pages and send them to you if you need. Or, if you or someone else using a Philips EM 300 needs the manuals, I can send them off -- we don't have one of these anymore. Note to the list: I also have a manual for an RCA EMU 4, if someone needs one. Might cost a pint or two at the next M&M meeting.
Phil
} This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both maloneyb-at-fiu.edu as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: maloneyb-at-fiu.edu } Name: Barbara } } Organization: FIU } } Title-Subject: [Filtered] How to change filament in Phillips 300 TEM } } Question: I have changed filaments in other instruments, but never } in this older model. The manual doesn't seem to cover this - does } anyone have the written procedure. Really would appreciate it } greatly. } Thanks } Barbara -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 24 -- From oshel1pe-at-cmich.edu Wed Mar 15 07:22:54 2006 4, 24 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FDMrQQ010632 4, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 07:22:53 -0600 4, 24 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 24 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k2FE154l025572; 4, 24 -- Wed, 15 Mar 2006 09:01:05 -0500 4, 24 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 24 -- Wed, 15 Mar 2006 08:22:39 -0500 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {f06230903c03dc4bd1d51-at-[141.209.160.132]} 4, 24 -- In-Reply-To: {200603150410.k2F4ATW6001828-at-ns.microscopy.com} 4, 24 -- References: {200603150410.k2F4ATW6001828-at-ns.microscopy.com} 4, 24 -- Date: Wed, 15 Mar 2006 08:22:48 -0500 4, 24 -- To: maloneyb-at-fiu.edu 4, 24 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 24 -- Subject: Re: [Microscopy] viaWWW: How to change filament in Phillips 300 4, 24 -- TEM 4, 24 -- Cc: Microscopy-at-microscopy.com 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-OriginalArrivalTime: 15 Mar 2006 13:22:39.0309 (UTC) FILETIME=[88405FD0:01C64833] 4, 24 -- X-CanItPRO-Stream: default 4, 24 -- X-Spam-Score: -4 () L_EXCH_MF 4, 24 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
It appears that John Dancer does have the honour of the first photomicrograph (of the flea), as suggested by Chris. However, ironically it is for inventing the procedure to create microphotographs on microscope slides (microfilm) that he is most remembered. I can't find any illustration of his 1840's 'gas powered microscope' on the web though, only pictures of him, his wife and many children.
A quick cut and paste biography of John Benjamin Dancer :
In 1840, when he was 28, John Dancer showed the world's first photomicrograph, of a Flea, in Liverpool, and he subsequently developed the microphotograph technique. In July 1840 he made a daguerreotype photograph of the flea, using a gas-illuminated microscope (this was a positive image on a metal support - the Daguerreotype was the first successful photographic process, the discovery being announced on 7 January 1839). In 1852 Dancer started making microscopic photographs - tiny photographs which could be viewed through a microscope. The microphotographs soon became popular and Dancer developed a large catalogue including photographs of the Royal family and Niagara Falls. Micro- photographs were then sold at one shilling (5p) each, or ten shillings and six pence (52.5p) for a dozen. He produced them commercially from about 1857. Although they sold poorly at first, within a few years they had become much sought after by science enthusiasts. He worked on various subjects, including landscapes, the Ten Commandments, and his most prestigious commission was for Queen Victoria, for whom he produced 5 miniature photographs of her family which were set in a signet ring - each picture being no more than 1/8th inch in diameter, and which were magnified in the ring by means of a jewel lens which he personally had cut. Dancer sold some 500 microphotograph slides, many of which were of well known paintings in art galleries. Particularly popular were slides of members of the Victorian Royal Family, of Emperor Napoleon, and of an 1858 20 banknote.
Although treated as a novelty until the 1920s, the microphotographic process eventually became 'microfilm'. Utilizing John Dancer's techniques, a French optician, Rene Dagron, was granted the first patent for microfilm in 1859 and began the first commercial microfilming enterprise. Dagron, during the Franco-Prussian War, demonstrated a practical use for microforms when carrier pigeons were used to transport microfilmed messages across German lines.
Otherwise Dancer's invention was not taken seriously, being variously described as "being of little or no practical use" and "childish and trivial". Yet, today, this invention is now used widely in banks, libraries and archives as a method of keeping important materials in an efficient, space-saving and economical way. He also invented the stereoscopic camera which he patented in 1853, contacts for electric alarms and a new form of illumination and photo-transparencies for use in lantern slide projection. Although not confirmed, it is widely believed Dancer created the first magic lantern photographic slide.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
"Fox Talbot was making images of botanical specimens with his solar microscope as early as February 1835, and is often credited as being the first photomicrographer. He especially loved photographing crystals but of what type is mostly unknown today."
Nancy
Nancy Smythe Department of Otolaryngology Head and Neck Surgery Medical University of South Carolina 843-792-8835 843-792-0368 Fax
==============================Original Headers============================== 6, 27 -- From smythen-at-musc.edu Wed Mar 15 08:16:37 2006 6, 27 -- Received: from caerbannog.musc.edu (caerbannog.musc.edu [128.23.203.43]) 6, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FEGaph024908 6, 27 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 08:16:36 -0600 6, 27 -- Received: from revere3.musc.edu (revere3.musc.edu [128.23.203.9]) 6, 27 -- by caerbannog.musc.edu (8.13.4/8.12.10) with ESMTP id k2FEEs7f008469 6, 27 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 09:14:54 -0500 6, 27 -- Received: from cl.musc.edu (cl.emr.musc.edu [128.23.151.3]) 6, 27 -- by revere3.musc.edu (8.12.9/8.12.9) with ESMTP id k2FEErAl017128 6, 27 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 09:14:53 -0500 (EST) 6, 27 -- Received: from CL-MTA by cl.musc.edu 6, 27 -- with Novell_GroupWise; Wed, 15 Mar 2006 09:14:31 -0500 6, 27 -- Message-Id: {s417daf7.004-at-cl.musc.edu} 6, 27 -- X-Mailer: Novell GroupWise Internet Agent 6.0.4 6, 27 -- Date: Wed, 15 Mar 2006 09:14:28 -0500 6, 27 -- From: "Nancy Smythe" {smythen-at-musc.edu} 6, 27 -- To: {microscopy-at-msa.microscopy.com} 6, 27 -- Subject: Re first photomicrograph 6, 27 -- Mime-Version: 1.0 6, 27 -- Content-Type: text/plain; charset=US-ASCII 6, 27 -- Content-Disposition: inline 6, 27 -- X-MUSC-MailScanner-Information: Please contact postmstr-at-musc.edu for more information 6, 27 -- X-MUSC-MailScanner: Found to be clean 6, 27 -- X-MUSC-MailScanner-SpamCheck: 6, 27 -- X-MailScanner-From: smythen-at-musc.edu 6, 27 -- Content-Transfer-Encoding: 8bit 6, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2FEGaph024908 ==============================End of - Headers==============================
} -----Original Message----- } From: biology-at-ucla.edu [mailto:biology-at-ucla.edu] } Sent: Tuesday, March 14, 2006 9:02 PM } To: Dusevich, Vladimir } Subject: [Microscopy] viaWWW: Cleaning Grids } } } } } -------------------------------------------------------------- } -------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } -------------- } } This Question/Comment was submitted to the Microscopy } Listserver using the WWW based Form at } http://www.microscopy.com/MLFormMail.html } -------------------------------------------------------------- } ------------- } Remember this posting is most likely not from a Subscriber, } so when replying } please copy both biology-at-ucla.edu as well as the } MIcroscopy Listserver } -------------------------------------------------------------- } ------------- } } Email: biology-at-ucla.edu } Name: Eric } } Organization: UCLA Medical Center } } Title-Subject: [Filtered] Cleaning Grids } } Question: This was never a problem till about a month ago.. } } Since about a month we have been having a problem keeping the } sections stuck to the grids. The grids are cleaned in 100% } Acetone and dried. Sections are picked up either from above } or below the water. } } When the grids are stained using the microwave staining } method we have been using for several years the sections come } off the grids... Every now and then the sections will stick } to the grids and everyhting is fine. } } Any suggestions about consistency? I have tried staining } less time in the microwave, but this does not make a difference... } } Any different ways to clean the grids so the sections will } stick better? } } } } -------------------------------------------------------------- } ------------- } } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Tue Mar 14 20:49:33 } 2006 12, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 12, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id k2F2nV3U009322 } 12, 12 -- for {microscopy-at-microscopy.com} ; Tue, 14 Mar } 2006 20:49:31 -0600 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p0611040ac03d311d0ae9-at-[206.69.208.22]} } 12, 12 -- Date: Tue, 14 Mar 2006 20:49:31 -0600 12, 12 -- To: } microscopy-at-microscopy.com 12, 12 -- From: biology-at-ucla.edu } (by way of MicroscopyListserver) 12, 12 -- Subject: viaWWW: } Cleaning Grids 12, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== }
==============================Original Headers============================== 6, 23 -- From DusevichV-at-umkc.edu Wed Mar 15 11:16:20 2006 6, 23 -- Received: from KC-MSXPROTO2.kc.umkc.edu (pop3.exchange.umkc.edu [134.193.143.155] (may be forged)) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FHGJE7019813 6, 23 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 11:16:20 -0600 6, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by KC-MSXPROTO2.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Wed, 15 Mar 2006 11:16:19 -0600 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: RE: [Microscopy] viaWWW: Cleaning Grids 6, 23 -- Date: Wed, 15 Mar 2006 11:16:18 -0600 6, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADBFC-at-KC-MSX1.kc.umkc.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [Microscopy] viaWWW: Cleaning Grids 6, 23 -- Thread-Index: AcZH3OYO5tPkLjyjS3SyB39oOjdAHAAdviOA 6, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 6, 23 -- To: {biology-at-ucla.edu} , {microscopy-at-msa.microscopy.com} 6, 23 -- X-OriginalArrivalTime: 15 Mar 2006 17:16:19.0395 (UTC) FILETIME=[2CDFB930:01C64854] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2FHGJE7019813 ==============================End of - Headers==============================
Using pre-embedding histochemistry, a client has infiltrated lung tissue with gold nanoparticles of various sizes. He would like to see the distribution of the particles on semithin sections and then examine the tissue with TEM. Does anybody out there have a good protocol for silver-intensifying the gold on 1 micron Epon sections for light microscopy? Any suggestions would be greatly appreciated.
Ralph Common Division of Human Pathology Michigan State University
==============================Original Headers============================== 3, 24 -- From rcommon-at-msu.edu Wed Mar 15 11:57:31 2006 3, 24 -- Received: from sys10.mail.msu.edu (sys10.mail.msu.edu [35.9.75.110]) 3, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FHvTK5027212 3, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 11:57:29 -0600 3, 24 -- Received: from [35.9.122.125] (helo=emlab) 3, 24 -- by sys10.mail.msu.edu with esmtpsa (Exim 4.52 #1) 3, 24 -- (TLSv1:RC4-MD5:128) 3, 24 -- id 1FJaFV-000205-4K 3, 24 -- for Microscopy-at-microscopy.com; Wed, 15 Mar 2006 12:57:29 -0500 3, 24 -- From: "Ralph Common" {rcommon-at-msu.edu} 3, 24 -- To: {Microscopy-at-microscopy.com} 3, 24 -- Subject: Nanogold in semithin sections 3, 24 -- Date: Wed, 15 Mar 2006 12:58:12 -0500 3, 24 -- Message-ID: {002f01c6485a$07a86210$7d7a0923-at-msu.edu} 3, 24 -- MIME-Version: 1.0 3, 24 -- Content-Type: text/plain; 3, 24 -- charset="iso-8859-1" 3, 24 -- Content-Transfer-Encoding: 7bit 3, 24 -- X-Priority: 3 (Normal) 3, 24 -- X-MSMail-Priority: Normal 3, 24 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) 3, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 3, 24 -- Importance: Normal 3, 24 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
Some brands of grids are routinely treated to prevent surface oxidation. If that treatment was not carried out properly on a particular batch of grids then sections and support films tend not to stick to the grids.
So long as its presence is not a problem for any reason you can dip the grids in a solution of Poly-L-Lysine which, when dry, helps the adhesion of sections. The Poly-L-Lysine is available from most EM supplies vendors.
Good luck
Ted
--- biology-at-ucla.edu wrote:
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==============================Original Headers============================== 10, 20 -- From drteddunne-at-yahoo.com Wed Mar 15 13:05:40 2006 10, 20 -- Received: from web33405.mail.mud.yahoo.com (web33405.mail.mud.yahoo.com [68.142.206.137]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2FJ5dDu007594 10, 20 -- for {microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 13:05:40 -0600 10, 20 -- Received: (qmail 21325 invoked by uid 60001); 15 Mar 2006 19:05:39 -0000 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 20 -- s=s1024; d=yahoo.com; 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 10, 20 -- b=QXIk89ojBUVCxHfMwJgmXomQT+Y47b0oP71D9CPlBsjufU0n8Yq6kF5JupBwUr7rh3YQepYAThkxF+WzP0fAeTi0nwJ0YJUCCt4wUs+Oe+Ca+WoL9EW1TDW3Qhzy4Ovvx473f2eeI3GKS3n48YYQlZwdmBh/XHad1gHeFAD0Xt8= ; 10, 20 -- Message-ID: {20060315190539.21323.qmail-at-web33405.mail.mud.yahoo.com} 10, 20 -- Received: from [202.47.247.116] by web33405.mail.mud.yahoo.com via HTTP; Wed, 15 Mar 2006 11:05:39 PST 10, 20 -- Date: Wed, 15 Mar 2006 11:05:39 -0800 (PST) 10, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 20 -- Subject: Re: [Microscopy] viaWWW: Cleaning Grids 10, 20 -- To: biology-at-ucla.edu 10, 20 -- Cc: microscopy-at-microscopy.com 10, 20 -- In-Reply-To: {200603150350.k2F3oXiX028013-at-ns.microscopy.com} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 10, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Hi Guys, I have a tantalum substrate covered with 30-40 nm tantalum oxide and I want to etch patterns through tantalum oxide to tantalum using HF. I am thinking to use EBL and PMMA as a photoresist, do you think PMMA stand against HF etching or the HF will etch thwe whole thing?
Thanks in advance
-- ********************************************************** Hany Ramadan Graduate student Chemistry department McMaster university, Hamilton, Ontario, Canada 905-525-9140 x: 26322 elsayeh-at-mcmaster.ca **********************************************************
==============================Original Headers============================== 4, 23 -- From ramadanhany-at-gmail.com Wed Mar 15 13:06:51 2006 4, 23 -- Received: from zproxy.gmail.com (zproxy.gmail.com [64.233.162.192]) 4, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FJ6oBo007819 4, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 13:06:50 -0600 4, 23 -- Received: by zproxy.gmail.com with SMTP id m22so195042nzf 4, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 11:06:50 -0800 (PST) 4, 23 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 23 -- s=beta; d=gmail.com; 4, 23 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 23 -- b=NHkumNmslSI2zJldfvRP8c2GKRkxSrY0kAWTX444AdD3MGrrkrsoQyVY2l+h+mVxsG0wAMLqJxrFRAOZkwnVIcT91TvoWR12EKJFdYDcOEOucEqh9GoAUDupNsQgrXawESptccR9mMjv/Y57xnvb0rFWOFTPipv8ziJOhU+Dpk0= 4, 23 -- Received: by 10.37.20.43 with SMTP id x43mr423587nzi; 4, 23 -- Wed, 15 Mar 2006 11:06:50 -0800 (PST) 4, 23 -- Received: by 10.37.12.3 with HTTP; Wed, 15 Mar 2006 11:06:50 -0800 (PST) 4, 23 -- Message-ID: {8d8ce5a30603151106v1b1f49a9x69073941151588a0-at-mail.gmail.com} 4, 23 -- Date: Wed, 15 Mar 2006 14:06:50 -0500 4, 23 -- From: "Hany Ramadan" {ramadanhany-at-gmail.com} 4, 23 -- To: Microscopy-at-microscopy.com 4, 23 -- Subject: Tantlum oxide etching aginst photoresist material 4, 23 -- MIME-Version: 1.0 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1 4, 23 -- Content-Disposition: inline 4, 23 -- Content-Transfer-Encoding: 8bit 4, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2FJ6oBo007819 ==============================End of - Headers==============================
By an interesting coincidence, the March issue of Microscopy Today has an article entitled "A Comparison of Photomicrographs Imaged Through a Late 18th C. Thomas Ribright, Cuff-Type, Brass Microscope and a Modern Olympus Optical Microscope", By D.Jones* and J.Reid.** *Retired Research Microbiologist Aberdeen and **School of Physics, The University, Aberdeen, Scotland. Apparently the microscope was manufactured in the late 1700's and was bought at auction recently. The microscope came in its original packing case and was accompanied by a box of accessories, which included some micro-slides by Dancer, ca 1850-60 as described in Morris' post. Jones and Reid took micrographs for the article with an Olympus OM10 single reflex 35mm camera, with lens removed and two extension tubes (20mm and 12mm) attached, fitted to the eyepiece of the microscope and held in place with a tripod stand; the source of light was a 60 watt tungsten electric light bulb in an angle-poise lamp stand. Fujichrome Professional 64T color film was used to record images. The MT article contains images of two Dancer slides, one of a very youthful Queen Victoria and the Prince Regent and another one of Trafalgar Square. No dates are given for the Dancer slides. The authors give the following reference, which may be of interest to anyone interested in Dancer photomicrographs, etc.: "Bracegirdle, B. and McCormick, J. B. The Microscopic Photographs of J.B.Dancer. Science Heritage Limited, Chicago, Illinois, 1993."
Ron Anderson, Editor Microscopy Today
keith.morris-at-ucl.ac.uk wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } It appears that John Dancer does have the honour of the first } photomicrograph (of the flea), as suggested by Chris. However, ironically it } is for inventing the procedure to create microphotographs on microscope } slides (microfilm) that he is most remembered. I can't find any illustration } of his 1840's 'gas powered microscope' on the web though, only pictures of } him, his wife and many children. } } A quick cut and paste biography of John Benjamin Dancer : } } In 1840, when he was 28, John Dancer showed the world's first } photomicrograph, of a Flea, in Liverpool, and he subsequently developed the } microphotograph technique. In July 1840 he made a daguerreotype photograph } of the flea, using a gas-illuminated microscope (this was a positive image } on a metal support - the Daguerreotype was the first successful photographic } process, the discovery being announced on 7 January 1839). In 1852 Dancer } started making microscopic photographs - tiny photographs which could be } viewed through a microscope. The microphotographs soon became popular and } Dancer developed a large catalogue including photographs of the Royal family } and Niagara Falls. Micro- photographs were then sold at one shilling (5p) } each, or ten shillings and six pence (52.5p) for a dozen. He produced them } commercially from about 1857. Although they sold poorly at first, within a } few years they had become much sought after by science enthusiasts. He } worked on various subjects, including landscapes, the Ten Commandments, and } his most prestigious commission was for Queen Victoria, for whom he produced } 5 miniature photographs of her family which were set in a signet ring - each } picture being no more than 1/8th inch in diameter, and which were magnified } in the ring by means of a jewel lens which he personally had cut. Dancer } sold some 500 microphotograph slides, many of which were of well known } paintings in art galleries. Particularly popular were slides of members of } the Victorian Royal Family, of Emperor Napoleon, and of an 1858 20 banknote. } } Although treated as a novelty until the 1920s, the microphotographic process } eventually became 'microfilm'. Utilizing John Dancer's techniques, a French } optician, Rene Dagron, was granted the first patent for microfilm in 1859 } and began the first commercial microfilming enterprise. Dagron, during the } Franco-Prussian War, demonstrated a practical use for microforms when } carrier pigeons were used to transport microfilmed messages across German } lines. } } Otherwise Dancer's invention was not taken seriously, being variously } described as "being of little or no practical use" and "childish and } trivial". Yet, today, this invention is now used widely in banks, libraries } and archives as a method of keeping important materials in an efficient, } space-saving and economical way. He also invented the stereoscopic camera } which he patented in 1853, contacts for electric alarms and a new form of } illumination and photo-transparencies for use in lantern slide projection. } Although not confirmed, it is widely believed Dancer created the first magic } lantern photographic slide. } } Keith } } ---------------------------------------------------------- } Dr Keith J Morris } Imaging Facilities Manager } Cell Biology Division } Institute of Ophthalmology } University College London } 11-43 Bath Street } London EC1V 9EL } } Tel: 020 7608 4050 } Fax: 020 7608 4034 } email: keith.morris-at-ucl.ac.uk } } } ==============================Original Headers============================== } 9, 27 -- From keith.morris-at-ucl.ac.uk Wed Mar 15 08:04:56 2006 } 9, 27 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) } 9, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FE4qu7021334 } 9, 27 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 08:04:54 -0600 } 9, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) } 9, 27 -- by vscani-d.ucl.ac.uk with smtp (Exim 4.51) } 9, 27 -- id 1FJWcM-0005uL-7N } 9, 27 -- for Microscopy-at-microscopy.com; Wed, 15 Mar 2006 14:04:50 +0000 } 9, 27 -- Message-ID: {00be01c64839$5109a480$7b865290-at-keithhigrade} } 9, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} } 9, 27 -- To: {Microscopy-at-microscopy.com} } 9, 27 -- Subject: [Microscopy] Fw: First photomicrograph } 9, 27 -- Date: Wed, 15 Mar 2006 14:04:03 -0000 } 9, 27 -- MIME-Version: 1.0 } 9, 27 -- Content-Type: text/plain; } 9, 27 -- format=flowed; } 9, 27 -- charset="iso-8859-1"; } 9, 27 -- reply-type=original } 9, 27 -- Content-Transfer-Encoding: 7bit } 9, 27 -- X-Priority: 3 } 9, 27 -- X-MSMail-Priority: Normal } 9, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 } 9, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 } 9, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information } 9, 27 -- X-UCL-MailScanner: Found to be clean } 9, 27 -- X-UCL-MailScanner-SpamCheck: } 9, 27 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 19 -- From microscopytoday-at-tampabay.rr.com Wed Mar 15 15:40:44 2006 5, 19 -- Received: from ms-smtp-02.tampabay.rr.com (ms-smtp-02-smtplb.tampabay.rr.com [65.32.5.132]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2FLehGL029059 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Wed, 15 Mar 2006 15:40:43 -0600 5, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 19 -- by ms-smtp-02.tampabay.rr.com (8.13.4/8.13.4) with ESMTP id k2FLebi2014810; 5, 19 -- Wed, 15 Mar 2006 16:40:42 -0500 (EST) 5, 19 -- Message-ID: {441889D3.5070106-at-tampabay.rr.com} 5, 19 -- Date: Wed, 15 Mar 2006 16:40:35 -0500 5, 19 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: keith.morris-at-ucl.ac.uk, Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] Fw: First photomicrograph 5, 19 -- References: {200603151407.k2FE7CX1022076-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200603151407.k2FE7CX1022076-at-ns.microscopy.com} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Hi all, I just had a request to use our TEM to look at the protozoan Toxoplasma. The samples are unfixed - just dried onto formvar-coated grids. Is this a safe procedure? I'm use to all samples being fixed.
Any advice would be greatly appreciated. best, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Enhancement for LM has been done for decades, both with published recipes (Danscher, Burry, Bienz and Springall and Lackie from the Hammersmith EM research group in London) as well as with commercial reagents. There are a number of companies who produce silver (and gold) enhancement reagents for light and for electron microscopy. If the particles are sufficiently enhanced you will be able to pick up individual particles in the light microscope bright field image, and certainly in epi-polarisation mode. In our experience enhancement will mostly be limited to particles on or close to the surface: they have to be exposed to become enhanced. Depending on the resin there seems to be some penetration, however, with larger particles on the surface, smaller ones below the surface. To visualise the particles in LM the particles need to be relatively big, and visualising the same specimen in EM might not be ideal. But I guess your client wants to check the specimens in LM and if a signal is found, look at unenhanced ones in EM? I initially (probably mistakenly) assumed the study was about discriminating between particle sizes after enhancement. Even though the size of the enhanced particles will somewhat depend on the initial size of the gold particles, I seriously doubt it would be possible to discriminate between sizes using LM techniques. In fact that may even be hard in electron microscopy, unless the initial particles were significantly different in size. On the other hand, double labelling using silver enhancement and ultra small gold particles has been successfully done in pre-embedding EM (Yi, H., J. L.M. Leunissen, G.-M. Shi, C.-A. Gutekunst, and S. M. Hersch. A Novel Procedure for Pre-embedding Double Immunogold-Silver Labeling at the Ultrastructural Level; J. Histochem. Cytochem., March 1, 2001; 49(3): 279 - 284)
Should you require more info, please feel welcome to contact me off- list.
Jan Leunissen
On 16/03/2006, at 7:01 AM, rcommon-at-msu.edu wrote: } } Using pre-embedding histochemistry, a client has infiltrated lung } tissue } with gold nanoparticles of various sizes. He would like to see the } distribution of the particles on semithin sections and then examine } the } tissue with TEM. Does anybody out there have a good protocol for } silver-intensifying the gold on 1 micron Epon sections for light } microscopy? } Any suggestions would be greatly appreciated. } } Ralph Common } Division of Human Pathology } Michigan State University
Aurion - President Present Address: Costerweg 5 EM-Unit 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4797109 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://ocem.otago.ac.nz ------------------------------------------------------------------------ --------
==============================Original Headers============================== 12, 23 -- From leunissen-at-aurion.nl Wed Mar 15 19:18:43 2006 12, 23 -- Received: from mta201-rme.xtra.co.nz (mta201-rme.xtra.co.nz [210.86.15.144]) 12, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2G1IfkU018685 12, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 19:18:42 -0600 12, 23 -- Received: from mta2-rme.xtra.co.nz ([210.86.15.192]) 12, 23 -- by mta201-rme.xtra.co.nz with ESMTP 12, 23 -- id {20060316011839.QSSP29457.mta201-rme.xtra.co.nz-at-mta2-rme.xtra.co.nz} 12, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Mar 2006 14:18:39 +1300 12, 23 -- Received: from [192.168.1.20] ([222.153.174.203]) by mta2-rme.xtra.co.nz 12, 23 -- with ESMTP 12, 23 -- id {20060316011839.SKM18564.mta2-rme.xtra.co.nz-at-[192.168.1.20]} 12, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Mar 2006 14:18:39 +1300 12, 23 -- Mime-Version: 1.0 (Apple Message framework v746.2) 12, 23 -- In-Reply-To: {200603151801.k2FI1QUS028241-at-ns.microscopy.com} 12, 23 -- References: {200603151801.k2FI1QUS028241-at-ns.microscopy.com} 12, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 23 -- Message-Id: {FE407FAA-B7FD-47F4-947A-E6FE60EF3B0C-at-aurion.nl} 12, 23 -- Content-Transfer-Encoding: 7bit 12, 23 -- From: Jan Leunissen {leunissen-at-aurion.nl} 12, 23 -- Subject: Re: [Microscopy] Nanogold in semithin sections 12, 23 -- Date: Thu, 16 Mar 2006 14:18:38 +1300 12, 23 -- To: Microscopy-at-microscopy.com 12, 23 -- X-Mailer: Apple Mail (2.746.2) ==============================End of - Headers==============================
On Mar 14, 2006, at 6:50 PM, pekysar-at-ucdavis.edu wrote:
} Title-Subject: [Filtered] Low Dose TEM } } Question: Hello all, } We have a client who is trying to use the low dose function of our FEI } CM120. He is looking at magnetic nano particles on a substrate. He was } able to make it work but is still getting damage to his sample. } Unfortuately, neither of the techs here have used the low dose so we } are quite unfamiliar with it and aren't much help. } He would like to know what radius he should be using and we would like } to hear from anyone who has experience with the low dose on this tool } (or any tips, for that matter) which might help him achieve success. } We would all appreciate any help! } Cheers, } Pat Kysar } University of California, Davis } Medical School, Pathology } EM Lab } Dear Pat, First, you need to be sure that you are using a pre-specimen shutter, and that it is open only during the time the image is being recorded and not when the CCD read-out is happening. This will minimize dose to the specimen. Second, when setting up the LowDose parameters, set the focus offset so that there is no overlap of the beam in focus state with the area of the CCD image in exposure state--this will, of course, vary with the magnifications in the two states. Note that this also requires that the beam size in the focus state is only a little larger than the size of the image in that state. This is quite simple to achieve on the Tecnai T12 (which we have), but may be more difficult on the CM120 (with which I have no experience). Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Mar 15 19:36:29 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2G1aSLc021949 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 19:36:29 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP 4, 22 -- id 2461A3621F; Wed, 15 Mar 2006 17:36:28 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP 4, 22 -- id 41F3810A892; Wed, 15 Mar 2006 17:36:27 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603150250.k2F2oMSS009582-at-ns.microscopy.com} 4, 22 -- References: {200603150250.k2F2oMSS009582-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {c07ee33431c96f1f27b4a91aaf49803e-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: Low Dose TEM 4, 22 -- Date: Wed, 15 Mar 2006 17:45:12 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com, pekysar-at-ucdavis.edu 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both scott.streiker-at-udri.udayton.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: scott.streiker-at-udri.udayton.edu Name: Scott Streiker
Organization: University of Dayton Research Institute
Title-Subject: [Filtered] Protocol for TEM of Polymer Materials
Question: References, resources, protocols sought for preparation of polymer samples for TEM. Thanks
Scott Streiker wrote: ===================================================== Title-Subject: [Filtered] Protocol for TEM of Polymer Materials
Question: References, resources, protocols sought for preparation of polymer samples for TEM. ====================================================== We have found the text Polymer Microscopy by Linda C. Sawyer and David T. Grubb, ISBN: 0412257106, Springer, to be about as close to being a "bible" as one can get, especially if one is studying polymer blends and rubber modified systems. First published in 1987, and updated in 1995, the content seems to be nearly as timely today as the day it was published. I think the book is out of print but as of today, Amazon seems to have availability both new and used.
If you are looking to visualize lamellar structures in crystalline polymers, and/or the deformation of crystalline polymers, the "bible" for that part of polymer microscopy is Polymer Single Crystals by Philip H. Geil, published in 1962. No that is not a typo for the date. For that end of polymer microscopy, the book is nearly as relavent today as it was then. It is out of print and not listed on Amazon but it probably would be found in most university libraries.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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==============================Original Headers============================== 12, 29 -- From cgarber-at-2spi.com Wed Mar 15 23:41:32 2006 12, 29 -- Received: from s-utl02-atpop.stsn.net (s-utl02-atpop.stsn.net [72.254.128.202]) 12, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2G5fVja017817 12, 29 -- for {microscopy-at-msa.microscopy.com} ; Wed, 15 Mar 2006 23:41:31 -0600 12, 29 -- Received: from s-utl02-atpop.stsn.net ([127.0.0.1]) 12, 29 -- by s-utl02-atpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006031600413131337 12, 29 -- ; Thu, 16 Mar 2006 00:41:31 -0500 12, 29 -- X-Spam-Status: No, hits=0.0 required=9.9 12, 29 -- tests=ALL_TRUSTED: -2.4,AWL: 0.119,SARE_RECV_ADDR: 0.027 12, 29 -- X-Spam-Level: 12, 29 -- Received: from ibm1x23g2abfyg ([10.0.91.139]) 12, 29 -- by s-utl02-atpop.stsn.net; 12, 29 -- Thu, 16 Mar 2006 00:41:29 -0500 12, 29 -- Message-ID: {001201c648bc$43d80580$8b5b000a-at-ibm1x23g2abfyg} 12, 29 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 12, 29 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 12, 29 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 29 -- Cc: {scott.streiker-at-udri.udayton.edu} 12, 29 -- Subject: References for TEM of polymers 12, 29 -- Date: Thu, 16 Mar 2006 00:41:15 -0500 12, 29 -- MIME-Version: 1.0 12, 29 -- Content-Type: text/plain; 12, 29 -- charset="Windows-1252" 12, 29 -- X-Priority: 3 12, 29 -- X-MSMail-Priority: Normal 12, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 12, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 12, 29 -- Content-Transfer-Encoding: 8bit 12, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2G5fVja017817 ==============================End of - Headers==============================
I face 3 problems and I wondered if you could not solve them by the same solution, namely carbon-coating OVER the sections.
My first problem is with semi-thick sections (for tomography): they don't stick to the grids during contrasting and I loose them! I thought that perhaps carbon-coating after the grids are deposited on the grid would help keeping them on the grid without disturbing contrasting ?
My second problem is with ultra-thin sections: when I do EDX analysis (at 200 keV) on 70 nm thick sections on formvar film, they suffer much from the beam and usually I don't see anything when I pass in STEM mode because the area has been vaporized ;-) I thought that perhaps carbon-coating the contrasted sections would help disperse the energy of the beam?
My third problem deals with 50 nm sections deposited on grid without formvar, which are very unstable under 80 keV. Well I have difficulties to make formvar films which stick to the grids, they tend to disappear in the contrasting solutions. I wondered if I could not deposit 50 nm thick sections on grids without formvar and then carbon-coating them (so over the sections).
P.S: I clean the grids by sonicating in acetone.
Thanks in advance for your humble opinions.
Stephane (without "i")
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 9, 18 -- From nizets2-at-yahoo.com Thu Mar 16 02:35:21 2006 9, 18 -- Received: from web37406.mail.mud.yahoo.com (web37406.mail.mud.yahoo.com [209.191.87.59]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2G8ZKvR029350 9, 18 -- for {microscopy-at-microscopy.com} ; Thu, 16 Mar 2006 02:35:20 -0600 9, 18 -- Received: (qmail 57273 invoked by uid 60001); 16 Mar 2006 08:35:20 -0000 9, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 18 -- s=s1024; d=yahoo.com; 9, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 18 -- b=CzFyOchbrHvDMLaexVt0ODAulB0nHLAicLGtaB2N5z/7ZfDs5WSrFRkOb9/RGDpsjsqJuZ5hcuQ3miYi2ppHnF7wm1oOezskOh/G6hOCIAxlXOYfX3hBwAEHacVfrbvvd1pnakzYUSW3/GEZ6l5T5zdu9VmAf36skYqowyjpObI= ; 9, 18 -- Message-ID: {20060316083520.57271.qmail-at-web37406.mail.mud.yahoo.com} 9, 18 -- Received: from [80.122.101.102] by web37406.mail.mud.yahoo.com via HTTP; Thu, 16 Mar 2006 00:35:20 PST 9, 18 -- Date: Thu, 16 Mar 2006 00:35:20 -0800 (PST) 9, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 18 -- Subject: carbon post-coating ;-) in TEM 9, 18 -- To: microscopy-at-microscopy.com 9, 18 -- MIME-Version: 1.0 9, 18 -- Content-Type: text/plain; charset=iso-8859-1 9, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Here is an article that ought to be of use to you:
Bassett, D. C., Olley, R. H. and Vaughan, A. S. (2003) Specimen Preparation for TEM of Polymers, in Pethrick, R. A. and Viney, C., Eds. Techniques in Polymer Organisation and Morphology Characterisation, chapter 3, pp. 73-110. Wiley.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
} From: scott.streiker-at-udri.udayton.edu } Reply-To: scott.streiker-at-udri.udayton.edu } To: hinmeigeng-at-hotmail.com } Subject: [Microscopy] viaWWW: Protocol for TEM of Polymer Materials } Date: Wed, 15 Mar 2006 23:00:24 -0600 } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 12, 22 -- From hinmeigeng-at-hotmail.com Thu Mar 16 02:41:15 2006 12, 22 -- Received: from hotmail.com (bay101-f32.bay101.hotmail.com [64.4.56.42]) 12, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2G8fEEE030528 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 02:41:15 -0600 12, 22 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 12, 22 -- Thu, 16 Mar 2006 00:41:14 -0800 12, 22 -- Message-ID: {BAY101-F321721DCF1014E3F03241ACAE70-at-phx.gbl} 12, 22 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 12, 22 -- Thu, 16 Mar 2006 08:41:11 GMT 12, 22 -- X-Originating-IP: [86.128.212.193] 12, 22 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 12, 22 -- X-Sender: hinmeigeng-at-hotmail.com 12, 22 -- Reply-To: R.H.Olley-at-reading.ac.uk 12, 22 -- In-Reply-To: {200603160500.k2G50OsZ010327-at-ns.microscopy.com} 12, 22 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 12, 22 -- To: scott.streiker-at-udri.udayton.edu, Microscopy-at-MSA.Microscopy.Com 12, 22 -- Cc: R.H.Olley-at-reading.ac.uk 12, 22 -- Subject: RE: [Microscopy] viaWWW: Protocol for TEM of Polymer Materials 12, 22 -- Date: Thu, 16 Mar 2006 08:41:11 +0000 12, 22 -- Mime-Version: 1.0 12, 22 -- Content-Type: text/plain; format=flowed 12, 22 -- X-OriginalArrivalTime: 16 Mar 2006 08:41:14.0275 (UTC) FILETIME=[62666730:01C648D5] ==============================End of - Headers==============================
I have done many Toxo TEMs. Drying the parasite kills it. There is no possibility of spoors or other environmentally stable forms. Unless you have a high power TEM, there will not be too much to see. Depends on what your user wants. David
On Mar 15, 2006, at 4:30 PM, beth-at-plantbio.uga.edu wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Hi all, } I just had a request to use our TEM to look at the protozoan } Toxoplasma. The samples are unfixed - just dried onto formvar-coated } grids. } Is this a safe procedure? I'm use to all samples being fixed. } } Any advice would be greatly appreciated. } best, } Beth } } } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } http://www.plantbio.uga.edu/emlab } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } *********************************************************************** } * } *** } } } } ==============================Original } Headers============================== } 10, 18 -- From beth-at-plantbio.uga.edu Wed Mar 15 17:00:44 2006 } 10, 18 -- Received: from dogwood.plantbio.uga.edu } (dogwood.plantbio.uga.edu [128.192.26.2]) } 10, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k2FN0iSY007206 } 10, 18 -- for {microscopy-at-microscopy.com} ; Wed, 15 Mar 2006 17:00:44 } -0600 } 10, 18 -- Received: from plantbio.uga.edu ([128.192.26.46]) } 10, 18 -- by dogwood.plantbio.uga.edu } 10, 18 -- (using TLSv1/SSLv3 with cipher DES-CBC3-SHA (168 bits)) } 10, 18 -- for microscopy-at-microscopy.com; } 10, 18 -- Wed, 15 Mar 2006 18:00:38 -0500 } 10, 18 -- Date: Wed, 15 Mar 2006 18:00:41 -0500 } 10, 18 -- Mime-Version: 1.0 (Apple Message framework v553) } 10, 18 -- Content-Type: text/plain; delsp=yes; charset=US-ASCII; } format=flowed } 10, 18 -- Subject: tem of toxoplasma } 10, 18 -- From: Beth Richardson {beth-at-plantbio.uga.edu} } 10, 18 -- To: microscopy microscopy {microscopy-at-microscopy.com} } 10, 18 -- Content-Transfer-Encoding: 7bit } 10, 18 -- Message-Id: } {8698B2A3-B477-11DA-830F-000393137C00-at-plantbio.uga.edu} } 10, 18 -- X-Mailer: Apple Mail (2.553) } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-Arizona.edu Thu Mar 16 09:40:38 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2GFeblW029764 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 09:40:37 -0600 5, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id F1E67D6F13D 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 08:40:36 -0700 (MST) 5, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id B80CAD708DB 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Mar 2006 08:40:35 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 5, 22 -- In-Reply-To: {200603152330.k2FNUjqm013179-at-ns.microscopy.com} 5, 22 -- References: {200603152330.k2FNUjqm013179-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {48b870b177732234386709311ac3c686-at-Arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-Arizona.edu} 5, 22 -- Subject: Re: [Microscopy] tem of toxoplasma 5, 22 -- Date: Thu, 16 Mar 2006 08:40:35 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.623) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
On Mar 16, 2006, at 12:36 AM, nizets2-at-yahoo.com wrote:
} I face 3 problems and I wondered if you could not } solve them by the same solution, namely carbon-coating } OVER the sections. } } My first problem is with semi-thick sections (for } tomography): they don't stick to the grids during } contrasting and I loose them! I thought that perhaps } carbon-coating after the grids are deposited on the } grid would help keeping them on the grid without } disturbing contrasting ? } } My second problem is with ultra-thin sections: when I } do EDX analysis (at 200 keV) on 70 nm thick sections } on formvar film, they suffer much from the beam and } usually I don't see anything when I pass in STEM mode } because the area has been vaporized ;-) I thought that } perhaps carbon-coating the contrasted sections would } help disperse the energy of the beam? } } My third problem deals with 50 nm sections deposited } on grid without formvar, which are very unstable under } 80 keV. Well I have difficulties to make formvar films } which stick to the grids, they tend to disappear in } the contrasting solutions. I wondered if I could not } deposit 50 nm thick sections on grids without formvar } and then carbon-coating them (so over the sections). } } P.S: I clean the grids by sonicating in acetone. } } Thanks in advance for your humble opinions. } Dear Stephane, Carbon coating might not affect your semi-thick sections coming loose during contrasting, but a previous post suggested coating the grid with polylysine before placing the sections on it, and another suggested a brief acid dip to roughen the grid surface, so I'd try these. Carbon coating semi-thick specimens, however, will aid both thermal and electrical conductivity, so I do recommend that you coat your specimens, and if you are using a formvar substrate, carbon coat that before placing the sections. From this answer, you can surmise that my answer to your second problem is to carbon coat both the formvar before placing the thin sections and carbon coat the sections after. I would suggest either acid-dipping the grids and covering them with formvar, or using a higher-mesh grid without formvar (depending on how large an unobstructed field of view you need) and carbon coat the sections to increase stability of your 50 nm sections. In the latter case I even suggest coating both sides with carbon--being careful not to dislodge the sections when you coat the underside of the grid, of course. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Thu Mar 16 13:06:44 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2GJ6hDo009886 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 16 Mar 2006 13:06:43 -0600 4, 22 -- Received: from localhost (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 2EDE810AA79 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 16 Mar 2006 11:06:43 -0800 (PST) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 08FD710AA62 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 16 Mar 2006 11:06:42 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200603160836.k2G8aGps029523-at-ns.microscopy.com} 4, 22 -- References: {200603160836.k2G8aGps029523-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {64e50971d37899b5d5bebd33aa3d09da-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] carbon post-coating ;-) in TEM 4, 22 -- Date: Thu, 16 Mar 2006 11:15:32 -0800 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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Email: scott.streiker-at-udri.udayton.edu Name: Scott Streiker
Organization: University of Dayton reserach Institute
Title-Subject: [Filtered] Polymer preparation for TEM
Question: Thanks to all who responded to question re: preparation polymer samples for TEM.
As a result of good advice I have acquired copy "Polymer Microscopy by, Sawyer and Grubb Cheers,
Scott Streiker Electron Microscopist NEST Laboratory University of Dayton Research Institute Scott.Streiker-at-udri.udayton.edu Phone: (937)229-5776
Browsing through this month's European Semiconductor this morning I find an article on page 46 beginning..
"Hillsboro (Oregon, USA) based FEI has been in the microscopy business since 1949, when it produced the world's first Transmission Electron Microscope (TEM). Today FEI manufactures a full range of microscopes..."
Now I'm no history buff, but I didn't see an FEI label on the replica of the first TEM in the Science museum in London when I visited last year.. How can they make this claim?
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Fri Mar 17 05:48:23 2006 8, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2HBmM8R011251 8, 31 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 05:48:22 -0600 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-5.tower-78.messagelabs.com!1142596059!44508703!15 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 13263 invoked from network); 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-5.tower-78.messagelabs.com with SMTP; 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 8, 31 -- Fri, 17 Mar 2006 11:48:15 +0000 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 31 -- Content-class: urn:content-classes:message 8, 31 -- MIME-Version: 1.0 8, 31 -- Content-Type: text/plain; 8, 31 -- charset="us-ascii" 8, 31 -- Subject: The first TEM 8, 31 -- Date: Fri, 17 Mar 2006 11:48:14 -0000 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047C36-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: The first TEM 8, 31 -- Thread-Index: AcZJuKzBYGI+b8m5RBapPz+el04xIw== 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 17 Mar 2006 11:48:15.0713 (UTC) FILETIME=[AD4FB510:01C649B8] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2HBmM8R011251 ==============================End of - Headers==============================
Who's first? While some might argue that it was JJ Thompson with his work on cathode ray, let us skip forward to Ernst Ruska and Max Knoll who developed the first magnetic lens electron microscope and published in 1932 (Ann. d. Physik 12, 607 (1932). In North America, J Hiller and A Prebus at the University of Toronto, built and operated the first TEM. I will never forget reading about how they dissolved rubber bands in solvent to produce vacuum greases. (I just have to squeeze a tube...) Hiller left to join RCA to manufacture the first commercial TEM in North America, the EMB. I don't have a exact date, but Hillier and Ramberg published an article about this in J. Appl. Phys., 18, 48, 1947. Meanwhile in Europe, von Borries and Ruska were developing/manufacturing an instrument similar to the EMB at Siemens-Halske and published in Z. wiss Mikroskop 56, 317 (1939).
When I started at Goodyear tire, they had an RCA EMU-3 that was just about 25 years old, placing it's purchase in 1954 and was only a few years younger then I was. I have a publication containing a line drawing of the lab's founder, Wilisfort (I'm sure that a mis-spelling) using a EBM during the second World War. I had no idea I was walk in the shadow of such illuminated history.
The question to answer is what companies evolved into FEI? If it wasn't either RCA or Siemens-Halske the claim should be rejected.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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richard.beanland-at- bookham.com To: frank.karl-at-degussa.com cc: 03/17/2006 07:02 Subject: [Microscopy] The first TEM AM Please respond to richard.beanland
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Browsing through this month's European Semiconductor this morning I find an article on page 46 beginning..
"Hillsboro (Oregon, USA) based FEI has been in the microscopy business since 1949, when it produced the world's first Transmission Electron Microscope (TEM). Today FEI manufactures a full range of microscopes..."
Now I'm no history buff, but I didn't see an FEI label on the replica of the first TEM in the Science museum in London when I visited last year.. How can they make this claim?
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Fri Mar 17 05:48:23 2006 8, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2HBmM8R011251 8, 31 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 05:48:22 -0600 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-5.tower-78.messagelabs.com!1142596059!44508703!15 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 13263 invoked from network); 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-5.tower-78.messagelabs.com with SMTP; 17 Mar 2006 11:48:19 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 8, 31 -- Fri, 17 Mar 2006 11:48:15 +0000 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 31 -- Content-class: urn:content-classes:message 8, 31 -- MIME-Version: 1.0 8, 31 -- Content-Type: text/plain; 8, 31 -- charset="us-ascii" 8, 31 -- Subject: The first TEM 8, 31 -- Date: Fri, 17 Mar 2006 11:48:14 -0000 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047C36-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: The first TEM 8, 31 -- Thread-Index: AcZJuKzBYGI+b8m5RBapPz+el04xIw== 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 17 Mar 2006 11:48:15.0713 (UTC) FILETIME=[AD4FB510:01C649B8] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2HBmM8R011251 ==============================End of - Headers==============================
==============================Original Headers============================== 35, 18 -- From frank.karl-at-degussa.com Fri Mar 17 07:26:40 2006 35, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 35, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HDQdUk021737 35, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 17 Mar 2006 07:26:39 -0600 35, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 35, 18 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2HDQX9L030681; 35, 18 -- Fri, 17 Mar 2006 14:26:34 +0100 35, 18 -- In-Reply-To: {200603171202.k2HC2YWs013693-at-ns.microscopy.com} 35, 18 -- Subject: Re: [Microscopy] The first TEM 35, 18 -- To: richard.beanland-at-bookham.com, microscopy-at-msa.microscopy.com 35, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 35, 18 -- Message-ID: {OF7447929D.9CF0BD6C-ON85257134.00470EE7-85257134.0049D191-at-degussa.com} 35, 18 -- From: frank.karl-at-degussa.com 35, 18 -- Date: Fri, 17 Mar 2006 08:26:17 -0500 35, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 35, 18 -- 03/17/2006 07:26:35 AM 35, 18 -- MIME-Version: 1.0 35, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Quite easily. But I think they will find if they look into their innermost souls (if I am not spawning some appalling oxymoron here) that FEI was not even a gleam in it's creator's eye in 1949. Indeed, probably FEI's creators were not even yet gleams in their parents eyes. In fact, ...... no, forget it!
*Philips* introduced a commercial TEM in 1949, but Zeiss also claim a first commercial TEM (EM7) in 1949 and JEOL's first was in 1950, but Leeds University web site reports receiving its first TEM on 6 January 1944, installed in the ladies lavatory of the Textiles laboratory! Is it not the case that the first commercial TEMs were designed by Ruska and produced by Siemens, and that several tens of instruments were out there by 1945?? Ruska and Knoll built a TEM in 1931, and Ruska later built one by himself in 1933 that bettered the resolution of light microscopes.
see http://nobelprize.org/physics/laureates/1986/ruska-autobio.html
Chris
----- Original Message ----- X-from: {richard.beanland-at-bookham.com} To: {cjeffree-at-staffmail.ed.ac.uk} Sent: Friday, March 17, 2006 11:51 AM
How can FEI claim to have "produced the world's first Transmission Electron Microscope (TEM)"?
Blowed if I know. From my understanding, it's not the first TEM (1931, Ruska and Knoll), or the first commercial TEM (1936, MetropolitanVickers EM1), or even the first commercial series of TEMs (1939, Siemens). If these don't count as TEMs in some way, I'd like to know.
I think some PR person has read a line saying "Philips first TEM" and re-written it as "The first TEM"...
Mike Fay School of Mechanical, Materials and Manufacturing Engineering Nottingham University University Park Nottingham NG7 2RD tel 0115 8466081
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==============================Original Headers============================== 10, 24 -- From michael.fay-at-nottingham.ac.uk Fri Mar 17 07:41:18 2006 10, 24 -- Received: from mozart.is.nottingham.ac.uk (mozart.is.nottingham.ac.uk [128.243.40.93]) 10, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HDfHuG025454 10, 24 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 07:41:17 -0600 10, 24 -- Received: from ccw0m1.ccc.nottingham.ac.uk ([128.243.220.65] helo=ccw0m1.nottingham.ac.uk) 10, 24 -- by mozart.is.nottingham.ac.uk with esmtp (Exim 3.36 #2) 10, 24 -- id 1FKFCN-0000rY-00 10, 24 -- for microscopy-at-microscopy.com; Fri, 17 Mar 2006 13:40:59 +0000 10, 24 -- Received: from Gwweb1-MTA by ccw0m1.nottingham.ac.uk 10, 24 -- with Novell_GroupWise; Fri, 17 Mar 2006 13:41:01 +0000 10, 24 -- Message-Id: {s41abc6d.056-at-ccw0m1.nottingham.ac.uk} 10, 24 -- X-Mailer: Novell GroupWise Internet Agent 6.0.3 10, 24 -- Date: Fri, 17 Mar 2006 13:40:32 +0000 10, 24 -- From: "Michael Fay" {Michael.Fay-at-nottingham.ac.uk} 10, 24 -- To: {microscopy-at-microscopy.com} 10, 24 -- Subject: Re: [Microscopy] The first TEM 10, 24 -- Mime-Version: 1.0 10, 24 -- Content-Type: text/plain; charset=US-ASCII 10, 24 -- Content-Disposition: inline 10, 24 -- X-MailScanner-Information: Please contact staff-it-helpline-at-nottingham.ac.uk for more information 10, 24 -- X-MailScanner: Found to be clean 10, 24 -- X-MailScanner-From: michael.fay-at-nottingham.ac.uk 10, 24 -- Content-Transfer-Encoding: 8bit 10, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2HDfHuG025454 ==============================End of - Headers==============================
Not only did FEI not "produce the world's first" TEM in 1949 -- they did not manufacture the first commercial one either.
My understanding is that the first commercial TEM was a Metropolitan- Vickers instrument manufactured in 1936 in England. It was shortly followed by a superior microscope from Siemens and Halske in 1939 in Germany. Hillier, Zworykin, and Snyder at RCA were working on electron microscopy in the 1930s and 1940s, putting out their first commercial model in 1940. In the 1940s, Hillier was also outfitting TEMs with EELS and designing the electron microprobe a few years before Castaing.
However, at the FEI website under "A Brief History of FEI", I found the statement: "1949: Philips Electron Optics introduces the world's first commercial transmission electron microscope (TEM)." I, too, am at a loss to explain their claim.
Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
On Mar 17, 2006, at 6:30 AM, richard.beanland-at-bookham.com wrote: } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Browsing through this month's European Semiconductor this morning I } find } an article on page 46 beginning.. } } "Hillsboro (Oregon, USA) based FEI has been in the microscopy business } since 1949, when it produced the world's first Transmission Electron } Microscope (TEM). Today FEI manufactures a full range of } microscopes..." } } Now I'm no history buff, but I didn't see an FEI label on the } replica of } the first TEM in the Science museum in London when I visited last } year.. } How can they make this claim? } } Richard } } ________________________________________ } Richard Beanland } Analytical Services } Bookham Inc } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } } ====================================================================== } = } This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly } prohibited. } No part of this message can be considered a request for goods or } services. } ====================================================================== } = } } } ==============================Original } Headers============================== } 8, 31 -- From richard.beanland-at-bookham.com Fri Mar 17 05:48:23 2006 } 8, 31 -- Received: from mail78.messagelabs.com } (mail78.messagelabs.com [195.245.230.131]) } 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id } k2HBmM8R011251 } 8, 31 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 } 05:48:22 -0600 } 8, 31 -- X-VirusChecked: Checked } 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com } 8, 31 -- X-Msg-Ref: server-5.tower-78.messagelabs.com!1142596059! } 44508703!15 } 8, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- } 8, 31 -- X-Originating-IP: [213.249.209.179] } 8, 31 -- Received: (qmail 13263 invoked from network); 17 Mar 2006 } 11:48:19 -0000 } 8, 31 -- Received: from unknown (HELO cas-smx- } brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 8, 31 -- by server-5.tower-78.messagelabs.com with SMTP; 17 Mar } 2006 11:48:19 -0000 } 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI } ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with } Microsoft SMTPSVC(6.0.3790.211); } 8, 31 -- Fri, 17 Mar 2006 11:48:15 +0000 } 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 8, 31 -- Content-class: urn:content-classes:message } 8, 31 -- MIME-Version: 1.0 } 8, 31 -- Content-Type: text/plain; } 8, 31 -- charset="us-ascii" } 8, 31 -- Subject: The first TEM } 8, 31 -- Date: Fri, 17 Mar 2006 11:48:14 -0000 } 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047C36-at-cas- } smx-02.BOOKHAM.ENTERPRISE.PRI} } 8, 31 -- X-MS-Has-Attach: } 8, 31 -- X-MS-TNEF-Correlator: } 8, 31 -- Thread-Topic: The first TEM } 8, 31 -- Thread-Index: AcZJuKzBYGI+b8m5RBapPz+el04xIw== } 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 8, 31 -- To: {microscopy-at-microscopy.com} } 8, 31 -- X-OriginalArrivalTime: 17 Mar 2006 11:48:15.0713 (UTC) } FILETIME=[AD4FB510:01C649B8] } 8, 31 -- Content-Transfer-Encoding: 8bit } 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k2HBmM8R011251 } ==============================End of - } Headers============================== }
==============================Original Headers============================== 9, 18 -- From frah0010-at-umn.edu Fri Mar 17 08:09:29 2006 9, 18 -- Received: from mtaout-a.tc.umn.edu (mtaout-a.tc.umn.edu [134.84.119.206]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HE9ScH001691 9, 18 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 08:09:28 -0600 9, 18 -- Received: from [160.94.146.133] (212-swing.geo.umn.edu [160.94.146.133]) by mtaout-a.tc.umn.edu with ESMTP; Fri, 17 Mar 2006 08:09:27 -0600 (CST) 9, 18 -- X-Umn-Remote-Mta: [N] 212-swing.geo.umn.edu [160.94.146.133] #+LO+TS+AU+HN 9, 18 -- In-Reply-To: {200603171230.k2HCUvdj018545-at-ns.microscopy.com} 9, 18 -- References: {200603171230.k2HCUvdj018545-at-ns.microscopy.com} 9, 18 -- Mime-Version: 1.0 (Apple Message framework v746.3) 9, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 18 -- Message-Id: {1F3BDA84-09EF-4A54-AEA1-2ACD89920E5D-at-umn.edu} 9, 18 -- Cc: Ellery Frahm {frah0010-at-umn.edu} 9, 18 -- Content-Transfer-Encoding: 7bit 9, 18 -- From: Ellery Frahm {frah0010-at-umn.edu} 9, 18 -- Subject: Re: [Microscopy] The first TEM 9, 18 -- Date: Fri, 17 Mar 2006 08:09:25 -0600 9, 18 -- To: Microscopy-at-microscopy.com 9, 18 -- X-Mailer: Apple Mail (2.746.3) ==============================End of - Headers==============================
You just know there's going to be a few responses to this. So mine is brief.
Yes I've got to agree. I'm sure Ernst Ruska got a Nobel Prize for it (admittedly it took about 50 years) - maybe FEI know something different. First effective design and implementation of a resolution better than a light microscope between 1931 and 1933 then with Siemens first commercial instrument 1939 - again no mention of FEI or Phillips.
I'm sure that by 1949 most industrial countries had the beginnings of commercial instruments.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: richard.beanland-at-bookham.com
{ SEQ CHAPTER \h \r 1}Hello everybody, We want to upgrade of our digital system. We have a digital camera on our H7500. It is a Hamamatsu Advantage HR side mounted one megapixel camera. Is there an advantage of having 6 megapixel versus 4 megapixel camera for biological materials? Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Fri Mar 17 08:49:22 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HEnJ0J015311 1, 22 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 08:49:20 -0600 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1FKGGT-000613-01 1, 22 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 10:49:17 -0400 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 17 Mar 06 10:49:16 -0400 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 17 Mar 06 10:48:50 -0400 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Fri, 17 Mar 2006 10:43:53 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: TEM digital camera 1, 22 -- Message-ID: {441A92E9.14981.3CD146-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
As two of my main interests are photography and microscopes, I have had a more thorough look at the on-line archives to find out who made the first photomicrograph (it was surprisingly easy - articles on photography appear as well represented on the internet as those on the computer). If you are interested please read further :-
X-from 'offline' discussions, it seems that Fox Talbot (or possibly others such as the Rev. J.B. Meade or Daguerre) could claim to have produced the first documented 'fixed' photomicrograph (a magnified image on light sensitive paper). Fox Talbot certainly used a 'solar microscope' for his 'photogenic drawings' in the previous year to Dancer's Flea (shown in 1840). I did find a reference to the 'solar microscope' without having to visit any Museums. It's clearly essentially a camera obscura (lucida) with enlargement 'microscope' lenses to produce magnified images for tracing onto paper or exposing light sensitive paper (e.g. a photograph of 'lace' attributed to Fox Talbot in Sept 1839). I think even that counts as a photomicrograph. Problems with long-term fixing of the image means its not very impressive now. Solar microscopes had been around for a long time as an artists aid, long before Fox Talbot used it to focus images onto his light sensitive paper. They were easily available to those with the money to afford one. I have seen a well made 'solar microscope' example dating from 1750, see http://www.mhs.ox.ac.uk/cameras/index.htm?item18 - this museum in Oxford also an original 1839 Fox Talbot 'lace' photograph (image on-line).
In 1835 Talbot produced the first 'negative' image - the Oriel window in the South Gallery at Lacock Abbey, Wiltshire (near where I live - and it's where he died). This is now the oldest surviving photo (negative) using todays photographic process, although others such as Daguerre were doing similar work at the time (but his method had no means of reproducing the image via the 'negatives'). I can't quickly find any on-line images of Fox Talbot or others more scientific solar microscope 'photogenic drawings', but they are mentioned in the letter from the Rev. J. B. Reade to Fox Talbot suggesting that his photographic use of gallic acid preceded that of Talbot :
"So early as April 1839 the Rev. J.B. Reade made a sensitive paper by using infusion of galls after nitrate of silver; by this process Mr. Reade obtained several drawings of microscopic objects by means of the solar microscope; the drawings were taken before the paper was dry.".
X-from other letters it seems that Talbot had used a solar microscope for photograph enlargements (photomicrographs) before January 1839 (see below). So it's almost certain Talbot (or another early photographer) will have beaten Dancer to producing the first photomicrograph, although as Dancer's 'flea' was 'exhibited' in 1840 he may have made earlier versions. It's also likely that Daguerre and others will have produced magnified images during their experiments from 1935 to 1939, although even Daguerre had problems with fixing the image until 1837.
When Daguerre make public his photographic process on the 7th Jan 1839, Fox Talbot wrote to William Jerdan (Royal Society) on the 30th January 1839 emphasising that his early work was independent of Daguerre and was in no way influenced by Daguerre's research. Fox Talbot wrote: "The Specimens of this art which I exhibited at the Royal Institution, though consisting only of what I happened to have with me in Town, are yet sufficient to give a general idea of it, and to shew the wide range of its applicability. Among them were pictures of flowers and leaves; a pattern of lace; figures taken from painted glass; a view of Venice copied from an engraving, some images formed by the Solar Microscope, viz. a slice of wood very highly magnified, exhibiting the pores of two kinds, one set much smaller than the other, and more numerous. Another Microscopic sketch, exhibiting the reticulations on the wing of an insect. Finally: various pictures, representing the architecture of my house in the country; all these made with the Camera Obscura in the summer of 1835". This conflicts with some reports that Talbot probably purchased his 'solar microscope' in March 1839 (a receipt in his collection isn't specific on what items were bought then). A lot of Fox Talbot correspondence is now online at http://www.foxtalbot.arts.gla.ac.uk.
However, one of the first photographs that Daguerre displayed publicly was of a dead spider seen in the solar microscope. This was cited in a 6th January 1839 letter from H. Gaucheraud that had originally appeared in La Gazette de France; Fox Talbot likely became aware of this when the letter was reprinted in The Literary Gazette, on the 12 January 1839. So the first photomicrograph is about as old as the first photograph. I expect they were all rushing about then looking for novel images to make into photographs - or 'photogenic drawings' as they called them, e.g. Talbot mentions in a letter (2nd Nov 1839) "I saw Cooper at the Polytechnic taking a drawing with the Oxy hydrogen Microscope* in three minutes". *A microscope employing the light produced by the burning of lime under a current of oxyhydrogen gas.
Also Niépce, Daguerre's early collaborator, is universally credited with producing the first successful photograph (heliograph) in June/July 1827. Besides even the image on the medieval Turin shroud appears to be similar to a 'photographic' one - although it may not have been created intentionally. Also many were experimenting before the 1827 date. Indeed Sir Humphrey Davy (1778-1829) the chemist, together with Thomas Wedgwood (1771-1805), son of Josiah Wedgwood the potter, undertook photographic experiments. In 1800 Davy and Wedgwood succeeded in taking a photograph, they did not, however succeed in fixing the image. Indeed years later, Fox Talbot remembered that "the first person who applied photography to the solar microscope was undoubtedly Mr Wedgwood - but none of his delineations have been preserved, and I believe that no particulars are known. Next in order of time to Mr Wedgwood's, came my own experiments. Having published my first photographic process in January, 1839, I immediately applied it to the solar microscope, and in the course of that year made a great many microscopic photographs, which I gave away to Sir John Herschell, Sir Walter Calverley Trevelyan, and other friends". In Sept 8th 1839 Talbot wrote "I have great hopes of this branch of the Art proving very useful, as for instance in copying the forms of minute crystallization which are so complicated as almost to defy the pencil" - he clearly also had an interest in taking photographs of crystals.
So there are many investigators that could have got there first, although most likely their methods of image capture didn't have the 'negative' reproduction capabilities of Talbot's technique and/or their photo-image just faded soon after capture.
If you are interested in more microscope 'victoriana' also have a look at http://www.diatoms.co.uk where Klaus Kemp has turned diatoms (and butterfly scales) back into an art form.
Regards
Keith --------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL Tel: 020 7608 4050 Fax: 020 7608 4034 email: keith.morris-at-ucl.ac.uk
==============================Original Headers============================== 13, 27 -- From keith.morris-at-ucl.ac.uk Fri Mar 17 08:55:20 2006 13, 27 -- Received: from vscani-a.ucl.ac.uk (vscani-a.ucl.ac.uk [144.82.108.29]) 13, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HEtJjr017624 13, 27 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 08:55:20 -0600 13, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 13, 27 -- by vscani-a.ucl.ac.uk with smtp (Exim 4.51) 13, 27 -- id 1FKGMH-0000i5-TH 13, 27 -- for Microscopy-at-microscopy.com; Fri, 17 Mar 2006 14:55:18 +0000 13, 27 -- Message-ID: {005d01c649d2$b2460190$7b865290-at-keithhigrade} 13, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 13, 27 -- To: {Microscopy-at-microscopy.com} 13, 27 -- Subject: Re: [Microscopy] Fw: First photomicrograph 13, 27 -- Date: Fri, 17 Mar 2006 14:54:30 -0000 13, 27 -- MIME-Version: 1.0 13, 27 -- Content-Type: text/plain; 13, 27 -- format=flowed; 13, 27 -- charset="iso-8859-1"; 13, 27 -- reply-type=original 13, 27 -- Content-Transfer-Encoding: 8bit 13, 27 -- X-Priority: 3 13, 27 -- X-MSMail-Priority: Normal 13, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2670 13, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2670 13, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 13, 27 -- X-UCL-MailScanner: Found to be clean 13, 27 -- X-UCL-MailScanner-SpamCheck: 13, 27 -- X-MailScanner-From: keith.morris-at-ucl.ac.uk ==============================End of - Headers==============================
Maybe this is pie in the sky, but perhaps, just perhaps, someone from FEI might address this issue, if they have the courage.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: malcolm.haswell-at-sunderland.ac.uk [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Friday, March 17, 2006 10:37 AM To: kenconverse-at-qualityimages.biz
Richard
You just know there's going to be a few responses to this. So mine is brief.
Yes I've got to agree. I'm sure Ernst Ruska got a Nobel Prize for it (admittedly it took about 50 years) - maybe FEI know something different. First effective design and implementation of a resolution better than a light microscope between 1931 and 1933 then with Siemens first commercial instrument 1939 - again no mention of FEI or Phillips.
I'm sure that by 1949 most industrial countries had the beginnings of commercial instruments.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: richard.beanland-at-bookham.com
Hi, Ralph
The new NTegra Tomo provides an interesting solution. It combines AFM directly into a Leica microtome so that you can do sections of an appropriate depth, but use the AFM for imaging. We have used this sort of system very successfully already for imaging, sizing and counting hard particles in polymer matrices (silica in PS/HIPS blend). The differences in the local elasticity will image the lung tissue without having to enhance the nanoparticles and, clearly, the difference in hardness will image the nanoparticles. We can have samples run if you would like.
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At 12:26 PM 3/15/2006, rcommon-at-msu.edu wrote:
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==============================Original Headers============================== 16, 17 -- From bfoster-at-mme1.com Fri Mar 17 12:36:04 2006 16, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 16, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HIa31g003592 16, 17 -- for {microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 12:36:03 -0600 16, 17 -- Received: (qmail 25814 invoked from network); 17 Mar 2006 13:37:37 -0500 16, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 16, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 17 Mar 2006 13:37:37 -0500 16, 17 -- Message-Id: {7.0.1.0.0.20060317123205.01ffece0-at-mme1.com} 16, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 16, 17 -- Date: Fri, 17 Mar 2006 12:36:08 -0600 16, 17 -- To: rcommon-at-msu.edu, Microscopy Listserver {microscopy-at-microscopy.com} 16, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 16, 17 -- Subject: Re: [Microscopy] Nanogold in semithin sections 16, 17 -- In-Reply-To: {200603151826.k2FIQXtO001828-at-ns.microscopy.com} 16, 17 -- References: {200603151826.k2FIQXtO001828-at-ns.microscopy.com} 16, 17 -- Mime-Version: 1.0 16, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
The April, 2003, issue of Microscopy and Microanalysis contains an article, "Key Events in the History of Electron Microscopy" which states that in 1937, "The Metropolitan Vickers Company (Manchester, UK) supplies the first commercial electron microscope to Louis C. Martin at Imperial College, London". By way of trivia - The B.F. Goodrich Company (Akron, Ohio) purchased a Philips EM100B in 1957, the delivery of which was conditional upon the approval of W. Ladd whom BFG hired to check out the instrument which was assembled for just that purpose at the Philips' facilities in Mt. Vernon, NY. My...how times have changed!
Ron Smith, Lake Havasu City, AZ
==============================Original Headers============================== 3, 17 -- From Ron2450-at-aol.com Fri Mar 17 12:36:42 2006 3, 17 -- Received: from imo-m28.mx.aol.com (imo-m28.mx.aol.com [64.12.137.9]) 3, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HIae6V003713 3, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 12:36:41 -0600 3, 17 -- Received: from Ron2450-at-aol.com 3, 17 -- by imo-m28.mx.aol.com (mail_out_v38_r7.3.) id w.2a4.76b7015 (17079) 3, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Mar 2006 13:36:34 -0500 (EST) 3, 17 -- From: Ron2450-at-aol.com 3, 17 -- Message-ID: {2a4.76b7015.314c5bb1-at-aol.com} 3, 17 -- Date: Fri, 17 Mar 2006 13:36:33 EST 3, 17 -- Subject: Re: First Commercial TEM 3, 17 -- To: Microscopy-at-microscopy.com 3, 17 -- MIME-Version: 1.0 3, 17 -- Content-Type: text/plain; charset="US-ASCII" 3, 17 -- Content-Transfer-Encoding: 7bit 3, 17 -- X-Mailer: 9.0 SE for Windows sub 5022 3, 17 -- X-Spam-Flag: NO ==============================End of - Headers==============================
Hi group - this is what I found when I did a Google search: Early experiments using X-rays of extremely short wavelength were not pursued further because of the inability to focus these rays. The first breakthrough in the development of the electron microscope came when Louis de Broglie advanced his theory that the electron had a dual nature, with characteristics of a particle or a wave. The demonstration, in 1923 by Busch, that a beam of electrons could be focused by magnetic or electric fields opened the way for the development of the first electron microscope, in 1932, by Knoll and Ruska. Although the initial development of the electron microscope, in Germany, was followed by technical improvements in America, the first commercially available apparatus was marketed by Seimens.
==============================Original Headers============================== 1, 18 -- From maloneyb-at-fiu.edu Fri Mar 17 15:40:58 2006 1, 18 -- Received: from smtp10.fiu.edu (smtp10.fiu.edu [131.94.79.27]) 1, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HLevp5024909 1, 18 -- for {microscopy-at-ns.microscopy.com} ; Fri, 17 Mar 2006 15:40:57 -0600 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu [131.94.95.156]) 1, 18 -- by smtp10.fiu.edu (MOS 3.7.1-GA) 1, 18 -- with ESMTP id CLW05883; 1, 18 -- Fri, 17 Mar 2006 16:40:56 -0500 (EST) 1, 18 -- Message-ID: {441B2B93.6060308-at-fiu.edu} 1, 18 -- Date: Fri, 17 Mar 2006 16:35:15 -0500 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 18 -- X-Accept-Language: en-us, en 1, 18 -- MIME-Version: 1.0 1, 18 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-ns.microscopy.com} 1, 18 -- Subject: first TEM 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Contrary to common impression, it appears Ruska and Knoll were not even aware of de Broglie's work when they started trying to build an electron microscope, in fact they were initially not happy to hear about it because they had hoped to escape the limits imposed by wave optics! In his Nobel lecture in 1988 Ruska said:
"As engineers we did not know yet the thesis on the "material waves" by French physicist de Broglie that had been put forward several years earlier (1925). . . When I first heard of it in summer 1931, I was very much disappointed that the resolution should now be limited again by a wavelength (of the materiestrahlung). I was immediately heartened, though, when with the aid of the de Broglie equation I became satisfied that these waves must be around five orders of magnitude shorter in length than light waves. Thus, there was no reason to abandon the aim of electron microscopy surpassing the resolution of light microscopy".
Other interesting facts can be found in "EMSA and Its people the First Fifty Years" by Sterling Newberry and published by the Electron Microscopy Society of America in 1992.
Marie
On Mar 17, 2006, at 5:12 PM, maloneyb-at-fiu.edu wrote:
} Early experiments using X-rays of extremely short wavelength were not } pursued further because of the inability to focus these rays. The first } breakthrough in the development of the electron microscope came when } Louis de Broglie advanced his theory that the electron had a dual } nature, with characteristics of a particle or a wave. The } demonstration, } in 1923 by Busch, that a beam of electrons could be focused by magnetic } or electric fields opened the way for the development of the first } electron microscope, in 1932, by Knoll and Ruska. Although the initial } development of the electron microscope, in Germany, was followed by } technical improvements in America, the first commercially available } apparatus was marketed by Seimens. } } ==============================Original } Headers============================== } 1, 18 -- From maloneyb-at-fiu.edu Fri Mar 17 15:40:58 2006 } 1, 18 -- Received: from smtp10.fiu.edu (smtp10.fiu.edu [131.94.79.27]) } 1, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } k2HLevp5024909 } 1, 18 -- for {microscopy-at-ns.microscopy.com} ; Fri, 17 Mar 2006 } 15:40:57 -0600 } 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu } [131.94.95.156]) } 1, 18 -- by smtp10.fiu.edu (MOS 3.7.1-GA) } 1, 18 -- with ESMTP id CLW05883; } 1, 18 -- Fri, 17 Mar 2006 16:40:56 -0500 (EST) } 1, 18 -- Message-ID: {441B2B93.6060308-at-fiu.edu} } 1, 18 -- Date: Fri, 17 Mar 2006 16:35:15 -0500 } 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} } 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; } rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) } 1, 18 -- X-Accept-Language: en-us, en } 1, 18 -- MIME-Version: 1.0 } 1, 18 -- To: "'microscopy-at-msa.microscopy.com'" } {microscopy-at-ns.microscopy.com} } 1, 18 -- Subject: first TEM } 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed } 1, 18 -- Content-Transfer-Encoding: 7bit } ==============================End of - } Headers============================== } } Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 8, 21 -- From marie.cantino-at-uconn.edu Sat Mar 18 09:07:48 2006 8, 21 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 8, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2IF7lbP015649 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 09:07:48 -0600 8, 21 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 8, 21 -- by mail1.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2IF7eK11604 8, 21 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 10:07:40 -0500 8, 21 -- Mime-Version: 1.0 (Apple Message framework v623) 8, 21 -- In-Reply-To: {200603172212.k2HMClqn030823-at-ns.microscopy.com} 8, 21 -- References: {200603172212.k2HMClqn030823-at-ns.microscopy.com} 8, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 8, 21 -- Message-Id: {e5a8d5e1aab9c76980512131f1fbaff6-at-uconn.edu} 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 8, 21 -- Subject: Re: [Microscopy] first TEM 8, 21 -- Date: Sat, 18 Mar 2006 10:07:40 -0500 8, 21 -- To: Microscopy-at-Microscopy.Com 8, 21 -- X-Mailer: Apple Mail (2.623) 8, 21 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 8, 21 -- X-UConn-MailScanner: Found to be clean 8, 21 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
Has anyone else had problems in negative staining using recent bottles of uranyl acetate? None of our newer bottles (we have purchased several from several different vendors in the past year) work as well as one purchased in 1991. Problems include poor spreading and precipitation or aggregation of the stain. The newer stuff is much more soluble than the old, has a strong odor of acetic acid, and is lighter in color. Some of the new bottles work better than others (though none as well as the old stuff), but there was no correlation with any variable except pH. Lots that worked best had higher pH in solution (3.5-3.8) than poor lots (2.3-2.5), but raising the pH of the solution did not produce better staining. Results were quite variable between lots from the same vendor. As far as I can tell, all lots work OK for positive staining of resin sections (though we don't want to waste any of the "best" bottle making tests on sections!)
We're wondering whether anyone else has noticed this and found a better source or a way to improve the existing lots.
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 5, 19 -- From marie.cantino-at-uconn.edu Sat Mar 18 09:37:07 2006 5, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2IFb6KM021362 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 09:37:06 -0600 5, 19 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 5, 19 -- by mail2.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2IFb1l15528 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Sat, 18 Mar 2006 10:37:01 -0500 5, 19 -- Mime-Version: 1.0 (Apple Message framework v623) 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- Message-Id: {d84f87dd1f62e47529f153e4ea711649-at-uconn.edu} 5, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 5, 19 -- To: Microscopy-at-Microscopy.Com 5, 19 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 5, 19 -- Subject: Uranyl acetate/negative staining problems 5, 19 -- Date: Sat, 18 Mar 2006 10:37:00 -0500 5, 19 -- X-Mailer: Apple Mail (2.623) 5, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 5, 19 -- X-UConn-MailScanner: Found to be clean 5, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
I'm not sure if you got your question answered yet.
Here are a few questions for you to help fill in some gaps.
What feature size(s) are you dealing with? Why are you using a Ta substrate?
I suspect that any TaN etchant will also etch the Ta. Thus, there is no stop mechanism for etching just the TaN.
TaN is sometimes put on top of Cu damascene runners. These are plasma etched using an oxidizing plasma. If you want to use wet etch, I would change the substrate to something that is not going to be etched, or coat whatever you have with Si3N4 which is a stop. Then, use Transene Etch III.
If your feature sizes are too small, wet etch won't do a good job, IMO. If the resist is thick enough and cured, any type should work. Which type you use is going to depend on UV wavelength and what is used to pattern/expose the resist as well as compatable developer.
gary g.
At 11:34 AM 3/15/2006, you wrote:
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==============================Original Headers============================== 13, 21 -- From gary-at-gaugler.com Sat Mar 18 12:04:42 2006 13, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2II4eGY004349 13, 21 -- for {microscopy-at-microscopy.com} ; Sat, 18 Mar 2006 12:04:41 -0600 13, 21 -- Received: (qmail 4270 invoked from network); 18 Mar 2006 10:04:38 -0800 13, 21 -- Received: by simscan 1.1.0 ppid: 4267, pid: 4268, t: 0.1675s 13, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 21 -- by qsmtp3 with SMTP; 18 Mar 2006 10:04:38 -0800 13, 21 -- Message-Id: {6.2.3.4.2.20060318095716.0205d038-at-mail.calweb.com} 13, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 13, 21 -- Date: Sat, 18 Mar 2006 10:04:39 -0800 13, 21 -- To: ramadanhany-at-gmail.com 13, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 21 -- Subject: Re: [Microscopy] Tantlum oxide etching aginst photoresist 13, 21 -- material 13, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 21 -- In-Reply-To: {200603151934.k2FJYCTe014754-at-ns.microscopy.com} 13, 21 -- References: {200603151934.k2FJYCTe014754-at-ns.microscopy.com} 13, 21 -- Mime-Version: 1.0 13, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Um... might be safer to say North America than just America, if it is Hillier's work you're thinking of. The group he was in made a practical TEM at the University of Toronto in 1938 (if I recall correctly), and he didn't move to the US until 1940.
Mike Fay School of Mechanical, Materials and Manufacturing Engineering Nottingham University University Park Nottingham NG7 2RD tel 0115 8466081
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Hi group - this is what I found when I did a Google search: Early experiments using X-rays of extremely short wavelength were not pursued further because of the inability to focus these rays. The first breakthrough in the development of the electron microscope came when Louis de Broglie advanced his theory that the electron had a dual nature, with characteristics of a particle or a wave. The demonstration, in 1923 by Busch, that a beam of electrons could be focused by magnetic or electric fields opened the way for the development of the first electron microscope, in 1932, by Knoll and Ruska. Although the initial development of the electron microscope, in Germany, was followed by technical improvements in America, the first commercially available apparatus was marketed by Seimens.
==============================Original Headers============================== 1, 18 -- From maloneyb-at-fiu.edu Fri Mar 17 15:40:58 2006 1, 18 -- Received: from smtp10.fiu.edu (smtp10.fiu.edu [131.94.79.27]) 1, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2HLevp5024909 1, 18 -- for {microscopy-at-ns.microscopy.com} ; Fri, 17 Mar 2006 15:40:57 -0600 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu [131.94.95.156]) 1, 18 -- by smtp10.fiu.edu (MOS 3.7.1-GA) 1, 18 -- with ESMTP id CLW05883; 1, 18 -- Fri, 17 Mar 2006 16:40:56 -0500 (EST) 1, 18 -- Message-ID: {441B2B93.6060308-at-fiu.edu} 1, 18 -- Date: Fri, 17 Mar 2006 16:35:15 -0500 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 18 -- X-Accept-Language: en-us, en 1, 18 -- MIME-Version: 1.0 1, 18 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-ns.microscopy.com} 1, 18 -- Subject: first TEM 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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==============================Original Headers============================== 13, 24 -- From michael.fay-at-nottingham.ac.uk Mon Mar 20 04:57:24 2006 13, 24 -- Received: from haydn.is.nottingham.ac.uk (haydn.is.nottingham.ac.uk [128.243.40.92]) 13, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KAvN7J001360 13, 24 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 04:57:23 -0600 13, 24 -- Received: from ccw0m1.ccc.nottingham.ac.uk ([128.243.220.65] helo=ccw0m1.nottingham.ac.uk) 13, 24 -- by haydn.is.nottingham.ac.uk with esmtp (Exim 3.36 #2) 13, 24 -- id 1FLI4X-0002DA-00 13, 24 -- for microscopy-at-microscopy.com; Mon, 20 Mar 2006 10:57:13 +0000 13, 24 -- Received: from Gwweb1-MTA by ccw0m1.nottingham.ac.uk 13, 24 -- with Novell_GroupWise; Mon, 20 Mar 2006 10:57:14 +0000 13, 24 -- Message-Id: {s41e8a8a.006-at-ccw0m1.nottingham.ac.uk} 13, 24 -- X-Mailer: Novell GroupWise Internet Agent 6.0.3 13, 24 -- Date: Mon, 20 Mar 2006 10:57:01 +0000 13, 24 -- From: "Michael Fay" {Michael.Fay-at-nottingham.ac.uk} 13, 24 -- To: {microscopy-at-microscopy.com} 13, 24 -- Subject: Re: [Microscopy] first TEM 13, 24 -- Mime-Version: 1.0 13, 24 -- Content-Type: text/plain; charset=US-ASCII 13, 24 -- Content-Disposition: inline 13, 24 -- X-MailScanner-Information: Please contact staff-it-helpline-at-nottingham.ac.uk for more information 13, 24 -- X-MailScanner: Found to be clean 13, 24 -- X-MailScanner-From: michael.fay-at-nottingham.ac.uk 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2KAvN7J001360 ==============================End of - Headers==============================
Sorry to bother the list with this, but one person responding to my question about UA left a phone message for me, which I accidentally deleted before writing down the name and correct phone number. If you are that person, could you please call or e-mail again? Thanks!
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 4, 19 -- From marie.cantino-at-uconn.edu Mon Mar 20 10:20:11 2006 4, 19 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KGKAjZ016191 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 10:20:11 -0600 4, 19 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 4, 19 -- by mail1.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2KGA0K14987 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- Message-Id: {f8d53214ee4a81c62c984e77fbf3b665-at-uconn.edu} 4, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 19 -- To: microscopy-at-Microscopy.Com 4, 19 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 4, 19 -- Subject: Uranyl acetate/negative staining problems 4, 19 -- Date: Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- X-Mailer: Apple Mail (2.623) 4, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 4, 19 -- X-UConn-MailScanner: Found to be clean 4, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (danielluth-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 16, 2006 at 19:01:41 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both danielluth-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: danielluth-at-yahoo.com Name: Daniel Luth
Organization: University of Melbourne
Education: Graduate College
Location: Melbourne, Victoria, Australia
Title: Are parasites detectable in the blood from a microscope?
Question: Hi, My wife recently went to a naturopath who had a degree in Microscopy, and took a sample of her blood to view under the microscope. The image was projected onto a monitor. The naturopath was able to detect parasites in the blood - and pointed them out. My wife described them as wiggly moving threads. My question is are parasites detectable in the blood from a microscope?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ml687644-at-bigpond.net.au) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 17, 2006 at 19:28:50 ---------------------------------------------------------------------------
Email: ml687644-at-bigpond.net.au Name: Mark LESTER
Organization: New South Wales Police Service
Education: Undergraduate College
Location: Sydney, Australia
Question: Hi there. I am trying to find out the technique you would use for optimum visualisation and identification of the following specimens;
a)bullet for rifling marks b)sperm cells c)hair d)natural and synthetic fibres e)flies f)spiracles on maggots g)paint flakes (for counting pain layers to trace paint source) h)cannabis leaf to identify cystolithic hairs i)vomit for identifying food particles j)a soil sample k)gunshot residue
if you could outline the optical system used and the appropriate magnification, that would be of great assistance to me.
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Title-Subject: [Filtered] Replica Materials other than Cellulose Acetate for SEM Analysis of Metal Surfaces
Question: What recommendations would you have for replicating materials, other than cellulose acetate, for making replicas of flat metal surfaces that would be used for subsequent viewing and EDS analysis with SEM?
I am interested in any techniques that you might be using, literature citations and/or vendors of replicating materials other than cellulose acetate for materials and biological applications.
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Email: amich-at-ufl.edu Name: Albina
Organization: UF
Title-Subject: [Filtered] vapor fixation
Question: Hi, I would appreciate your suggestions on the following: I work on the project studying bacterial attachment/interaction with various substrates. Some of those substrates are polymers and fibers which cannot withstand standard fixation by immersion since in some cases I would like to avoid wetting the fibers (they swell). I wanted to try vapor fixation; but since it pilot project I am reluctant to pump osmium vapor with pump. Can anyone suggest a way to vapor-fix samples with homemade ěon-the-cheapî setup? It is also my understanding that I would need to vapor fix with osmium first and follow with glutaraldehyde, right? Thank you for consideration, Albina
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Email: b.ambrose-at-massey.ac.nz Name: Barbara Ambrose
Organization: Massey University
Title-Subject: [Filtered] The Ideal Microscopy Center
Question: Dear All We have just secured a large grant to fund a microscopy suite (MMIC) here in New Zealand. MMIC would house a spinning disc confocal microscope, eSEM, TEM, light microscopy and all the processing equipment. We are having to renovate a new space for the microscopy suite and am wondering if you have any advice/suggestions or if you could redesign your suite what you would incorporate or change. Thank you for your time, cheers barbara
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Email: pekysar-at-ucdavis.edu Name: Pat Kysar
Organization: University of California, Davis
Title-Subject: [Filtered] Thanks for Low Dose Info
Question: Thank you to all who responded to my questions about low dose TEM! I have some very good ideas/suggestions. We are very lucky to have such a great resouce at our disposal! Pat Kysar University of California, Davis Medical School, Pathology EM Lab
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Email: Paul.Leahy-at-epa.vic.gov.au Name: Paul Leahy
Organization: EPA Victoria
Title-Subject: [Filtered] Field Microscopes
Question: I'm after a field microscope for looking at small algae. I've come accross the Swift FM-31 and one from Richardson Technologies. Can anyone comment on these? Also does anyone know of suppliers in Australia?
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Email: Paul.Leahy-at-epa.vic.gov.au Name: Paul Leahy
Organization: EPA Victoria
Title-Subject: [Filtered] Field Microscopes
Question: I'm after a field microscope for looking at small algae. I've come accross the Swift FM-31 and one from Richardson Technologies. Can anyone comment on these? Also does anyone know of suppliers in Australia?
Sorry to bother the list with this, but one person responding to my question about UA left a phone message for me, which I accidentally deleted before writing down the name and correct phone number. If you are that person, could you please call or e-mail again? Thanks!
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 4, 19 -- From marie.cantino-at-uconn.edu Mon Mar 20 11:29:22 2006 4, 19 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 4, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KGKAjZ016191 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 10:20:11 -0600 4, 19 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 4, 19 -- by mail1.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id k2KGA0K14987 4, 19 -- for {microscopy-at-Microscopy.Com} ; Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- Message-Id: {f8d53214ee4a81c62c984e77fbf3b665-at-uconn.edu} 4, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 19 -- To: microscopy-at-microscopy.com 4, 19 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 4, 19 -- Subject: Uranyl acetate/negative staining problems 4, 19 -- Date: Mon, 20 Mar 2006 11:10:00 -0500 4, 19 -- X-Mailer: Apple Mail (2.623) 4, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 4, 19 -- X-UConn-MailScanner: Found to be clean 4, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
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Email: pekysar-at-ucdavis.edu Name: Pat Kysar
Organization: University of California, Davis
Title-Subject: [Filtered] Thanks for Low Dose Info
Question: Thank you to all who responded to my questions about low dose TEM! I have some very good ideas/suggestions. We are very lucky to have such a great resouce at our disposal! Pat Kysar University of California, Davis Medical School, Pathology EM Lab
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Email: b.ambrose-at-massey.ac.nz Name: Barbara Ambrose
Organization: Massey University
Title-Subject: [Filtered] The Ideal Microscopy Center
Question: Dear All We have just secured a large grant to fund a microscopy suite (MMIC) here in New Zealand. MMIC would house a spinning disc confocal microscope, eSEM, TEM, light microscopy and all the processing equipment. We are having to renovate a new space for the microscopy suite and am wondering if you have any advice/suggestions or if you could redesign your suite what you would incorporate or change. Thank you for your time, cheers barbara
Out of fairness, I wanted to point out FEI has changed its website to reflect our discussion here:
On Friday it read:
"1949: Philips Electron Optics introduces the world's first commercial transmission electron microscope (TEM)."
Today it reads:
"1949: Philips Electron Optics (part of FEI Company since 1997) was one of the first companies to start volume production of Transmission Electron Microscopes in 1949."
Thumbs up to FEI for changing their website so quickly when the error was pointed out.
Best, Ellery
-------------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
==============================Original Headers============================== 8, 17 -- From frah0010-at-umn.edu Mon Mar 20 11:40:13 2006 8, 17 -- Received: from mtaout-w.tc.umn.edu (mtaout-w.tc.umn.edu [160.94.160.21]) 8, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KHeDUd009714 8, 17 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 11:40:13 -0600 8, 17 -- Received: from [160.94.146.133] (212-swing.geo.umn.edu [160.94.146.133]) by mtaout-w.tc.umn.edu with ESMTP for Microscopy-at-microscopy.com; Mon, 20 Mar 2006 11:39:20 -0600 (CST) 8, 17 -- X-Umn-Remote-Mta: [N] 212-swing.geo.umn.edu [160.94.146.133] #+LO+TS+AU+HN 8, 17 -- Mime-Version: 1.0 (Apple Message framework v746.3) 8, 17 -- In-Reply-To: {200603171731.k2HHVe4l032728-at-ns.microscopy.com} 8, 17 -- References: {200603171731.k2HHVe4l032728-at-ns.microscopy.com} 8, 17 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 17 -- Message-Id: {6A92218F-31CC-4C03-B890-00A51AB2043D-at-umn.edu} 8, 17 -- Content-Transfer-Encoding: 7bit 8, 17 -- From: Ellery Frahm {frah0010-at-umn.edu} 8, 17 -- Subject: Re: [Microscopy] The first TEM 8, 17 -- Date: Mon, 20 Mar 2006 11:39:17 -0600 8, 17 -- To: Microscopy-at-microscopy.com 8, 17 -- X-Mailer: Apple Mail (2.746.3) ==============================End of - Headers==============================
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Title-Subject: [Filtered] Replica Materials other than Cellulose Acetate for SEM Analysis of Metal Surfaces
Question: What recommendations would you have for replicating materials, other than cellulose acetate, for making replicas of flat metal surfaces that would be used for subsequent viewing and EDS analysis with SEM?
I am interested in any techniques that you might be using, literature citations and/or vendors of replicating materials other than cellulose acetate for materials and biological applications.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ml687644-at-bigpond.net.au) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 17, 2006 at 19:28:50 ---------------------------------------------------------------------------
Email: ml687644-at-bigpond.net.au Name: Mark LESTER
Organization: New South Wales Police Service
Education: Undergraduate College
Location: Sydney, Australia
Question: Hi there. I am trying to find out the technique you would use for optimum visualisation and identification of the following specimens;
a)bullet for rifling marks b)sperm cells c)hair d)natural and synthetic fibres e)flies f)spiracles on maggots g)paint flakes (for counting pain layers to trace paint source) h)cannabis leaf to identify cystolithic hairs i)vomit for identifying food particles j)a soil sample k)gunshot residue
if you could outline the optical system used and the appropriate magnification, that would be of great assistance to me.
This Question was submitted to Ask-A-Microscopist by (danielluth-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 16, 2006 at 19:01:41 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both danielluth-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: danielluth-at-yahoo.com Name: Daniel Luth
Organization: University of Melbourne
Education: Graduate College
Location: Melbourne, Victoria, Australia
Title: Are parasites detectable in the blood from a microscope?
Question: Hi, My wife recently went to a naturopath who had a degree in Microscopy, and took a sample of her blood to view under the microscope. The image was projected onto a monitor. The naturopath was able to detect parasites in the blood - and pointed them out. My wife described them as wiggly moving threads. My question is are parasites detectable in the blood from a microscope?
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Email: amich-at-ufl.edu Name: Albina
Organization: UF
Title-Subject: [Filtered] vapor fixation
Question: Hi, I would appreciate your suggestions on the following: I work on the project studying bacterial attachment/interaction with various substrates. Some of those substrates are polymers and fibers which cannot withstand standard fixation by immersion since in some cases I would like to avoid wetting the fibers (they swell). I wanted to try vapor fixation; but since it pilot project I am reluctant to pump osmium vapor with pump. Can anyone suggest a way to vapor-fix samples with homemade ěon-the-cheapî setup? It is also my understanding that I would need to vapor fix with osmium first and follow with glutaraldehyde, right? Thank you for consideration, Albina
African trypanosomes would be detectable with a light microscope and would look like worms moving between the red blood cells, but I don't think sleeping sickness has reached Australia yet.
You could probably detect Giardia in unstained specimens but they are round cells.
What was the final diagnosis and treatment/
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
Email: danielluth-at-yahoo.com Name: Daniel Luth
Organization: University of Melbourne
Education: Graduate College
Location: Melbourne, Victoria, Australia
Title: Are parasites detectable in the blood from a microscope?
Question: Hi, My wife recently went to a naturopath who had a degree in Microscopy, and took a sample of her blood to view under the microscope. The image was projected onto a monitor. The naturopath was able to detect parasites in the blood - and pointed them out. My wife described them as wiggly moving threads. My question is are parasites detectable in the blood from a microscope?
==============================Original Headers============================== 16, 22 -- From PWebster-at-hei.org Mon Mar 20 11:58:03 2006 16, 22 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 16, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KHw3IG028454 16, 22 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 11:58:03 -0600 16, 22 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 16, 22 -- Mon, 20 Mar 2006 17:58:02 +0000 16, 22 -- User-Agent: Microsoft-Entourage/11.2.1.051004 16, 22 -- Date: Mon, 20 Mar 2006 09:58:00 -0800 16, 22 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: parasites 16, 22 -- detectable in the blood 16, 22 -- From: "Webster, Paul" {PWebster-at-hei.org} 16, 22 -- To: {danielluth-at-yahoo.com} 16, 22 -- CC: {microscopy-at-microscopy.com} 16, 22 -- Message-ID: {C0442D28.8E6E%PWebster-at-hei.org} 16, 22 -- Thread-Topic: [Microscopy] [Filtered] AskAMicroscopist: parasites 16, 22 -- detectable in the blood 16, 22 -- Thread-Index: AcZMR9NqEcoBzrg7EdqcYAANk7Zh7g== 16, 22 -- In-Reply-To: {200603201753.k2KHrSUE017918-at-ns.microscopy.com} 16, 22 -- Mime-version: 1.0 16, 22 -- Content-type: text/plain; 16, 22 -- charset="US-ASCII" 16, 22 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
Yes, there are parasites that show up on blood films, usually with a special stain. Live wiggling things might easily be white blood cells doing their normal amoboid movement or just brownian motion. I for one would not trust my diagnosis to a "naturopath who had a degree in Microscopy". This sounds like quackery to me. There is a school of (what I consider charlatans) practioners that diagnose everything from blood smears. If your wife has a parasitic infection she will have distinct symptoms. Do you have objections to physicians with real medical training?
Geoff
danielluth-at-yahoo.com wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 7, 32 -- From mcauliff-at-umdnj.edu Mon Mar 20 11:58:44 2006 7, 32 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 7, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KHwhIP030774 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 11:58:44 -0600 7, 32 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 7, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 9192A4BE1D 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 12:58:43 -0500 (EST) 7, 32 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 7, 32 -- by zix03.umdnj.edu (Proprietary) with ESMTP id D9EC54BE03 7, 32 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 12:58:42 -0500 (EST) 7, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 7, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 7, 32 -- id {0IWF00L01T3NCO-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 7, 32 -- for microscopy-at-msa.microscopy.com; Mon, 20 Mar 2006 12:58:42 -0500 (EST) 7, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 7, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 7, 32 -- 2004)) with ESMTP id {0IWF00AV7TXAGV-at-Polaris.umdnj.edu} ; Mon, 7, 32 -- 20 Mar 2006 12:58:23 -0500 (EST) 7, 32 -- Date: Mon, 20 Mar 2006 12:57:27 -0500 7, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 7, 32 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: parasites detectable in 7, 32 -- the blood 7, 32 -- In-reply-to: {200603201705.k2KH5IXo029145-at-ns.microscopy.com} 7, 32 -- To: danielluth-at-yahoo.com, MicroscopyListserver {microscopy-at-msa.microscopy.com} 7, 32 -- Message-id: {441EED07.2080201-at-umdnj.edu} 7, 32 -- MIME-version: 1.0 7, 32 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 7, 32 -- Content-transfer-encoding: 7BIT 7, 32 -- X-Accept-Language: en-us, en 7, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 7, 32 -- Gecko/20040804 Netscape/7.2 (ax) 7, 32 -- References: {200603201705.k2KH5IXo029145-at-ns.microscopy.com} ==============================End of - Headers==============================
Julie Duimstra wrote: =========================================================== Title-Subject: [Filtered] Replica Materials other than Cellulose Acetate for SEM Analysis of Metal Surfaces
Question: What recommendations would you have for replicating materials, other than cellulose acetate, for making replicas of flat metal surfaces that would be used for subsequent viewing and EDS analysis with SEM?
I am interested in any techniques that you might be using, literature citations and/or vendors of replicating materials other than cellulose acetate for materials and biological applications =========================================================== Could you tell us what it is that you find objectionable to cellulose acetate? That information would be helpful in determining that might be a menu of possible alternatives. Normally on a metal surface, CA works just fine.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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==============================Original Headers============================== 12, 25 -- From cgarber-at-2spi.com Mon Mar 20 13:09:37 2006 12, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 12, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KJ9buN017167 12, 25 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 13:09:37 -0600 12, 25 -- Received: from ibm1x23g2abfyg ([70.5.4.200]) 12, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k2KJ8wrV032355; 12, 25 -- Mon, 20 Mar 2006 14:09:03 -0500 12, 25 -- X-IDV-FirstRcvd: [70.5.4.200] 12, 25 -- X-IDV-HELO: ibm1x23g2abfyg 12, 25 -- Message-ID: {006501c64c51$bef55950$c8040546-at-ibm1x23g2abfyg} 12, 25 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 12, 25 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 12, 25 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 25 -- Cc: {julie.duimstra-at-hti.hutch.com} 12, 25 -- Subject: Alternatives to cellulose acetate as a replicating resin 12, 25 -- Date: Mon, 20 Mar 2006 14:08:15 -0500 12, 25 -- MIME-Version: 1.0 12, 25 -- Content-Type: text/plain; 12, 25 -- charset="Windows-1252" 12, 25 -- X-Priority: 3 12, 25 -- X-MSMail-Priority: Normal 12, 25 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 12, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 12, 25 -- Content-Transfer-Encoding: 8bit 12, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2KJ9buN017167 ==============================End of - Headers==============================
You don't give us the reason you are looking for an alternative to the acetate replicas, or much detail about what you are trying to do.
You talk about EDS analysis. Are you perhaps trying to extract particles from the surface and analyze them? If so, then a very common technique 20 years ago was the carbon extraction replica. This involves lightly etching a surface to get the particles standing proud, then evaporating a layer of carbon, and finally a deeper etch to release the particles - now embedded in the carbon film, which is floated off on the surface of water. The film is picked up on a grid and examined in the TEM or SEM - depending on the particles. For applications where carbon is not appropriate, other materials can be used (to my knowledge, Al and amorphous SiO have been used, and possibly others).
In a variation (dating to even earlier in the history of microscopy), the replica would have a light "shadow" of a metal like Pt evaporated on it at an oblique angle, to enhance visualization of the topography of the surface. This was often used to allow surface topography to be investigated in the TEM, before SEMs were common (which, I hasten to add, was before I was active in the field!)
If this is anything like what you have in mind, I would be happy to expand on the procedure off-line.
Tony Garratt-Reed.
At 12:46 PM 3/20/2006, you wrote:
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*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
You could use vapors from warm or hot paraformaldehyde (the solid) or acrolein (a liquid) or osmium. Note that osmium can be dissolved in solvents other than water. Or perhaps follow one of the first two with osmium for lipid staining. You don't have to pump osmiun, just put the object of interest in a closed container with the fixative of your choice, the put the container under a fume hood.
Geoff
amich-at-ufl.edu wrote:
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==============================Original Headers============================== 8, 31 -- From mcauliff-at-umdnj.edu Mon Mar 20 15:23:14 2006 8, 31 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 8, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KLNEF2006472 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 15:23:14 -0600 8, 31 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 8, 31 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 1A2AE4BE66 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 15:23:14 -0600 (CST) 8, 31 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 8, 31 -- by zix02.umdnj.edu (Proprietary) with ESMTP id D30CE4BE38 8, 31 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Mar 2006 15:23:12 -0600 (CST) 8, 31 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 31 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 31 -- id {0IWG00H012D4JC-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 31 -- for microscopy-at-msa.microscopy.com; Mon, 20 Mar 2006 16:23:12 -0500 (EST) 8, 31 -- Received: from [127.0.0.1] ([10.138.2.240]) 8, 31 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 31 -- 2004)) with ESMTP id {0IWG00BM23EDKD-at-Polaris.umdnj.edu} ; Mon, 8, 31 -- 20 Mar 2006 16:23:01 -0500 (EST) 8, 31 -- Date: Mon, 20 Mar 2006 16:22:06 -0500 8, 31 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 31 -- Subject: Re: [Microscopy] viaWWW: vapor fixation 8, 31 -- In-reply-to: {200603201709.k2KH9SMU030393-at-ns.microscopy.com} 8, 31 -- To: amich-at-ufl.edu, MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 31 -- Message-id: {441F1CFE.2030102-at-umdnj.edu} 8, 31 -- MIME-version: 1.0 8, 31 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 8, 31 -- Content-transfer-encoding: 8BIT 8, 31 -- X-Accept-Language: en-us, en 8, 31 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 31 -- Gecko/20040804 Netscape/7.2 (ax) 8, 31 -- References: {200603201709.k2KH9SMU030393-at-ns.microscopy.com} ==============================End of - Headers==============================
Hi Barbara, There should be several responses concerning analytical labs in the archives for the listserver as this topic always becomes important when one has to build a lab. The archives can be checked at the listserver address. Also, I am sending you off line (so attachments can be included) one of my articles and checklists for designing analytical facilities. If you have any questions, please let me know. In my many years in microscopy, that has been the part that has been the most fun i.e. designing labs and I have been lucky enough to design many of them and all of them definitely represent a different challenge. Best of Luck,
Judy
Judy Murphy, PhD Microscopy Training, Imaging, and Lab Design Stockton, CA 95219 murphyjudy-at-comcast.net
b.ambrose-at-massey.ac.nz wrote:
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We are going to add a Hitachi S-2700 SEM to our lab but it is missing the instruction book and diagrams. I have a request in to Hitachi to see if they can come up with anything, but just in case, anyone have a set to share? Maybe I can copy them and get them back to you ASAP.
Thanks
Jon -- Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 4, 21 -- From jmkrupp-at-ucsc.edu Mon Mar 20 16:53:14 2006 4, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 4, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2KMrDwb026767 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 16:53:13 -0600 4, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 4, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.1/8.13.1) with ESMTP id k2KMloEZ023619 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 20 Mar 2006 14:47:58 -0800 (PST) 4, 21 -- Received: from [128.114.25.214] (account jmkrupp-at-ucsc.edu HELO [128.114.25.197]) 4, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 4, 21 -- with ESMTPA id 49810387 for microscopy-at-microscopy.com; Mon, 20 Mar 2006 14:47:49 -0800 4, 21 -- Mime-Version: 1.0 4, 21 -- Message-Id: {p06230907c044e0eacbff-at-[128.114.25.197]} 4, 21 -- Date: Mon, 20 Mar 2006 14:47:45 -0800 4, 21 -- To: microscopy-at-microscopy.com 4, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 4, 21 -- Subject: Hitachi S-2700 instructions 4, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 4, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 4, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 4, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (csucla-at-charter.net) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, March 20, 2006 at 13:40:39 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both csucla-at-charter.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: csucla-at-charter.net Name: Sandra Brodwin
Organization: Liebman & Associates
Education: Graduate College
Location: La Canada, California, USA
Title: work situation of a microscopist
Question: I am a rehabilitation consultant. I have been asked to evaluate the worksite of a microscopist who has bilateral carpel tunnel for suggestions on how to change her work situation to allow her to continue to do the work in her field. Therefore, I need to know 1) what to look for in her work station and 2) idea on how to change/adapt the work station to accommodate bilateral carpal tunnel syndrome.
You don't say whether you are talking about light microscopy or some form of electron microscopy or something else.
I would guess that it would be absolutely imperative to find out how the sufferer works with the instrument(s) in question. It sounds as if they may be using rotary controls to shift specimens around repeatedly and possibly the layout of the instrument/workstation doesn't encourage a sensible close and comfortable seating position. These are certainly problems I have heard of when someone sits at a scanning electron microscope and needs to examine lots of areas on a specimen and I would suspect that their may be similar problems with many light microscope specimen stages.
I think that the problem is that many manufacturers did not envisage operators sitting at their microscopes for 6 or 7 hours and constantly rotating the control knobs. But I suppose that it may happen in medical screening or quality control where repetitive work loads can be high.
Some manufacturers may provide more ergonomic control systems, work planning or furniture might be re-arranged. But it's difficult to suggest more without some detail.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: csucla-at-charter.net
(Posting for a colleague in Bethesda,MD)
Department of Health and Human Services
National Institutes of Health, USA
National Heart, Lung and Blood Institute
Biologist (Electron Microscopy)
The Electron Microscopy Core Facility of the Genetics and Development Biology Center, Division of Intramural Research in the National Heart, Lung and Blood Institute, is recruiting for a Biologist (Electron Microscopy). To be minimally qualified you must have a bachelor's degree in biology or related field. Experience is required in performing fixation, embedding and ultra-thin sectioning of samples for transmission electron microscopy as well as operating the electron microscope to record images. Experience with ultra-thin cryo-microtomy and immunogold labeling is highly desirable. Other desirable areas of experience include rotary shadowing, freeze fracture, operation of the scanning electron microscope and SEM specimen preparation. The successful candidate will be hired at a level and with salary compensation commensurate with previous experience and qualifications. The appointee must be a US citizen. For more information or to apply, please reference vacancy announcement number NHLBI-06-114086 and the website, WWW.USAJOBS.GOV {http://www.usajobs.gov/} {http://www.usajobs.gov/} .
The NIH is an Equal Opportunity Employer. Applications from women, minorities, and persons with disabilities are strongly encouraged. The NHLBI/NIH is a smoke-free environment.
This email and its attachments are intended for the use of the individual or entity who is the intended recipient and may contain information that is privileged, confidential and exempt from disclosure or any type of use under applicable law. If the reader of this email is not the intended recipient, or the employee, agent or representative responsible for delivering the email to the intended recipient, you are hereby notified that any dissemination, distribution, copying or other use of this email is strictly prohibited. If you have received this email in error, please reply immediately to the sender. MA
Malcolm's reply has covered the microscopes. However, modern optical and SEM microscopes invariably have a computer and mouse attached to them these days, or the user then goes back to the office PC, so CTS is a very common problem. I use mouse controlled PC's all day for microscope image capture and control, associated office and image analysis work, and at home for fun throughout the night. I also play with my young son on the Playstation (Tekken kick boxing in particular is a killer for the wrist). So naturally I get mild CTS occasionally - a clear sign to use the mouse less for a few days, switch to a racing sim, and so rest the wrist for a while.
Try using the keyboard (e.g. for shortcuts) if you have real problems. Also you can get a Cirque Smartcat laptop style large USB touchpad to replace the mouse (for about Ł60) if you have limited movement - or at least get a decent optical mouse - I prefer the MS optical Intellimouse although I disable the extra side buttons. I've never tried the Smartcat, as it will be a bit slower than a mouse, although it did receive a very good review (www.pcpro.co.uk). I have tried wrist supports (with gels etc.) but I found them very uncomfortable and stick to resting my wrist on the benchtop. A visit to my chiropractor occasionally does help me (joint manipulation and firing a little captive bolt thingy at the affected wrist bones) and for convenience I use a freeze gel rather than icepacks to reduce inflammation (so I can still use the mouse during the 'physiotherapy'). Arranging the workstation helps with neck pain but does not really do anything for my CTS, although wrist angle is supposedly important. I never find the microscope focus knob a problem at all - suppose most of ours are motorised and so have far less resistance than manual focus gears. On all our mechanical stages the manual XY stage control is on the left side, and so won't irritate the same wrist (the right hand is used for the focus).
UCl have standard health & safety handouts for workstation setup similar to http://www.afscme.org/health/faq-cts.htm but offer us no real specific advice on CTS
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {csucla-at-charter.net} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, March 21, 2006 12:25 AM
Forgot to add the link, but also have a look at
http://www.safecomputing.com
for other PC based anti-RSI ideas.
Keith ---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {csucla-at-charter.net} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, March 21, 2006 12:25 AM
Position available: Professor in Electron Microscopy and Director of the Center for Electron Nanoscopy, Technical University of Denmark Application deadline: 2 May 2006
The Technical University of Denmark (DTU) announces a position as full professor in Electron Microscopy to be filled as soon as possible. The professor will lead research in Electron Microscopy and at the same time serve as Director for our new Center for Electron Nanoscopy (CEN).
We are in the process of establishing an advanced center for electron microscopy, which will complement our strong activities within nanotechnology and materials science. The center, which is made possible by a generous donation by the A.P. Moller Foundation, will be equipped with a total of six microscopes of which two will be absolute state of the art TEM's with sub-Ĺngstrom resolution, energy loss spectroscopy, and 3D structural analysis facilities. Moreover, one of the TEM's will be modified into an environmental TEM. CEN is expected to be fully operational in the fall of 2007.
We seek a dynamic person, who will carry out advanced research in Electron Microscopy and at the same time direct activities in CEN. In the start-up phase of CEN we expect the candidate to interact with equipment manufacturers, to coordinate hiring of additional staff and to overlook the construction of a new building for CEN. When CEN is established the candidate will be responsible for the daily operation, the portfolio of research projects and the services offered, assisted by two senior scientists and several technicians. CEN will interact with a number of DTU departments within the fields related to nanotechnology including metallurgy, polymers and advanced materials, catalysis, solid state physics, micro- and nano-electronics, optics and photonics. Moreover, CEN will interact with outside partners from academia and industry. It will be the responsibility of the candidate to ensure that CEN has the capability to carry out both advanced research projects as well as general analysis. In the capacity of Director the candidate reports to a Board under the Rector of DTU.
We seek a candidate with the highest academic qualifications and demonstrated abilities in leading advanced research at an international level related to electron microscopy. We envision a background in physics or engineering with expertise in the fields of nanotechnology, materials science or chemistry and with the following qualifications:
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· Outstanding expertise in the use of electron microscopy as part of advanced research within materials science/nanotechnology
· Ability to manage and strengthen a team of scientists and technicians
· Flexibility in assuring a balance between advanced research projects and general type service jobs in the center.
· Ability to collaborate actively with many research groups in different fields and to initiate new contacts.
We expect the candidate to participate actively in teaching, development of new courses in electron microscopy as well as to supervise students both at M.Sc. and Ph.D. levels. Teaching experience at university level in an international student environment is therefore considered an advantage.
The salary and appointment terms will be negotiated in accordance with the current collective agreement for Danish University faculty members.
All interested candidates irrespective of age, gender, race, religion or ethnic background are invited to apply.
Applications should include a detailed resumé with a list of publications, a statement of teaching and research interests as well as visions and plans for the future development of the Center of Nanoscopy. Copies of key publications for assessment of the research experience should be included in the application that should be sent to Rector Lars Pallesen, Building 101 A, Technical University of Denmark, DK-2800 Lyngby, Denmark.
For more information contact Dean of Research, Prof. Kristian Stubkjaer (forskningsdekan-at-adm.dtu.dk, direct telephone +45 4525 1008).
The application with enclosures in triplicate must be received before 2 May 2006 at 12:00.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both nicol-at-semiconductor.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: nicol-at-semiconductor.com Name: Nick Aitken
Title-Subject: [Filtered] sputter coaters
Question: Hi all,
we are currently looking at replacing our old Hummer X coater. It is not producing coatings that look good enough at high magnification. We are currently looking at a couple of turbo pumped coaters by various manufacturers, but I thought I would get some external opinions. First of all our requirements: We need a good conductive coating that will not show a lot of grain at around 250 thousand times mag : repeatability is a must. We have multiple users preparing many samples and we need consistent results across the board
What manufacturers should we be looking at, and what target material would be the best for this type of use? We would be coating semiconductor devices in both topographical and cross sectional orientations.
Hi folks, CCD camera technology has moved on a lot since I bought one for our optical microscopes back in 1998. The camera still works fine, but it has always struggled with low light levels (on-chip integration being an expensive option in those days). Does anyone have any recommendations for a relatively cheap camera (+ controller/software + framegrabber) which would go on a bog standard pentium III PC, which can go to low light levels? I don't need to 'see in the dark', just do dark field optical microscopy and some macro photography with the aperture closed right down to get a good depth of field.
Many thanks in advance
Richard ________________________________________ Richard Beanland Analytical Services Bookham Inc Caswell Towcester Northants NN12 8EQ United Kingdom Tel. +44 1327 356362 Fax. +44 1327 356775 http://www.bookham.com ________________________________________
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==============================Original Headers============================== 6, 31 -- From richard.beanland-at-bookham.com Tue Mar 21 07:58:23 2006 6, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 6, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2LDwMjn019156 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 07:58:23 -0600 6, 31 -- X-VirusChecked: Checked 6, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 6, 31 -- X-Msg-Ref: server-14.tower-78.messagelabs.com!1142949501!43371919!1 6, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 6, 31 -- X-Originating-IP: [213.249.209.179] 6, 31 -- Received: (qmail 5026 invoked from network); 21 Mar 2006 13:58:21 -0000 6, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 6, 31 -- by server-14.tower-78.messagelabs.com with SMTP; 21 Mar 2006 13:58:21 -0000 6, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 6, 31 -- Tue, 21 Mar 2006 13:58:20 +0000 6, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 31 -- Content-class: urn:content-classes:message 6, 31 -- MIME-Version: 1.0 6, 31 -- Content-Type: text/plain; 6, 31 -- charset="us-ascii" 6, 31 -- Subject: LM: integrating digital camera 6, 31 -- Date: Tue, 21 Mar 2006 13:58:19 -0000 6, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E047CED-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 6, 31 -- X-MS-Has-Attach: 6, 31 -- X-MS-TNEF-Correlator: 6, 31 -- Thread-Topic: LM: integrating digital camera 6, 31 -- Thread-Index: AcZM74IXGTGLQD+wSZCV9dT4DMyAFw== 6, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 6, 31 -- To: {microscopy-at-microscopy.com} 6, 31 -- X-OriginalArrivalTime: 21 Mar 2006 13:58:20.0696 (UTC) FILETIME=[83189980:01C64CEF] 6, 31 -- Content-Transfer-Encoding: 8bit 6, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2LDwMjn019156 ==============================End of - Headers==============================
I am currently using one to observe electron diffraction. However, this camera only has VGA resolution (640 x 480 pixels). It has a USB interface, so you don't need a framegrabber. And the software it comes with is worth the $100 alone - you can use it to change the integration time of the CMOS sensor for low light levels.
I've been very happy with this little product, but you might look at Orion's other products if you want a higher-resolution camera.
Ben McMorran Research Associate, Atom Optics Group Department of Physics University of Arizona 1118 E 4th St Tucson, AZ 85721
ph. 520-621-2688
Quoting richard.beanland-at-bookham.com:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi folks, } CCD camera technology has moved on a lot since I bought one for } our optical microscopes back in 1998. The camera still works fine, but } it has always struggled with low light levels (on-chip integration being } an expensive option in those days). Does anyone have any } recommendations for a relatively cheap camera (+ controller/software + } framegrabber) which would go on a bog standard pentium III PC, which can } go to low light levels? I don't need to 'see in the dark', just do dark } field optical microscopy and some macro photography with the aperture } closed right down to get a good depth of field. } } Many thanks in advance } } Richard } ________________________________________ } Richard Beanland } Analytical Services } Bookham Inc } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } } ======================================================================= } This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly prohibited. } No part of this message can be considered a request for goods or } services. } ======================================================================= } } } ==============================Original Headers============================== } 6, 31 -- From richard.beanland-at-bookham.com Tue Mar 21 07:58:23 2006 } 6, 31 -- Received: from mail78.messagelabs.com } (mail78.messagelabs.com [195.245.230.131]) } 6, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2LDwMjn019156 } 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 07:58:23 -0600 } 6, 31 -- X-VirusChecked: Checked } 6, 31 -- X-Env-Sender: richard.beanland-at-bookham.com } 6, 31 -- X-Msg-Ref: server-14.tower-78.messagelabs.com!1142949501!43371919!1 } 6, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- } 6, 31 -- X-Originating-IP: [213.249.209.179] } 6, 31 -- Received: (qmail 5026 invoked from network); 21 Mar 2006 } 13:58:21 -0000 } 6, 31 -- Received: from unknown (HELO } cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 6, 31 -- by server-14.tower-78.messagelabs.com with SMTP; 21 Mar } 2006 13:58:21 -0000 } 6, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI } ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with } Microsoft SMTPSVC(6.0.3790.211); } 6, 31 -- Tue, 21 Mar 2006 13:58:20 +0000 } 6, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 6, 31 -- Content-class: urn:content-classes:message } 6, 31 -- MIME-Version: 1.0 } 6, 31 -- Content-Type: text/plain; } 6, 31 -- charset="us-ascii" } 6, 31 -- Subject: LM: integrating digital camera } 6, 31 -- Date: Tue, 21 Mar 2006 13:58:19 -0000 } 6, 31 -- Message-ID: } {9645D3E33E4C6548B12A7B25F611533E047CED-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} } 6, 31 -- X-MS-Has-Attach: } 6, 31 -- X-MS-TNEF-Correlator: } 6, 31 -- Thread-Topic: LM: integrating digital camera } 6, 31 -- Thread-Index: AcZM74IXGTGLQD+wSZCV9dT4DMyAFw== } 6, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 6, 31 -- To: {microscopy-at-microscopy.com} } 6, 31 -- X-OriginalArrivalTime: 21 Mar 2006 13:58:20.0696 (UTC) } FILETIME=[83189980:01C64CEF] } 6, 31 -- Content-Transfer-Encoding: 8bit } 6, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k2LDwMjn019156 } ==============================End of - Headers==============================
==============================Original Headers============================== 13, 27 -- From mcmorran-at-physics.arizona.edu Tue Mar 21 09:19:56 2006 13, 27 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 13, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2LFJt0s029926 13, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 09:19:55 -0600 13, 27 -- Received: from localhost (gimli.email.arizona.edu [10.0.0.223]) 13, 27 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 564C0D415A7 13, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 08:19:55 -0700 (MST) 13, 27 -- Received: from localhost (treebeard.email.arizona.edu [10.0.0.213]) 13, 27 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9324BD50DCC 13, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 08:19:53 -0700 (MST) 13, 27 -- Received: from dhcp029.pas.arizona.edu (dhcp029.pas.arizona.edu 13, 27 -- [150.135.51.31]) by www.email.arizona.edu (Horde) with HTTP for 13, 27 -- {mcmorran-at-email.arizona.edu} ; Tue, 21 Mar 2006 08:19:53 -0700 13, 27 -- Message-ID: {20060321081953.2ssjggcg4g88s44o-at-www.email.arizona.edu} 13, 27 -- X-Priority: 3 (Normal) 13, 27 -- Date: Tue, 21 Mar 2006 08:19:53 -0700 13, 27 -- From: Ben McMorran {mcmorran-at-physics.arizona.edu} 13, 27 -- To: Microscopy-at-microscopy.com 13, 27 -- Subject: Re: [Microscopy] LM: integrating digital camera 13, 27 -- References: {200603211401.k2LE1kmh023763-at-ns.microscopy.com} 13, 27 -- In-Reply-To: {200603211401.k2LE1kmh023763-at-ns.microscopy.com} 13, 27 -- MIME-Version: 1.0 13, 27 -- Content-Type: text/plain; charset="ISO-8859-1"; format="flowed" 13, 27 -- Content-Disposition: inline 13, 27 -- Content-Transfer-Encoding: 7bit 13, 27 -- User-Agent: Internet Messaging Program (IMP) 4.0-cvs 13, 27 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Sputter coaters will often take targets made of different metals and this is what primarily determines fineness of the "grain", along with sputtering current, sputtering atmosphere, and coating times. We currently have targets made of platinum, chromium, gold, tantalum, titanium, and aluminum. For SEM conductive coating, we generally use platinum, which is much finer-grained than gold or gold/palladium. Finer yet is chromium, but that has the disadvantage of oxidizing rapidly to form an insulating layer, which almost makes it a "one-viewing" type of coating, unless the sample is stored under a good vacuum. Also, oxidizing metals require that the sputter coater be able to etch the oxide layer off the target before the actual coating cycle.
Iridium and osmium are also fine-grained coatings, but I have no personal experience with these metals. Osmium coating requires a special instrument, which is supplied by SPI Supplies and maybe others.
If you have multiple users, as we do, repeatability is important, but so is flexibility. We are constantly changing coating times and sputtering currents, depending on sample, desired magnification, and other factors. The ability to take targets of different metals is important----some require different sputtering parameters. Also, if your facility is like ours, word might get out to the electrical engineering folks that a coater is available to put down repeatable layers of metals on various substrates for making circuits and other "dark side" wizardry. If so, be prepared for some exotic requests.
Finally, if you do end up coating with a variety of metals, you can save tons of money by buying custom targets from 3rd party suppliers, such as Abe Dayani of Refining Systems, Inc. and possibly others. (No financial interest, etc., but just a satisfied customer.) Such suppliers can provide targets in almost any metal, in any size, and in any purity needed, at substantial savings over OEM prices.
Turbo coaters are definitely nice and I imagine that any coater in that price range would also provide the ability to play with coating parameters for most applications. I also recommend a rotating, tiltable specimen stage.
Hope some of this helps.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: nicol-at-semiconductor.com [mailto:nicol-at-semiconductor.com] Sent: Tuesday, March 21, 2006 7:33 AM To: Tindall, Randy D.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both nicol-at-semiconductor.com as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: nicol-at-semiconductor.com Name: Nick Aitken
Title-Subject: [Filtered] sputter coaters
Question: Hi all,
we are currently looking at replacing our old Hummer X coater. It is not producing coatings that look good enough at high magnification. We are currently looking at a couple of turbo pumped coaters by various manufacturers, but I thought I would get some external opinions. First of all our requirements: We need a good conductive coating that will not show a lot of grain at around 250 thousand times mag : repeatability is a must. We have multiple users preparing many samples and we need consistent results across the board
What manufacturers should we be looking at, and what target material would be the best for this type of use? We would be coating semiconductor devices in both topographical and cross sectional orientations.
Cr gives one of the best coatings, if not the best, for high resolution. (I highly suspect that a well known fellow from SPI will probably disagree with that statement, again.) However, it oxidizes in short time and the sample should not be exposed to air for prolonged periods of time. We have introduced the SampleSaver(TM) Storage Container system that will solve that problem. The next target materials that people use with good success is Ir and W. These also give a fine grain size and are less susceptible to oxidation. It is important that your coating system gives a very thin, but uniform coating that conforms to the surface. This basically allows the charge to be drained from the surface. With these thin, high resolution coatings, it is also important for insulating materials to keep the accelerating voltage down so that the beam is not penetrating deep into the samples and causing the charge to be trapped well below the coating where it is unable to be dissipated to ground.
Since your application is with semiconductor materials, the ability to have an etch gun is a definite plus. With our system, the etch gun can be used to low angle polish as well as etch at higher angles. The higher angle will give a differential sputter etch that can enhance contrast between different materials as well as etch grain structures in your coatings.
Disclaimer: South Bay Technology, Inc. manufactures and sells the IBS/e system and the SampleSaver(TM) Storage Container.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: nicol-at-semiconductor.com [mailto:nicol-at-semiconductor.com] Sent: Tuesday, March 21, 2006 5:35 AM To: Walck-at-SouthBayTech.com
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Email: nicol-at-semiconductor.com Name: Nick Aitken
Title-Subject: [Filtered] sputter coaters
Question: Hi all,
we are currently looking at replacing our old Hummer X coater. It is not producing coatings that look good enough at high magnification. We are currently looking at a couple of turbo pumped coaters by various manufacturers, but I thought I would get some external opinions. First of all our requirements: We need a good conductive coating that will not show a lot of grain at around 250 thousand times mag : repeatability is a must. We have multiple users preparing many samples and we need consistent results across the board
What manufacturers should we be looking at, and what target material would be the best for this type of use? We would be coating semiconductor devices in both topographical and cross sectional orientations.
I lost my second Anatech Hummer II a couple years ago did a similar search for replacements. I wound up with a Denton Desk II. It worked very well for about a year and then failed to fire or keep a steady plasma. Denton replaced the HV board on a trial swap basis but it made no difference. So something else was going on. However, as I was moving to smaller node sizes, HC contamination was becoming a real nusance. So this moved me to search for a turbo coater for finer grain at high mag.
I wound up getting a Denton Desk IV TSC with tilt and rotation. My major modification that they accommodated was the replacement of the oil diaphram pump with an Edwards XDS5 dry scroll pump. So the forepump is external from the desk-top coater. Total prep time is longer since the scroll pump does not pump as fast as an oil pump. But the results are excellent. If the TSC circuitry would support it, an XDS10 would likely be faster. Since my SEM is totally dry pumped, the whole prep scenario results in as close to zero HC contamination as possible.
The TSC uses 6cm diameter targets. Or you could use 5.75cm. The material to be used is tricky. Au produces a web or mesh coating and is not at all good for high mag. Au/Pd is good as is Pd. Better noble metals such as Pt and Ir are even better. But here is the kicker. If you do EDS, suppose you coat with Pt and the barrier layer is Pt. If you do quant in the barrier layer, it will be wrong due to the additional coating. W plugs are also an issue since its L beta is close to L alpha of Pt. I suppose my point is to not use a target material that is likely to be part of the IC. The other consideration is peak pileup at low eV. I wound up choosing Ir.
Protocol is:
put specimen in coater and close lid start rough pump wait until vac is about 322mT start turbo after Turbo -at- Speed is indicated, check terminal vacuum. Should be about 4-5mT open gas (Ar) adjust needle valve for vacuum of 15mT-20mT (depends on what final results you want) go to timed sputter set Rotate=20% (if you want rotation) set Power=60% (typically results in 12-15mA) set Time=60 seconds (adjust for your application) when done, turn off turbo (HV will already have gone off) open needle valve wide open (this increases friction at the turbo bearings to slow it down) When you hear the turbo running down to very slow, stop fore pump wait a few seconds and open the lid
This process takes longer than with the oil fore pump as I said. But there is no HC contamination as-prepared. As the specimen is left outside of the SEM and not under vacuum, there will be contamination and you will get the HC scan "burns." If you use the specimen again, stick it under a high intensity UV lamp for about two hours and see if that clears the HC.
Au/Pd, Pt, Ir range in price from about $300 to $1,000, in that order. However, the Ir target is quite thick. The others are around .008". Thickness obviously affects the price as does material and size. Abe offers targets in several thicknesses. The targets are not 99.999% pure. Each target will have a different purity and its own set of trace elements. So far, the worst target purity is 99.5% for the Pd and Ir. Trace elements are Mn, Si, Cu, Fe, P, S. But a 100-200A film is not likely to show up with such small trace values. The 0.5% trace is divided up amongst several trace elements--not just one.
Disclaimer: I have no financial interest in Denton or Refining Systems other than to hope they stay around to offer good products and good service.
Happy coating, gary g.
At 05:34 AM 3/21/2006, you wrote:
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==============================Original Headers============================== 19, 20 -- From gary-at-gaugler.com Tue Mar 21 12:25:23 2006 19, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 19, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2LIPNM1029314 19, 20 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 12:25:23 -0600 19, 20 -- Received: (qmail 2136 invoked from network); 21 Mar 2006 10:25:22 -0800 19, 20 -- Received: by simscan 1.1.0 ppid: 2132, pid: 2133, t: 0.2250s 19, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 19, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 19, 20 -- by qsmtp3 with SMTP; 21 Mar 2006 10:25:22 -0800 19, 20 -- Message-Id: {6.2.3.4.2.20060321093615.023c3770-at-mail.calweb.com} 19, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 19, 20 -- Date: Tue, 21 Mar 2006 10:25:25 -0800 19, 20 -- To: nicol-at-semiconductor.com 19, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 19, 20 -- Subject: Re: [Microscopy] viaWWW: sputter coaters 19, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 19, 20 -- In-Reply-To: {200603211334.k2LDY6II012454-at-ns.microscopy.com} 19, 20 -- References: {200603211334.k2LDY6II012454-at-ns.microscopy.com} 19, 20 -- Mime-Version: 1.0 19, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
As a consultant I am often asked to appraise a laboratory and its staff. X-from this, with my interests in Quality in Electron Microscopy, I am involved with a paper related to the running of EM units and to training levels. The referees are happy with the paper but would like to see some results from a range of different units. They are interested in which are areas of action in a laboratory and which are the areas that do not seem to be active.
We have put together an EM Unit Survey on an Excel file where through the totting up of points we will be able to see where laboratories stand. Knowing how political these results could be I have arranged for a kart race colleague to collate them for us and to discarding their email source once the Excel file has been collected. In this way we will be able to obtain information but neither the authors of the paper nor others will know the source of the data. The Excel file is available on the Protrain web site under the Hints and Tips section for those who wish to help us.
If you would be able to help us it requires selecting your response to the Excel file and forwarding it to info-at-whiltonmillkartclub.co.uk .
Many thanks if you are able to help. If you would like to receive the data when collated please mail me separately.
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7802 966067 Web www.emcourses.com
==============================Original Headers============================== 7, 29 -- From protrain-at-emcourses.com Tue Mar 21 13:25:34 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 (8.12.11/8.12.8) with ESMTP id k2LJPXJI007875 7, 29 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 13:25:34 -0600 7, 29 -- Received: from [62.69.65.167] (helo=smtp-d.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 1FLmU1-0005wD-89 7, 29 -- for microscopy-at-microscopy.com; Tue, 21 Mar 2006 19:25:33 +0000 7, 29 -- Received: (qmail 10644 invoked from network); 21 Mar 2006 19:25:26 -0000 7, 29 -- Received: from unknown (HELO advent) (212.248.147.118) 7, 29 -- by 0 with SMTP; 21 Mar 2006 19:25:26 -0000 7, 29 -- Message-ID: {003301c64d1d$805e1e80$7693f8d4-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: EM Unit Survey 7, 29 -- Date: Tue, 21 Mar 2006 19:16:21 -0000 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==============================
Just a reminder, the Microscopy Listserver rules do not permit the posting of survey's without first receiving explicit permission.
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This Question was submitted to Ask-A-Microscopist by (jminarcik-at-sbcglobal.net) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, March 21, 2006 at 15:49:27 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both jminarcik-at-sbcglobal.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: jminarcik-at-sbcglobal.net Name: John Minarcik
Organization: Hard Knox
Education: Graduate College
Location: City, State, Country
Title: Plastic embedded super high quality H&E histo slides
Question: I need a set of super high quality plastic-embedded general H&E slides covering all types of tissues/organs.
We are looking for a disposable biopsy punch to use for cutting samples from flower petals and other plant material. I have found references to the Harris Uni-core punch and to the Miltex punches (used by our VET school).
I would appreciate hearing from anyone who has used either type (or has a recommendation for another type) and a source for ordering them. We would like to use them more than once if possible. I am sure number of uses depends on type of sample.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 6, 21 -- From dsherman-at-purdue.edu Tue Mar 21 20:43:40 2006 6, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 6, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M2hdZC011497 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 20:43:39 -0600 6, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Tue, 21 Mar 2006 21:43:38 -0500 6, 21 -- Received: from 12.222.30.139 ([12.222.30.139]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 6, 21 -- Wed, 22 Mar 2006 02:33:55 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 6, 21 -- Date: Tue, 21 Mar 2006 21:33:54 -0500 6, 21 -- Subject: Biopsy punch 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C04621C2.12C7%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Biopsy punch 6, 21 -- Thread-Index: AcZNWQ/ZTi8fSblMEdqUkwAKlcoUxg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 22 Mar 2006 02:43:38.0969 (UTC) FILETIME=[6C850490:01C64D5A] ==============================End of - Headers==============================
I am very happy to report that the Facility Organization and Management FIG is sponsoring a special pre-meeting workshop in Chicago on Saturday July 29 from 1-5pm prior to the start of M&M 2006. Information is posted on the MSA web site under annual meeting-2006. Select Pre-meeting events.
http://mm2006.microscopy.org/
I am really excited about the workshop as it will cover topics quite different than those covered in past sessions on Core Facility Management. The presenters are all professionals from the business and management areas, not microscopists. Having a workshop run by non-microscopists but professionals in management can help us think łout of the box˛ as we develop new strategies for helping keep our facilities viable.
Facility Operations and Management FIG Workshop
Saturday, July 29, 1:00-5:00pm (prior to M&M 2006) Holiday Inn City Center Hotel, Chicago, IL Registration fee: $40 Space is limited. You do not have to be a FOM FIG member to attend the workshop. However, you must download the form from the MSA meeting website and submit it with your registration fee.
The Workshop title is: New Approaches to Marketing, Managing, and Money for Maintaining a Core Facility (4Mąs)
Topics: 1) How to Make a Business Plan for short and Long-term Facility Maintenance and Growth. Donald Blewett, Associate Director, Burton Morgan Center for Entrepreneurship, Purdue University
3) Marketing a Facility to Increase and Maintain a User Base (and maintain the support of the upper administration). Dr. George Adams, Research Development manager, Birck nanotechnology Center, Purdue University
4) Developing a financial plan for the long-term łcare and feeding˛ of Major Equipment. Charlene Sullivan, Professor, Krannert School of Management, Purdue University Note: extensive research and information from many microscopy facilities have provided the raw data for the analysis needed to develop this plan. The results will be presented at the workshop.
Hope to see many of you there.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 16, 22 -- From dsherman-at-purdue.edu Tue Mar 21 21:04:59 2006 16, 22 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 16, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M34w3v015807 16, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 21:04:58 -0600 16, 22 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 16, 22 -- Tue, 21 Mar 2006 22:04:58 -0500 16, 22 -- Received: from 12.222.30.139 ([12.222.30.139]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 16, 22 -- Wed, 22 Mar 2006 03:04:58 +0000 16, 22 -- User-Agent: Microsoft-Entourage/11.2.1.051004 16, 22 -- Date: Tue, 21 Mar 2006 22:04:57 -0500 16, 22 -- Subject: Facility Management Workshop-M&M2006 16, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 16, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 16, 22 -- Message-ID: {C0462909.12D2%dsherman-at-purdue.edu} 16, 22 -- Thread-Topic: Facility Management Workshop-M&M2006 16, 22 -- Thread-Index: AcZNXWZJpKSgCrlQEdqUkwAKlcoUxg== 16, 22 -- Mime-version: 1.0 16, 22 -- Content-type: text/plain; 16, 22 -- charset="ISO-8859-1" 16, 22 -- X-OriginalArrivalTime: 22 Mar 2006 03:04:58.0438 (UTC) FILETIME=[67247E60:01C64D5D] 16, 22 -- Content-Transfer-Encoding: 8bit 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2M34w3v015807 ==============================End of - Headers==============================
Well, I am probably not the guy who Scott referred to but I think that one can't really talk sensibly about coating for the SEM without talking about beam kV. As some of you may remember, when the first immersion-lens FE scopes arrived, I was one of the first to champion 1.5 kV for HR SEM of biological specimens. At low kV, you don't need much conductivity because the SE coef if higher and you don't need so much beam current because the contrast is higher to, but you do still usually need some coating (for surface contrast, even if nothing else.).
We may have made our Cr coats wrong but David Joy once opined that one can't have a "few-nw-thick" film of Cr and he looked at a lot of them in in TEM with energy loss to detect the oxide. Cr will oxidize in a flash, even in microscope vacuum (remember, 10*-6 torr is one monolayer/second and a lot of that monolayer is likely to be water vapor, which becomes oxygen if the beam hits it. You may have a cold trap but, as the "ice" that forms on it is amorphous, it has a much higher vapor pressure than you think.)
So we could never get Cr to work. Specimens charged and the contrast was not great. Also, it is important to remember that, as Cr-oxide is about 3x less dense than Cr metal, the geometric thickness is about 3x thicker than that indicated by the crystal monitor on your coater.
We could not see shall surface structures.
Instead, we used ion-beam sputtered Pt. It does form 1-2 nm crystals (about 3-5 atoms on a side) about 1-2 nm apart when put on at about 1 mn average thickness. You see this pattern when you look at the specimen at 5kV where the beam is smaller and Z-contrast makes them visible, even in SE, but the surface contrast is rudimentary.
Fortunately, these crystals are usually not visible at 1.5kV and they certainly keep the charging under control and provide some surface contrast. We all like "resolution" but remember that few specimen preparation methods preserve structure below 3nm anyway.
And Pt films are MUCH less reactive than Cr.
Not to push it but metals conduct BECAUSE of their crystalline structure. Amorphous metals are much less conductive. Metal oxides less still. Cr films have a conductivity similar to that of carbon films.
Many people use Cr at high kV but I will leave it to others to discuss the pro's and con's.
Cheers,
Jim Pawley (not only a confocalite!)
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-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 10-22, 2006, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2006 "If it ain't diffraction, it must be statistics." Anon.
==============================Original Headers============================== 15, 27 -- From jbpawley-at-wisc.edu Tue Mar 21 23:50:00 2006 15, 27 -- Received: from smtp5.wiscmail.wisc.edu (heimdall.doit.wisc.edu [144.92.197.159]) 15, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M5nxK1001783 15, 27 -- for {Microscopy-at-Microscopy.Com} ; Tue, 21 Mar 2006 23:49:59 -0600 15, 27 -- Received: from avs-daemon.smtp5.wiscmail.wisc.edu by smtp5.wiscmail.wisc.edu 15, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.08 (built Sep 22 2005)) 15, 27 -- id {0IWI00D17LJBAZ-at-smtp5.wiscmail.wisc.edu} for Microscopy-at-Microscopy.Com; 15, 27 -- Tue, 21 Mar 2006 23:49:59 -0600 (CST) 15, 27 -- Received: from [172.16.1.41] ([144.92.238.207]) by smtp5.wiscmail.wisc.edu 15, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.08 (built Sep 22 2005)) 15, 27 -- with ESMTPSA id {0IWI00K1XLJ87C-at-smtp5.wiscmail.wisc.edu} ; Tue, 15, 27 -- 21 Mar 2006 23:49:57 -0600 (CST) 15, 27 -- Date: Tue, 21 Mar 2006 23:49:52 -0600 15, 27 -- From: James Pawley {jbpawley-at-wisc.edu} 15, 27 -- Subject: [Microscopy] RE: viaWWW: sputter coaters 15, 27 -- In-reply-to: {200603211721.k2LHLHHZ027169-at-ns.microscopy.com} 15, 27 -- X-Sender: jbpawley-at-wiscmail.wisc.edu 15, 27 -- To: walck-at-southbaytech.com, 15, 27 -- "ListServer-at-MSA.Microscopy.Com" {Microscopy-at-Microscopy.Com} 15, 27 -- Message-id: {p06110417c0468f14149e-at-[172.16.1.41]} 15, 27 -- MIME-version: 1.0 15, 27 -- Content-type: text/plain; format=flowed; charset=us-ascii 15, 27 -- Content-transfer-encoding: 7BIT 15, 27 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=[144.92.238.207] 15, 27 -- X-Spam-PmxInfo: Server=avs-1, Version=5.1.2.240295, Antispam-Engine: 2.3.0.1, 15, 27 -- Antispam-Data: 2006.3.21.213107, SenderIP=[144.92.238.207] 15, 27 -- References: {200603211721.k2LHLHHZ027169-at-ns.microscopy.com} ==============================End of - Headers==============================
Please note... the speaker topics were miss numbered (thank WORD for auto correcting again!). There are only 3 speakers. Sorry for clogging your in box twice with the same message.
Debby
} From: {dsherman-at-purdue.edu} } Reply-To: {dsherman-at-purdue.edu} } Date: Tue, 21 Mar 2006 21:38:33 -0600 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] Facility Management Workshop-M&M2006 } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } List members, } } I am very happy to report that the Facility Organization and Management FIG } is sponsoring a special pre-meeting workshop in Chicago on Saturday July 29 } from 1-5pm prior to the start of M&M 2006. Information is posted on the } MSA web site under annual meeting-2006. Select Pre-meeting events. } } http://mm2006.microscopy.org/ } } I am really excited about the workshop as it will cover topics quite } different than those covered in past sessions on Core Facility Management. } The presenters are all professionals from the business and management areas, } not microscopists. Having a workshop run by non-microscopists but } professionals in management can help us think łout of the box˛ as we develop } new strategies for helping keep our facilities viable. } } } Facility Operations and Management FIG Workshop } } Saturday, July 29, 1:00-5:00pm (prior to M&M 2006) } Holiday Inn City Center Hotel, Chicago, IL } Registration fee: $40 Space is limited. } You do not have to be a FOM FIG member to attend the workshop. However, you } must download the form from the MSA meeting website and submit it with your } registration fee. } } } The Workshop title is: } New Approaches to Marketing, Managing, and Money for Maintaining a } Core Facility (4Mąs) } } Topics: } 1) How to Make a Business Plan for short and Long-term Facility } Maintenance and Growth. } Donald Blewett, Associate Director, Burton Morgan Center for } Entrepreneurship, Purdue University } } 2) Marketing a Facility to Increase and Maintain a User Base (and } maintain the support of the upper administration). } Dr. George Adams, Research Development manager, Birck nanotechnology Center, } Purdue University } } 3) Developing a financial plan for the long-term łcare and feeding˛ of } Major Equipment. } Charlene Sullivan, Professor, Krannert School of Management, Purdue } University } Note: extensive research and information from many microscopy facilities } have provided the raw data for the analysis needed to develop this plan. The } results will be presented at the workshop. } } Hope to see many of you there. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www.agriculture.purdue.edu/microscopy } } } } ==============================Original Headers============================== } 16, 22 -- From dsherman-at-purdue.edu Tue Mar 21 21:04:59 2006 } 16, 22 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu } [128.210.63.223]) } 16, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2M34w3v015807 } 16, 22 -- for {microscopy-at-microscopy.com} ; Tue, 21 Mar 2006 21:04:58 -0600 } 16, 22 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by } exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 16, 22 -- Tue, 21 Mar 2006 22:04:58 -0500 } 16, 22 -- Received: from 12.222.30.139 ([12.222.30.139]) by EXCH02.purdue.lcl } ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu } ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; } 16, 22 -- Wed, 22 Mar 2006 03:04:58 +0000 } 16, 22 -- User-Agent: Microsoft-Entourage/11.2.1.051004 } 16, 22 -- Date: Tue, 21 Mar 2006 22:04:57 -0500 } 16, 22 -- Subject: Facility Management Workshop-M&M2006 } 16, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} } 16, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 16, 22 -- Message-ID: {C0462909.12D2%dsherman-at-purdue.edu} } 16, 22 -- Thread-Topic: Facility Management Workshop-M&M2006 } 16, 22 -- Thread-Index: AcZNXWZJpKSgCrlQEdqUkwAKlcoUxg== } 16, 22 -- Mime-version: 1.0 } 16, 22 -- Content-type: text/plain; } 16, 22 -- charset="ISO-8859-1" } 16, 22 -- X-OriginalArrivalTime: 22 Mar 2006 03:04:58.0438 (UTC) } FILETIME=[67247E60:01C64D5D] } 16, 22 -- Content-Transfer-Encoding: 8bit } 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k2M34w3v015807 } ==============================End of - Headers==============================
==============================Original Headers============================== 6, 23 -- From dsherman-at-purdue.edu Wed Mar 22 08:19:52 2006 6, 23 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2MEJp1q027971 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 08:19:51 -0600 6, 23 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 23 -- Wed, 22 Mar 2006 09:19:51 -0500 6, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 6, 23 -- Wed, 22 Mar 2006 14:19:51 +0000 6, 23 -- User-Agent: Microsoft-Entourage/11.2.1.051004 6, 23 -- Date: Wed, 22 Mar 2006 09:19:51 -0500 6, 23 -- Subject: Correction- Facility Management Workshop-M&M2006 6, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 23 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 23 -- Message-ID: {C046C737.E4EE%dsherman-at-purdue.edu} 6, 23 -- Thread-Topic: Correction- Facility Management Workshop-M&M2006 6, 23 -- Thread-Index: AcZNu66X7Q7MormuEdq0UwARJN08Mg== 6, 23 -- In-Reply-To: {200603220338.k2M3cXN7024849-at-ns.microscopy.com} 6, 23 -- Mime-version: 1.0 6, 23 -- Content-type: text/plain; 6, 23 -- charset="ISO-8859-1" 6, 23 -- X-OriginalArrivalTime: 22 Mar 2006 14:19:51.0360 (UTC) FILETIME=[AECE0C00:01C64DBB] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2MEJp1q027971 ==============================End of - Headers==============================
Are there tight junctions (zonula occludens) in keratinized stratified squamous epithelia such as skin? Or do the lamellar granules (membrane coating granules) take care of all the sealing needed? As a corollary- are there tight junctions in non-keratinizing stratified epithelia such as esophagus? thanks for some basic histo I should know! tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have tried to give away a bunch of photo paper we don't use any more with no luck.
It is a little old, a couple of years, but it has been in a refrigerator and should be OK. It's not getting any younger and I don't think we are ever going to use it.
It is Kodak Kodabrome II RC in various contrast grades, 1 -5.
If you can use it, contact me and we can work out some way to get it to you.
Jon
-- Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 7, 21 -- From jmkrupp-at-ucsc.edu Wed Mar 22 12:01:36 2006 7, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2MI1XTt010362 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 12:01:33 -0600 7, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 7, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.1/8.13.1) with ESMTP id k2MHlobo017450 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 09:47:51 -0800 (PST) 7, 21 -- Received: from [128.114.25.214] (account jmkrupp-at-ucsc.edu [128.114.25.214] verified) 7, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 7, 21 -- with ESMTPA id 50032116 for microscopy-at-microscopy.com; Wed, 22 Mar 2006 09:47:50 -0800 7, 21 -- Mime-Version: 1.0 7, 21 -- Message-Id: {p06230902c0473d8062cf-at-[128.114.25.214]} 7, 21 -- Date: Wed, 22 Mar 2006 09:47:49 -0800 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 7, 21 -- Subject: Surplus photo paper 7, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 7, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 7, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 7, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (alvarobq-at-fcien.edu.uy) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, March 22, 2006 at 11:08:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both alvarobq-at-fcien.edu.uy as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I`ll very appreciate if you can send me the e-mail address of a Quetol 651 supplier. I need to know more about this resin to compare to Spurr. If every one have some experience, please send me any advise. Thank you very much.
This Question was submitted to Ask-A-Microscopist by (alvarobq-at-fcien.edu.uy) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, March 22, 2006 at 11:08:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both alvarobq-at-fcien.edu.uy as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I`ll very appreciate if you can send me the e-mail address of a Quetol 651 supplier. I need to know more about this resin to compare to Spurr. If every one have some experience, please send me any advise. Thank you very much.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rpetrova-at-mail.ucf.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpetrova-at-mail.ucf.edu Name: Rumy
Organization: UCF
Title-Subject: [Filtered] sample prep
Question: Does anyone know what is the best way to polish CdZnO ? Would the wedge polishing work,using diamond lapping film?
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both v_bleu_knight-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Knives for Ultrathin sectioning of bone or calcium crystals
Question: Hello List,
Does anyone have experience creating ultrathin sections of tissues that have bone or calcium crystals in them? If so, what kind of diamond knife are you using? I am reluctant to use my standard 45 degree diamond knife because I feel that it would dull very quickly. Thank you in advance for your response.
Sincerely, Bleu Knight PhD Candidate New Mexico State University
I have tried to give away a bunch of photo paper we don't use any more with no luck.
It is a little old, a couple of years, but it has been in a refrigerator and should be OK. It's not getting any younger and I don't think we are ever going to use it.
It is Kodak Kodabrome II RC in various contrast grades, 1 -5.
If you can use it, contact me and we can work out some way to get it to you.
Jon
-- Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 7, 21 -- From jmkrupp-at-ucsc.edu Wed Mar 22 18:39:50 2006 7, 21 -- Received: from smtp-prod-mx3.ucsc.edu (smtp-prod-mx3.ucsc.edu [128.114.125.45]) 7, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2MI1XTt010362 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 12:01:33 -0600 7, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 7, 21 -- by smtp-prod-mx3.ucsc.edu (8.13.1/8.13.1) with ESMTP id k2MHlobo017450 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Mar 2006 09:47:51 -0800 (PST) 7, 21 -- Received: from [128.114.25.214] (account jmkrupp-at-ucsc.edu [128.114.25.214] verified) 7, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 7, 21 -- with ESMTPA id 50032116 for microscopy-at-microscopy.com; Wed, 22 Mar 2006 09:47:50 -0800 7, 21 -- Mime-Version: 1.0 7, 21 -- Message-Id: {p06230902c0473d8062cf-at-[128.114.25.214]} 7, 21 -- Date: Wed, 22 Mar 2006 09:47:49 -0800 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 7, 21 -- Subject: Surplus photo paper 7, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 7, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 7, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 7, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
Are there tight junctions (zonula occludens) in keratinized stratified squamous epithelia such as skin? Or do the lamellar granules (membrane coating granules) take care of all the sealing needed? As a corollary- are there tight junctions in non-keratinizing stratified epithelia such as esophagus? thanks for some basic histo I should know! tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I'm looking into writing javascript for reading this SEM data (e.g., magnification) that is embedded in the TIFF files in a non-standard way. I expect the javascript to work within Photoshop, and I'd certainly be interested if anyone has already done this and can provide an example (although I am sure my SEM data is formatted differently). (BTW, I already aware I can open these files with a text editor and simply extract the text ... But that's not very elegant ...)
The 2nd issue is what to do with the data once retrieved? I could simply write it to a text file, but it would be better to write it back to the TIFF in a standard way. For example, to put it all in the EXIF "comments" field ... Or even better, to put it where it should be put ... Except I can find no information as to Microscopists, as a group, asking that standard EXIF fields be designated and standardized (e.g., the "microns per pixel" field).
When we upgraded the computer and software for our JEOL 5600, I was so disappointed that the information that can be embedded into images was still in the 1970s style, taking up a significant portion of the bottom of the image, and not adjustable in any way, that I bit the bullet and wrote my own program to add a strip at the bottom of the images with information from the SEM data text file. This extended the dimensions of the 1280x960 image to 1280x1024, but none of the image area is taken up by those blocky looking, primitive characters. You can see examples from the page I recently posted from the "mystery object/starch grain" thread a month or so back:
http://www.mta.ca/dmf/download/ehrman/mystery.htm
In writing this, I was also wondering how "standard" some of this information is across instrument models and manufacturers? If people are interested, send me an email with one of your text files, along with the instrument make and model, I can do some comparisons. Along those lines, would anybody be interested in an application that would write this information to their images? If their is some consistency in the formats, it shouldn't take much to modify what I have already.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
michael-at-shaffer.net wrote: } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I use the term "metadata" loosely ... } } I'm looking into writing javascript for reading this SEM data (e.g., } magnification) that is embedded in the TIFF files in a non-standard way. I } expect the javascript to work within Photoshop, and I'd certainly be } interested if anyone has already done this and can provide an example } (although I am sure my SEM data is formatted differently). (BTW, I already } aware I can open these files with a text editor and simply extract the text } ... But that's not very elegant ...) } } The 2nd issue is what to do with the data once retrieved? I could simply } write it to a text file, but it would be better to write it back to the TIFF } in a standard way. For example, to put it all in the EXIF "comments" field } ... Or even better, to put it where it should be put ... Except I can find } no information as to Microscopists, as a group, asking that standard EXIF } fields be designated and standardized (e.g., the "microns per pixel" field). } } TIA :o) } } Cheerios, Michael Shaffer :o) } } SEM/MLA Research Coordinator } (709) 737-6799 (ofc) } (709) 737-6790 (lab) } (709) 737-6193 (FAX) } http://www.mun.ca/creait/maf/ } http://www.esd.mun.ca/epma/ } } Inco Innovation Centre } c/o Memorial University } 230 Elizabeth Avenue } P.O. Box 4200 } St. John's, NL A1C 5S7 } } } } } ==============================Original Headers============================== } 10, 19 -- From michael-at-shaffer.net Thu Mar 23 05:18:14 2006 } 10, 19 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) } 10, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NBIEjX025905 } 10, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 05:18:14 -0600 } 10, 19 -- Received: (qmail 6069 invoked from network); 23 Mar 2006 11:21:52 -0000 } 10, 19 -- Received: from unknown (HELO rarewolf1) (michael-at-shaffer.net-at-205.251.84.119) } 10, 19 -- by ws6-4.us4.outblaze.com with SMTP; 23 Mar 2006 11:21:52 -0000 } 10, 19 -- From: "michael shaffer" {michael-at-shaffer.net} } 10, 19 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} } 10, 19 -- Subject: SEM digital image "metadata" } 10, 19 -- Date: Thu, 23 Mar 2006 07:47:30 -0330 } 10, 19 -- Message-ID: {000d01c64e6b$612304f0$4f01a8c0-at-rarewolf1} } 10, 19 -- MIME-Version: 1.0 } 10, 19 -- Content-Type: text/plain; } 10, 19 -- charset="US-ASCII" } 10, 19 -- Content-Transfer-Encoding: 7bit } 10, 19 -- X-Mailer: Microsoft Office Outlook 11 } 10, 19 -- Thread-Index: AcZOa1+XpgOBO7+sRu65nUAmDJ856w== } 10, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 13, 19 -- From jehrman-at-mta.ca Thu Mar 23 07:27:46 2006 13, 19 -- Received: from mailserv.mta.ca (mailserv.mta.ca [138.73.1.1]) 13, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NDRkOX004970 13, 19 -- for {Microscopy-at-MSA.Microscopy.com} ; Thu, 23 Mar 2006 07:27:46 -0600 13, 19 -- Received: from host-22-245.mta.ca ([138.73.22.245]) 13, 19 -- by mailserv.mta.ca with esmtp (Exim 4.52) 13, 19 -- id 1FMPqm-00039t-MN; Thu, 23 Mar 2006 09:27:40 -0400 13, 19 -- Message-ID: {4422A245.8090801-at-mta.ca} 13, 19 -- Date: Thu, 23 Mar 2006 09:27:33 -0400 13, 19 -- From: "James M. Ehrman" {jehrman-at-mta.ca} 13, 19 -- Organization: Mount Allison University 13, 19 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 13, 19 -- MIME-Version: 1.0 13, 19 -- To: michael-at-shaffer.net, Microscopy Listserv {Microscopy-at-MSA.Microscopy.com} 13, 19 -- Subject: Re: [Microscopy] SEM digital image "metadata" 13, 19 -- References: {200603231118.k2NBItM9026890-at-ns.microscopy.com} 13, 19 -- In-Reply-To: {200603231118.k2NBItM9026890-at-ns.microscopy.com} 13, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Bleu Knight wrote: ================================================================== Does anyone have experience creating ultrathin sections of tissues that have bone or calcium crystals in them? If so, what kind of diamond knife are you using? I am reluctant to use my standard 45 degree diamond knife because I feel that it would dull very quickly. Thank you in advance for your response. ================================================================== So far as I know, all suppliers of diamond knives offer a version called a "materials science" diamond knife. However, some offer a product that has a higher knife angle (e.g. 55 instead of 45 deg.) but others, e.g. SPI offer the same angle (e.g. 45 deg) but with the caveat that the last of the "fine striations" have not been removed. We have never found these fine striations problematic since with the first pass of the knife over the sample, striations larger than these fine ones will be put into the knife edge anyhow. And since the final polishing step to take out the last of the fine striations is the most expensive, the cost of the SPI Supplies materials science knife is cheaper than that of a so-called "life science" knife.
We have found that the lower 45 deg angle results in sections easier to cut and with far fewer problems displaying "compression" effects.
You are correct in being reluctant to use your "standard" 45 deg knife because these kinds of samples will quite quickly put in striations that will render the knife useless for your other work.
You can find out more information about the SPI materials science diamond knife on URL http://www.2spi.com/catalog/knives/materials.shtml
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 15, 24 -- From cgarber-at-2spi.com Thu Mar 23 08:18:01 2006 15, 24 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 15, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NEI1Cs015158 15, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Mar 2006 08:18:01 -0600 15, 24 -- Received: from ibm1x23g2abfyg ([210.22.189.66]) 15, 24 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k2NEHlPI030752 15, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 23 Mar 2006 09:17:58 -0500 15, 24 -- X-IDV-FirstRcvd: [210.22.189.66] 15, 24 -- X-IDV-HELO: ibm1x23g2abfyg 15, 24 -- Message-ID: {012c01c64e84$9114d8f0$9f0aa8c0-at-ibm1x23g2abfyg} 15, 24 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 15, 24 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 15, 24 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 15, 24 -- Subject: Diamond knife for tissue with bone or calcium crystals 15, 24 -- Date: Thu, 23 Mar 2006 09:17:42 -0500 15, 24 -- MIME-Version: 1.0 15, 24 -- Content-Type: text/plain; 15, 24 -- charset="Windows-1252" 15, 24 -- X-Priority: 3 15, 24 -- X-MSMail-Priority: Normal 15, 24 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 15, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 15, 24 -- Content-Transfer-Encoding: 8bit 15, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2NEI1Cs015158 ==============================End of - Headers==============================
Thomas The answer is yes, tight junctions are a characteristic of vertebrate epithelia and as far as I know occur in all types whether keratinizing or not. Check your library for a copy of Porter & Bonneville's Fine Structure of Cells and Tissues for excellent EMs. Richard Harris Laboratory Supervisor, Imaging and Analysis Department of Biology, University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- X-from: phillipst-at-missouri.edu [mailto:phillipst-at-missouri.edu] Sent: Wednesday, March 22, 2006 7:47 PM To: rjharris-at-uwo.ca
Are there tight junctions (zonula occludens) in keratinized stratified squamous epithelia such as skin? Or do the lamellar granules (membrane coating granules) take care of all the sealing needed? As a corollary- are there tight junctions in non-keratinizing stratified epithelia such as esophagus? thanks for some basic histo I should know! tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
You may find that the magnification data is stored in your TIFF images in the XResolution and/or YResolution data which is a part of the standard TIFF structure. This data is stored as two long integers, the first representing the numerator, the second the denominator of a fractional number. There is another field called ResolutionUnit which gives the final result. The location of these fields within the TIFF file is found in directories in the file header. Noran used this method to store pixel size information in their Voyager/Vantage TIFF images. The full TIFF file specification can be found at http://partners.adobe.com/public/developer/en/tiff/TIFF6.pdf.
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==============================Original Headers============================== 6, 25 -- From djv23-at-cam.ac.uk Thu Mar 23 09:38:06 2006 6, 25 -- Received: from ppsw-1.csi.cam.ac.uk (ppsw-1.csi.cam.ac.uk [131.111.8.131]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NFc5vL002990 6, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 09:38:05 -0600 6, 25 -- X-Cam-SpamDetails: Not scanned 6, 25 -- X-Cam-AntiVirus: No virus found 6, 25 -- X-Cam-ScannerInfo: http://www.cam.ac.uk/cs/email/scanner/ 6, 25 -- Received: from focus.msm.cam.ac.uk ([131.111.100.73]:38831 helo=msm.cam.ac.uk) 6, 25 -- by ppsw-1.csi.cam.ac.uk (ppsw.cam.ac.uk [131.111.8.131]:25) 6, 25 -- with esmtp id 1FMRsm-0003cP-4K (Exim 4.54) 6, 25 -- (return-path {djv23-at-cam.ac.uk} ); Thu, 23 Mar 2006 15:37:52 +0000 6, 25 -- Received: from davids-pc.cam.ac.uk (sem-djv.msm.cam.ac.uk [131.111.100.208]) 6, 25 -- by msm.cam.ac.uk (8.13.6/8.13.6) with ESMTP id k2NFbnMR005393; 6, 25 -- Thu, 23 Mar 2006 15:37:52 GMT 6, 25 -- Message-Id: {6.2.1.2.0.20060323145258.02768bc0-at-focus.msm.cam.ac.uk} 6, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 6, 25 -- Date: Thu, 23 Mar 2006 15:40:38 +0000 6, 25 -- To: michael-at-shaffer.net 6, 25 -- From: David Vowles {djv23-at-cam.ac.uk} 6, 25 -- Subject: Re: [Microscopy] SEM digital image "metadata" 6, 25 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 6, 25 -- In-Reply-To: {200603231122.k2NBMF4o031271-at-ns.microscopy.com} 6, 25 -- References: {200603231122.k2NBMF4o031271-at-ns.microscopy.com} 6, 25 -- Mime-Version: 1.0 6, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Thanks to those with helpful comments about my question about whether there are tight junctions in either keratinized stratified squamous epithelia such as skin or non-keratinized stratified squamous epithelium such as esophagus. It is, as I expected, a controversial area. A tip led me to a couple of references from the Franke lab:
Langbein L. Grund C. Kuhn C. Praetzel S. Kartenbeck J. Brandner JM. Moll I. Franke WW. Tight junctions and compositionally related junctional structures in mammalian stratified epithelia and cell cultures derived therefrom. European Journal of Cell Biology. 81(8):419-35.
Schluter H. Wepf R. Moll I. Franke WW (2004) Sealing the live part of the skin: the integrated meshwork of desmosomes, tight junctions and curvilinear ridge structures in the cells of the uppermost granular layer of the human epidermis. European Journal of Cell Biology. 83(11-12):655-65.
I haven't gotten the full references yet but the abstracts state they "found an unexpected diversity of TJ-related structures" some of which show "colocalization with the most restricted transmembrane TJ marker protein, occludin,..." They report "TJ-related junctions are abundant..." is some stratified epithelia and that "most of them we have noticed, in addition, junctional regions showing relatively broad, ribbon-like membrane contacts which in cross-section often appear pentalaminar, with an electron-dense middle lamella ("lamellated TJs", coniunctiones laminosae)." Note that the abstract call these "TJ related junctions" and (since I haven't yet read the paper) implies to me that they are not necessarily classical TJ since they also note some of these are "characterized by a 10-30-nm dense lamina interposed between the two membranes" which is not something you would expect to see in a TJ. Thanks again for those who gave me tips to these and other references. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I am also interested to learn what methods are used in common brands of SEM to encode the scale or magnification information. We need this for our calibration and measurement software.
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
==============================Original Headers============================== 3, 21 -- From donc-at-asmicro.com Thu Mar 23 11:04:34 2006 3, 21 -- Received: from smtp109.sbc.mail.re2.yahoo.com (smtp109.sbc.mail.re2.yahoo.com [68.142.229.96]) 3, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NH4XFi023944 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:04:33 -0600 3, 21 -- Received: (qmail 98485 invoked from network); 23 Mar 2006 17:04:31 -0000 3, 21 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.251.105.88 with login) 3, 21 -- by smtp109.sbc.mail.re2.yahoo.com with SMTP; 23 Mar 2006 17:04:30 -0000 3, 21 -- Message-ID: {015601c64e9b$63af7b60$c901a8c0-at-ASM11} 3, 21 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- To: "Microscopy List" {microscopy-at-microscopy.com} 3, 21 -- Subject: SEM digital image "metadata" 3, 21 -- Date: Thu, 23 Mar 2006 12:00:58 -0500 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: text/plain; 3, 21 -- charset="iso-8859-1" 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Priority: 3 3, 21 -- X-MSMail-Priority: Normal 3, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 3, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
} You may find that the magnification data is stored in your } TIFF images in the XResolution and/or YResolution data which } is a part of the standard TIFF structure.
Thanx for your response David! For clarification, when you say "standard TIFF structure", are you claiming that if Photoshop opens the TIFF, and if you save it as a different file, the information is still there? The TIFF definition allows for many variations. Whether TIFF reads & writes recognize these variations is another question.
Inco Innovation Centre c/o Memorial University 230 Elizabeth Avenue P.O. Box 4200 St. John's, NL A1C 5S7
} } ------------------------------------------------------------- } ---------- } } ----- The Microscopy ListServer -- CoSponsor: The } Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ---------- } } ----- } } } } I use the term "metadata" loosely ... } } } } I'm looking into writing javascript for reading this SEM data (e.g., } } magnification) that is embedded in the TIFF files in a non-standard } } way. I expect the javascript to work within Photoshop, and I'd } } certainly be interested if anyone has already done this and } can provide } } an example (although I am sure my SEM data is formatted } differently). } } (BTW, I already aware I can open these files with a text editor and } } simply extract the text ... But that's not very elegant ...) } } } } The 2nd issue is what to do with the data once retrieved? I could } } simply write it to a text file, but it would be better to } write it back } } to the TIFF in a standard way. For example, to put it all } in the EXIF } } "comments" field ... Or even better, to put it where it } should be put } } ... Except I can find no information as to Microscopists, as } a group, } } asking that standard EXIF fields be designated and } standardized (e.g., the "microns per pixel" field). } } } } TIA :o) } } } } Cheerios, Michael Shaffer :o) }
==============================Original Headers============================== 10, 21 -- From Michael-at-Shaffer.net Thu Mar 23 11:11:58 2006 10, 21 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) 10, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NHBucs001070 10, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:11:57 -0600 10, 21 -- Received: (qmail 11789 invoked from network); 23 Mar 2006 17:15:41 -0000 10, 21 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 10, 21 -- by ws6-4.us4.outblaze.com with SMTP; 23 Mar 2006 17:15:41 -0000 10, 21 -- From: "michael shaffer" {michael-at-Shaffer.net} 10, 21 -- To: "'David Vowles'" {djv23-at-cam.ac.uk} 10, 21 -- Cc: "'MSA Microscopy list'" {Microscopy-at-microscopy.com} 10, 21 -- Subject: RE: [Microscopy] SEM digital image "metadata" 10, 21 -- Date: Thu, 23 Mar 2006 13:41:53 -0330 10, 21 -- Message-ID: {001e01c64e9c$e3150c70$8d829986-at-roamingwolf} 10, 21 -- MIME-Version: 1.0 10, 21 -- Content-Type: text/plain; 10, 21 -- charset="us-ascii" 10, 21 -- Content-Transfer-Encoding: 7bit 10, 21 -- X-Mailer: Microsoft Office Outlook 11 10, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 10, 21 -- thread-index: AcZOkExcOc847R7uRlefsxlIg0+GAgAC9nuA 10, 21 -- In-Reply-To: {6.2.1.2.0.20060323145258.02768bc0-at-focus.msm.cam.ac.uk} ==============================End of - Headers==============================
Cerium Labs LLC., a wholly owned subsidiary of Spansion Inc. (NASDAQ: SPSN, former Flash Memory groups of AMD and Fujitsu,) has an immediate opening for a TEM analyst. Lab operates 2 DB-FIBs and 2 TEMs (JEM-2010 and CM300FEG+GIF) and a wide range of analytical equipment. Emphasis is being placed on experience: analytical capabilities, EDS, GIF-EELS, and documented hands-on operation of TEM in various modes including electron crystallography. Lab is providing services to a range of clients from semiconductor, alternative energy, and advanced-materials sectors.
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Interested candidates, please feel free to forward to me your resumes and publication histories.
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Jerzy
****************************************************** Jerzy Gazda, Ph.D. CeriumLaboratories L.L.C.
Supervising Engineer 5204 E. Ben White Blvd. - MS 512 Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-ceriumlabs.com ******************************************************
==============================Original Headers============================== 13, 31 -- From Jerzy.Gazda-at-ceriumlabs.com Thu Mar 23 11:34:06 2006 13, 31 -- Received: from amdext4.amd.com (amdext4.amd.com [163.181.251.6]) 13, 31 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2NHY5nv010885 13, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:34:06 -0600 13, 31 -- Received: from SAUSGW02.amd.com (sausgw02.amd.com [163.181.250.22]) 13, 31 -- by amdext4.amd.com (8.12.11/8.12.11/AMD) with ESMTP id k2NHY6jC015807 13, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:34:06 -0600 13, 31 -- Received: from 163.181.22.101 by SAUSGW01.amd.com with ESMTP (AMD SMTP 13, 31 -- Relay (Email Firewall v6.1.0)); Thu, 23 Mar 2006 11:33:54 -0600 13, 31 -- X-Server-Uuid: 8C3DB987-180B-4465-9446-45C15473FD3E 13, 31 -- Received: from sausexmb2.amd.com ([163.181.3.157]) by sausexbh1.amd.com 13, 31 -- with Microsoft SMTPSVC(6.0.3790.0); Thu, 23 Mar 2006 09:33:54 -0800 13, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 13, 31 -- Content-class: urn:content-classes:message 13, 31 -- MIME-Version: 1.0 13, 31 -- Subject: Career Opportunity (UTZ231) 13, 31 -- Date: Thu, 23 Mar 2006 11:33:52 -0600 13, 31 -- Message-ID: {4E9FB73E2965BD41888C731E554E248303FE7A05-at-SAUSEXMB2.amd.com} 13, 31 -- X-MS-Has-Attach: 13, 31 -- X-MS-TNEF-Correlator: 13, 31 -- Thread-Topic: Career Opportunity (UTZ231) 13, 31 -- Thread-Index: AcZOn/Qpsoao9sTUScSb8rBuPdvLBQ== 13, 31 -- From: "Gazda, Jerzy" {Jerzy.Gazda-at-ceriumlabs.com} 13, 31 -- To: Microscopy-at-microscopy.com 13, 31 -- X-OriginalArrivalTime: 23 Mar 2006 17:33:54.0463 (UTC) 13, 31 -- FILETIME=[F50C52F0:01C64E9F] 13, 31 -- X-WSS-ID: 683C03880SS101799-01-01 13, 31 -- Content-Type: text/plain; 13, 31 -- charset=us-ascii 13, 31 -- Content-Transfer-Encoding: 8bit 13, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2NHY5nv010885 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cprrrw-at-msn.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 23, 2006 at 09:49:39 ---------------------------------------------------------------------------
Email: cprrrw-at-msn.com Name: Patricia VanLuven
Organization: Home school
Education: K-8 Grade Grammar School
Location: Laingsburg, Michigan, USA
Question: Can you recommend a good protozoa identification book for an upper elementary/middle school student. Some are quite expensive and only available on-line (without preview), so I am looking for a recommendation before I make a selection. Thank you very much. Patty VanLuven
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Rainis, K.G. and Russell, B.J. 1996 Guide to Microlife 287pp, 5.5x8.5", paperback, $40.00. ISBN 0-531-11266-7 Franklin Watts, Danbury, CT. The price of this book is both unfortunate and understandable. Unfortunate, because it should be in the library of every class that studies the microlife of our environment; understandable, because almost every page has one or more excellent color light micrographs. It's a comprehensive field guide to the microworld. The authors make the statement that the 115 microorganisms described comprise 75-90% of those that may be encountered in the "wild". The habitats described are diverse: the home, soils, plants and debris, and four aquatic environments, with detailed advice on collecting methods for each. Described organisms are equally diverse, ranging from monerans to millimeter-sized arthropods. Species descriptions include ecological information, advice on collection and culture, and frequent suggestions for further investigation. Middle school - adult. RECOMMENDED
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 4, 18 -- From schooley-at-mcn.org Thu Mar 23 18:34:46 2006 4, 18 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2O0YkQU004675 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 18:34:46 -0600 4, 18 -- Received: from [66.42.18.121] (helo=[10.0.1.35]) 4, 18 -- by dns4.mcn.org with esmtpa (Exim 4.60) 4, 18 -- (envelope-from {schooley-at-mcn.org} ) 4, 18 -- id IWLW9X-000FIU-2S; Thu, 23 Mar 2006 16:34:46 -0800 4, 18 -- Mime-Version: 1.0 4, 18 -- Message-Id: {a06200701c048ed65586d-at-[10.0.1.35]} 4, 18 -- In-Reply-To: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- References: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- Date: Thu, 23 Mar 2006 16:32:23 -0800 4, 18 -- To: cprrrw-at-msn.com 4, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 4, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: protozoa identification book 4, 18 -- Cc: microscopy-at-microscopy.com 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
There is no "standard" way for the EM manufacturers to encode magnification or calibration in the image or image metadata. Some manufacturers store the information there with other tags, usually private, used to store the scale in various forms, other vendors omit this entirely and provide an additional text file with the information. Also, using "standard" TIF tags can be hazardous. For example, some text processing programs like Word use the x-calibration value (or y-calibration) for calculating the size of the image on paper. If you put in the real calibration there, you might end up wit Word trying to print the image at the real size, and you end up with a dot!
In order to get this information, you need to contact the manufacturer and ask them for the format that they are using. This may or may not be information that they can give you. If you have several instruments, you probably need to do this for each one.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- X-from: donc-at-asmicro.com [mailto:donc-at-asmicro.com] Sent: Thursday, March 23, 2006 10:06 AM To: Mike Bode
I am also interested to learn what methods are used in common brands of SEM to encode the scale or magnification information. We need this for our calibration and measurement software.
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
==============================Original Headers============================== 3, 21 -- From donc-at-asmicro.com Thu Mar 23 11:04:34 2006 3, 21 -- Received: from smtp109.sbc.mail.re2.yahoo.com (smtp109.sbc.mail.re2.yahoo.com [68.142.229.96]) 3, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2NH4XFi023944 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 11:04:33 -0600 3, 21 -- Received: (qmail 98485 invoked from network); 23 Mar 2006 17:04:31 -0000 3, 21 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.251.105.88 with login) 3, 21 -- by smtp109.sbc.mail.re2.yahoo.com with SMTP; 23 Mar 2006 17:04:30 -0000 3, 21 -- Message-ID: {015601c64e9b$63af7b60$c901a8c0-at-ASM11} 3, 21 -- Reply-To: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 3, 21 -- To: "Microscopy List" {microscopy-at-microscopy.com} 3, 21 -- Subject: SEM digital image "metadata" 3, 21 -- Date: Thu, 23 Mar 2006 12:00:58 -0500 3, 21 -- MIME-Version: 1.0 3, 21 -- Content-Type: text/plain; 3, 21 -- charset="iso-8859-1" 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Priority: 3 3, 21 -- X-MSMail-Priority: Normal 3, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 3, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
==============================Original Headers============================== 13, 23 -- From Mike.Bode-at-soft-imaging.net Thu Mar 23 18:37:13 2006 13, 23 -- Received: from mail.soft-imaging.de (mail.soft-imaging.de [62.180.61.131]) 13, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2O0bCMh007749 13, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 18:37:12 -0600 13, 23 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 13, 23 -- by mail.soft-imaging.de (8.11.6/8.11.6/SuSE Linux 0.5) with ESMTP id k2O0bBD21654 13, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 01:37:11 +0100 13, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 23 -- Content-class: urn:content-classes:message 13, 23 -- MIME-Version: 1.0 13, 23 -- Content-Type: text/plain; 13, 23 -- charset="us-ascii" 13, 23 -- Subject: RE: [Microscopy] SEM digital image "metadata" 13, 23 -- Date: Fri, 24 Mar 2006 01:34:25 +0100 13, 23 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC94124FC3-at-ms-s-gws.soft-imaging.net} 13, 23 -- X-MS-Has-Attach: 13, 23 -- X-MS-TNEF-Correlator: 13, 23 -- Thread-Topic: [Microscopy] SEM digital image "metadata" 13, 23 -- Thread-Index: AcZOnBWMu/lqqZU6SrOOFRqh3pNidAAPbyog 13, 23 -- From: "Mike Bode" {Mike.Bode-at-soft-imaging.net} 13, 23 -- To: {Microscopy-at-microscopy.com} 13, 23 -- Content-Transfer-Encoding: 8bit 13, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2O0bCMh007749 ==============================End of - Headers==============================
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 mmanikka-at-umich.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mmanikka-at-umich.edu Name: Mohan Manikkam
Organization: University of Michigan
Title-Subject: [Filtered] Kodak NTB emulsion for autoradiography of in situ hybridized sections
Question: I am looking for the protocol and the experience of researchers with the Kodak NTB emulsion. I have used the previous versions, NTB-2 and NTB-3 but like to know if anyone has good results with the newer NTB emulsion with in situ hybridized tissue sections. I would appreciate members' valuable opinion and comment. Thanks.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both Bplowman-at-pacific.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: Bplowman-at-pacific.edu Name: Barbara Plowman
Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry
Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
If you open these dysfunctional TIF files with Photoshop, it will gloriously delete all the abnormal header info.
Zeiss has their own format which is deleted when the image is converted from indexed color to grey scale.
gary g.
At 09:13 AM 3/23/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Thu Mar 23 20:10:41 2006 10, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k2O2AfcF011966 10, 20 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 20:10:41 -0600 10, 20 -- Received: (qmail 14873 invoked from network); 23 Mar 2006 18:10:02 -0800 10, 20 -- Received: by simscan 1.1.0 ppid: 14870, pid: 14871, t: 0.1955s 10, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 20 -- by qsmtp4 with SMTP; 23 Mar 2006 18:10:02 -0800 10, 20 -- Message-Id: {6.2.3.4.2.20060323180759.023c98c0-at-mail.calweb.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 10, 20 -- Date: Thu, 23 Mar 2006 18:10:43 -0800 10, 20 -- To: michael-at-Shaffer.net 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] RE: SEM digital image "metadata" 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200603231713.k2NHDOgT004287-at-ns.microscopy.com} 10, 20 -- References: {200603231713.k2NHDOgT004287-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
briefly, I can tell you I am using acetonitrile (AN) as the "intermedium" after EtOH dehydration and embedding (as a substitute for PO) for now more than 15 years (human material, diagnostic and research specimens) without major problems in tissue &/or resin quality (polymerisation, cutting properties, stainability, stability in TEM-beam), provided you are aware of some specific properties of Acetonitrile.
As to my knowledge (and this was the cause for using AN instead of PO) Acetonitrile is stated "non-carcinogenic", despite being considered a mutagenic and cell toxic substance (PO is classified as "carcinogenic").
AN to 100% is water-miscible (so -theoretically- one should be able to use it as a substitute for EtOH as the dehydration solvent, but I've never tested that),
at ambient room and working conditions (humidity should not be too high, ventilated area needed like fume cupboard) you should get similar results for your specimen preparations, especially animal or human tissues (--} this was not the fact when I tried using so called "rapid dehydration methods" like the "acidified 2,2-DMP"-technique).
Vapor pressure of PO (unmiscible with water) is very high (-at-20 degr.C 588 hPa, -at- 33 degr.C 980 hPa, compared to AN: -at- 20 degr.C only 97 hPa; boiling point for PO: 35 degr.C, AN: 81 degr.C), "Highly flammable, vapors noxious, toxic if inhalated, swallowed or when contaminating skin"; MWC(1989): 40 ml/m3 - 70 mg/m3; Fresh water toxicity: Class 2 (do not waste into canalisation), but you will find all necessary physical data in the MSDS's provided with the substance delivered (hopefully!) (;:-))
for example see: http://www.jtbaker.com/msds/englishhtml/a0518.htm
(this was the } first { result at google)....you certainly will be "SHOCKED" but IMO: most of the chemicals used in (T)EM preparation do have some health risks if we don't work with them properly........(compare for that the statement of car producers: "Do not connect your exhaust pipe with the passenger room: don't inhale exhaust vapor...it might be lethal!") . Due to this "big" difference in vapor pressure, not only the substance's odours are "pleasant" as compared with PO.
Also, drying out of specimens during transfer of tissue into infiltration steps and pure resin (especially smallest ones) is not an issue any more. USE and Disposal of used solvent according to federal, national laws (in Europe/EC e.g. as "organic, non halogenated waste").
So - IMO - the most important thing: Using AN, you should be aware of a slower evaporation of solvent out of the tissue during infiltration (especially if room temperature is low) - thus you should
i) use specimen rotator(s)
ii) placing } infiltration { cups perhaps below a lamp (e.g. 60 W, at a distance of ca. 15 cm, temperature near specimens should be about 20-25 degr.C, see also v) below)
iii) placing specimens for "infiltration"-steps in flat "receptacula" (instead of [glass-] vials with a narrow neck and height of about 3 cm). For that purpose I fabricated on my own "special" infiltration silicone-rubber mo(u)lds (diam. ca. 1 cm, depth ca. 0.6 cm) which IMO are optimal for unhindered evaporation of the solvent, leaving also the option to polymerize the (pure) resin-fractions used for the infiltration-steps without any problem
iv) testing optimal infiltration times for the tissue you are embedding (usually, also for the big specimen blocks [up to 5 x 4 x 1 mm] I am working with, at least 45 min each infiltration step is -due to my experience - sufficient)
v) I use the following procedure (standardized for the diagnostic specimens, use of a specimen/probe rotator): - dehydration: ascending EtOH-series (50, 70, 80, 90, 96, 96, 100, 100, 100% EtOH),
- intermedium: 3 x pure AN (5, 10, 15 min), in ca. 5 ml snap cap vials, cap always closed (always take care of a surrounding humidity not to high !)
AN:Resin = 1: 1 : 1 x 45 minutes (at least), but that step can be elongated without problems up to 20 hours (i.e. e.g. over night, caps closed!), - before transfer into pure resin (into the infiltration moulds mentioned above, lamp) the cap of the vials is removed for at least 20-30 minutes (specimen rotator, under a lamp, see above),
pure RESIN: 3 x for at least 45 min each (use of lamp), take care of a humidity not to high !
There have been some papers related to the use of AN as a dehydration agent (and as a "safer" alternative to PO), if I remember correctly, in the 80ies or 90ies....if I find those or any in my files, I should be glad to share those informations with you (will take perhaps some hours of searching). One I have found by goo?gling: (http://www.emsdiasum.com/microscopy/technical/techtips/techtips2.aspx ) scroll down to # 18. Acetonitrile is a Safer, Better Dehydrating Solvent for Transmission Electron Microscopy: Propylene Oxide and Ethanol are commonly used dehydrating solvents for processing tissues for electron microscopy. But both solvents, however, have some undesirable properties: they are highly flammable, volatile, toxic and potentially carcinogenic. Acetonitrile is a direct substitution for ethanol and propylene oxide and it is safer to use and requires shorter dehydration times. It is freely miscible with water, alcohol, acetone and epoxy resin and it does not interfere with epoxy polymerization. Harold H. Edwards, Yu-Yan Yeh, Betty I. Tarnowsky, and Gregory R. Schonbaum. (1992). Acetonitrile as a Substitute for Ethanol/Propylene Oxide in Tissue Processing for Transmission Electron Microscopy: Comparison of Fine Structure and Lipid Solubility in Mouse Liver, Kidney, and Intestine. Microsc. Res. and Technique Vol. 21, pg.39-50
Hope this helps for now,
best regards and to all Listers: a beautiful Friday....weekend is coming (:-))
Wolfgang Muss
OR Dr. Wolfgang Muss EM-Lab =with pride: 25 years in operation by 2nd of Feb.2006= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at ------------------------------------------------------------------------ --------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------- Forthcoming Meetings:
SECOND ANNOUNCEMENT, REGISTER NOW ONLINE at: http://www.scur.org.pl
33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl -------------------------------- 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic -------------------------------- 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both Bplowman-at-pacific.edu as well as the MIcroscopy Listserver --------------------------------------------------------------------------- Email: Bplowman-at-pacific.edu Name: Barbara Plowman Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
} There is no "standard" way for the EM manufacturers to encode } magnification or calibration in the image or image metadata. } Some manufacturers store the information there with other } tags, usually private, used to store the scale in various } forms, other vendors omit this entirely and provide an } additional text file with the information.
Which begs the question "Why is there no standard method?" There is certainly enough EXIF fields still available. It would seem all that is needed is the push and for us to come up with a minimal number of field designations to apphoach the EXIF (or IPTC) people with.
} Also, using "standard" TIF tags can be hazardous. For } example, some text processing programs like Word use the } x-calibration value (or } y-calibration) for calculating the size of the image on } paper. If you put in the real calibration there, you might } end up wit Word trying to print the image at the real size, } and you end up with a dot!
Yes ... It's difficult enough to know why the EM manufacturers do not put the correct resolution into the file such that it'll print at the correct size. However, this TIFF field is not what I speaking of.
} In order to get this information, you need to contact the } manufacturer and ask them for the format that they are using. } This may or may not be information that they can give you. If } you have several instruments, you probably need to do this } for each one.
Personally, I have no need to do it for any other than my own SEM. However, if I am successful with the code, I'll get back to the group with the example. I know that at least a couple of SEM manufacturers are similar, if not the same.
Hello Everyone, I just purchased a copy of "Guide to Microlife" (how could I resist not looking at pond water and knowing what I'm looking at!) from Buy.com for under $24 including shipping. As an aside, they offered my a $25.00 discount coupon for my next order. One might consider ordering one book, then with the coupon order more...
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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schooley-at-mcn.org To: frank.karl-at-degussa.com 03/23/2006 07:36 cc: PM Subject: [Microscopy] Re: AskAMicroscopist: protozoa identification book Please respond to schooley
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} } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (cprrrw-at-msn.com) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Thursday, March 23, 2006 at 09:49:39 } ---------------------------------------------------------------------------
} } Email: cprrrw-at-msn.com } Name: Patricia VanLuven } } Organization: Home school } } Education: K-8 Grade Grammar School } } Location: Laingsburg, Michigan, USA } } Question: Can you recommend a good protozoa identification book for } an upper elementary/middle school student. Some are quite expensive } and only available on-line (without preview), so I am looking for a } recommendation before I make a selection. Thank you very much. } Patty VanLuven } } ---------------------------------------------------------------------------
Patricia - You may consider this too expensive, but it's what you need; maybe you can find a used copy. The description is taken from the MICRO bibliography (URL below).
Rainis, K.G. and Russell, B.J. 1996 Guide to Microlife 287pp, 5.5x8.5", paperback, $40.00. ISBN 0-531-11266-7 Franklin Watts, Danbury, CT. The price of this book is both unfortunate and understandable. Unfortunate, because it should be in the library of every class that studies the microlife of our environment; understandable, because almost every page has one or more excellent color light micrographs. It's a comprehensive field guide to the microworld. The authors make the statement that the 115 microorganisms described comprise 75-90% of those that may be encountered in the "wild". The habitats described are diverse: the home, soils, plants and debris, and four aquatic environments, with detailed advice on collecting methods for each. Described organisms are equally diverse, ranging from monerans to millimeter-sized arthropods. Species descriptions include ecological information, advice on collection and culture, and frequent suggestions for further investigation. Middle school - adult. RECOMMENDED
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 4, 18 -- From schooley-at-mcn.org Thu Mar 23 18:34:46 2006 4, 18 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 4, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2O0YkQU004675 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 23 Mar 2006 18:34:46 -0600 4, 18 -- Received: from [66.42.18.121] (helo=[10.0.1.35]) 4, 18 -- by dns4.mcn.org with esmtpa (Exim 4.60) 4, 18 -- (envelope-from {schooley-at-mcn.org} ) 4, 18 -- id IWLW9X-000FIU-2S; Thu, 23 Mar 2006 16:34:46 -0800 4, 18 -- Mime-Version: 1.0 4, 18 -- Message-Id: {a06200701c048ed65586d-at-[10.0.1.35]} 4, 18 -- In-Reply-To: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- References: {200603232334.k2NNYvOt003568-at-ns.microscopy.com} 4, 18 -- Date: Thu, 23 Mar 2006 16:32:23 -0800 4, 18 -- To: cprrrw-at-msn.com 4, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 4, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: protozoa identification book 4, 18 -- Cc: microscopy-at-microscopy.com 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 29, 18 -- From frank.karl-at-degussa.com Fri Mar 24 07:48:26 2006 29, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 29, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2ODmOCH020633 29, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Mar 2006 07:48:25 -0600 29, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 29, 18 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2ODmHpE021422; 29, 18 -- Fri, 24 Mar 2006 14:48:18 +0100 29, 18 -- In-Reply-To: {200603240036.k2O0ac4B007038-at-ns.microscopy.com} 29, 18 -- Subject: Re: AskAMicroscopist: protozoa identification book 29, 18 -- To: schooley-at-mcn.org, microscopy-at-msa.microscopy.com 29, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 29, 18 -- Message-ID: {OF46EF54FD.85491A20-ON8525713B.004B5C62-8525713B.004BD1C7-at-degussa.com} 29, 18 -- From: frank.karl-at-degussa.com 29, 18 -- Date: Fri, 24 Mar 2006 08:48:09 -0500 29, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 29, 18 -- 03/24/2006 07:48:20 AM 29, 18 -- MIME-Version: 1.0 29, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
The "Guide to Microlife" is $23 from amazon.com. A better, but more expensive (but without all the color photographs) is Theodore Jahn, et al. "How to Know the Protozoa". The classification is outdated, since its 1978, but it's still a useful guide. Should be available used from Advanced Book Exchange (abebooks.com), Powell's, and the like. But be careful! Many of the used copies are the 1948 or 1963 editions. Better still is Patterson's "Free-living Freshwater Protozoa: A Color Guide", but this is $60. Unfortunately, that's cheap anymore for books.
Phil
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-- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Fri Mar 24 08:15:56 2006 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2OEFt49030656 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 08:15:55 -0600 4, 22 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k2OEpR4v008018 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 09:53:36 -0500 4, 22 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 4, 22 -- Fri, 24 Mar 2006 09:08:27 -0500 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230901c049a9a79f16-at-[141.209.160.132]} 4, 22 -- In-Reply-To: {200603240040.k2O0exLx020415-at-ns.microscopy.com} 4, 22 -- References: {200603240040.k2O0exLx020415-at-ns.microscopy.com} 4, 22 -- Date: Fri, 24 Mar 2006 09:08:39 -0500 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: [Microscopy] Re: AskAMicroscopist: protozoa identification book 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 24 Mar 2006 14:08:27.0851 (UTC) FILETIME=[6C3A59B0:01C64F4C] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -4 () L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
The Minnesota Microscopy Society (MMS) will hold its annual Spring Symposium on April 21st. This year's topic is "Microcopy in Failure Analysis". The all day event will be held at the Minnesota Science Museum.
Topic will include:
Plastic Component Failure Analysis Failure Analysis in the 21st Century, Nano-Scale Materials Specimen Selection in Microscopy for Failure Analysis Use of the SEM in the Failure Analysis of Cardiac Pacing Leads Vendor Displays
Cost $75 per person for memember and $85 per person for non members. Lunch and coffee breaks provided.
Reservations MUST be made no later than Friday, April 14th. Register by e-mailing Bede Willenbring at Bede.Willenbring-at-hbfuller.com, or by phone at 651-236-5470. Include your name, company, phone number, and e-mail address.
Full details are available online at http://www.MNmicroscopy.org - just click on the "current newsletter" in your preferred format; html for browsing or pdf for printing out to show your colleagues.
Thank you
Minnesota Microscopy Society
Prof. Stuart McKernan IT Characterization Facility, University of Minnesota E-mail : stuartm-at-umn.edu 12 Shepherd Labs, Office: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 626-7594
==============================Original Headers============================== 13, 15 -- From stuartm-at-umn.edu Fri Mar 24 09:22:57 2006 13, 15 -- Received: from mtaout-w.tc.umn.edu (mtaout-w.tc.umn.edu [160.94.160.21]) 13, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2OFMvoG009757 13, 15 -- for {microscopy-at-microscopy.com} ; Fri, 24 Mar 2006 09:22:57 -0600 13, 15 -- Received: from [160.94.16.136] (cfsigma.charfac.umn.edu [160.94.16.136]) by mtaout-w.tc.umn.edu with ESMTP for microscopy-at-microscopy.com; Fri, 24 Mar 2006 09:22:49 -0600 (CST) 13, 15 -- X-Umn-Remote-Mta: [N] cfsigma.charfac.umn.edu [160.94.16.136] #+LO+TS+AU+HN 13, 15 -- Mime-Version: 1.0 (Apple Message framework v746.3) 13, 15 -- Content-Transfer-Encoding: 7bit 13, 15 -- Message-Id: {E6603809-4267-4B31-8C2F-132ED229FE51-at-umn.edu} 13, 15 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 13, 15 -- To: microscopy-at-microscopy.com 13, 15 -- From: stuart McKernan {stuartm-at-umn.edu} 13, 15 -- Subject: MMS Spring Symposium 13, 15 -- Date: Fri, 24 Mar 2006 09:22:47 -0600 13, 15 -- X-Mailer: Apple Mail (2.746.3) ==============================End of - Headers==============================
Sir, it is my duty, to distribute this your valued information to the MSA-listers. I am sorry if I "encouraged" somebody to use that stuff without cautions you are stating below. I always adhered to GLP procedures, in this special case by using } fume cupboard {. Also I found a hint on the toxic effects of ACN(Acetonitrile) when combined with water in the List Server Archives, July 1994 by Marcelle A Gillott, who stated: *** CAUTION *** acetonitrile combined with water releases hydrogen cyanide gas !!!
while it is touted as being considerably less toxic than PO users should be aware of the above reaction if it is being used as a dehydrant.
I shall use ACN from now on more carefully than ever.
Thank you for your highly valued comment !
Best regards Wolfgang Muss
PS: another issue for people working with fixatives - similar to this "hidden ACN problem" - would be formaldehyde (as used in tissue fixation procedures) and traces of hydrochloric acid ==} producing highly toxic and cancerogenic Bis-Chloromethylether (bis-CME)....
First of all, I do not disagree with any of your statements on the safety comparison between acetonitrile and propylene oxide.
However I do feel that you have subsequently understated the hazards of acetonitrile.
Acetonitrile (ACN) and Acrylonitrile (AN) are both significantly hazardous chemicals in that they break down in the body and generate hydrogen cyanide, HCN. Both AN and ACN are readily absorbed through skin and tissues. Cyanide poisoning has many stages, none of which are good! Please be careful with these very useful, but hazardous solvents.
As you have previously mentioned, Use of these solvents in your lab should be done carefully, and firstly with SOPs and engineering controls well established and implemented. Also with resulting PPE verified, and employed. Depending on the quantities and concentrations, you may want to consider keeping one of several HCN antidotes on hand, or make your emergency response system aware of your use of these chemicals so that they can have the antidote on hand. Many fire departments and paramedics units carry HCN antidotes or have a special first-aid treatment for victims exposed to chemicals in this family.
Please communicate this info to your colleagues working with these chemicals, and by all means consult the MSDS for every chemical that you plan to use.
Brad Huggins BP Chemicals Naperville, IL
Sometimes, being careful
Some more info below:
Response of Humans to HCN in Air 270 ppm Immediately fatal 181 ppm Fatal after 10 minutes 135 ppm Fatal after 30 minutes 110-135 ppm Fatal 30-60+ minutes 45-55 ppm Tolerated for 30-60 min
Early Physical Findings with Contaminated Victim: Special caution for Head, Ear, Eye, Nose and Mouth/Throat Bright red retinal veins and arteries Smell of bitter almonds on the breath
Cyanide antidotes if diagnosis is certain: Sodium nitrite intravenously Sodium thiosulfite intravenously Kelocyanor available in UK and France Amyl nitrite ampoules: temporizing therapy until IV access is obtained
Subchronic Toxicity Sporadic vapor exposure for 6 years: Loss of appetite, nervousness, vertigo, headache, nausea, vomiting Goldsmith apprentice: Headache, listlessness, numbness, partial paralysis of left arm and leg, partial loss of vision left eye, and EKG abnormalities
-----Original Message----- X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at] Sent: Friday, March 24, 2006 3:19 AM To: Huggins, Bradley J
Good morning, dear Barbara,
briefly, I can tell you I am using acetonitrile (AN) as the "intermedium" after EtOH dehydration and embedding (as a substitute for PO) for now more than 15 years (human material, diagnostic and research specimens) without major problems in tissue &/or resin quality (polymerisation, cutting properties, stainability, stability in TEM-beam), provided you are aware of some specific properties of Acetonitrile.
As to my knowledge (and this was the cause for using AN instead of PO) Acetonitrile is stated "non-carcinogenic", despite being considered a mutagenic and cell toxic substance (PO is classified as "carcinogenic").
AN to 100% is water-miscible (so -theoretically- one should be able to use it as a substitute for EtOH as the dehydration solvent, but I've never tested that),
at ambient room and working conditions (humidity should not be too high, ventilated area needed like fume cupboard) you should get similar results for your specimen preparations, especially animal or human tissues (--} this was not the fact when I tried using so called "rapid dehydration methods" like the "acidified 2,2-DMP"-technique).
Vapor pressure of PO (unmiscible with water) is very high (-at-20 degr.C 588 hPa, -at- 33 degr.C 980 hPa, compared to AN: -at- 20 degr.C only 97 hPa; boiling point for PO: 35 degr.C, AN: 81 degr.C), "Highly flammable, vapors noxious, toxic if inhalated, swallowed or when contaminating skin"; MWC(1989): 40 ml/m3 - 70 mg/m3; Fresh water toxicity: Class 2 (do not waste into canalisation), but you will find all necessary physical data in the MSDS's provided with the substance delivered (hopefully!) (;:-))
for example see: http://www.jtbaker.com/msds/englishhtml/a0518.htm
(this was the } first { result at google)....you certainly will be "SHOCKED" but IMO: most of the chemicals used in (T)EM preparation do have some health risks if we don't work with them properly........(compare for that the statement of car producers: "Do not connect your exhaust pipe with the passenger room: don't inhale exhaust vapor...it might be lethal!") . Due to this "big" difference in vapor pressure, not only the substance's odours are "pleasant" as compared with PO.
Also, drying out of specimens during transfer of tissue into infiltration steps and pure resin (especially smallest ones) is not an issue any more. USE and Disposal of used solvent according to federal, national laws (in Europe/EC e.g. as "organic, non halogenated waste").
So - IMO - the most important thing: Using AN, you should be aware of a slower evaporation of solvent out of the tissue during infiltration (especially if room temperature is low) - thus you should
i) use specimen rotator(s)
ii) placing } infiltration { cups perhaps below a lamp (e.g. 60 W, at a distance of ca. 15 cm, temperature near specimens should be about 20-25 degr.C, see also v) below)
iii) placing specimens for "infiltration"-steps in flat "receptacula" (instead of [glass-] vials with a narrow neck and height of about 3 cm).
For that purpose I fabricated on my own "special" infiltration silicone-rubber mo(u)lds (diam. ca. 1 cm, depth ca. 0.6 cm) which IMO are optimal for unhindered evaporation of the solvent, leaving also the option to polymerize the (pure) resin-fractions used for the infiltration-steps without any problem
iv) testing optimal infiltration times for the tissue you are embedding (usually, also for the big specimen blocks [up to 5 x 4 x 1 mm] I am working with, at least 45 min each infiltration step is -due to my experience - sufficient)
v) I use the following procedure (standardized for the diagnostic specimens, use of a specimen/probe rotator): - dehydration: ascending EtOH-series (50, 70, 80, 90, 96, 96, 100, 100, 100% EtOH),
- intermedium: 3 x pure AN (5, 10, 15 min), in ca. 5 ml snap cap vials, cap always closed (always take care of a surrounding humidity not to high !)
AN:Resin = 1: 1 : 1 x 45 minutes (at least), but that step can be elongated without problems up to 20 hours (i.e. e.g. over night, caps closed!), - before transfer into pure resin (into the infiltration moulds mentioned above, lamp) the cap of the vials is removed for at least 20-30 minutes (specimen rotator, under a lamp, see above),
pure RESIN: 3 x for at least 45 min each (use of lamp), take care of a humidity not to high !
There have been some papers related to the use of AN as a dehydration agent (and as a "safer" alternative to PO), if I remember correctly, in the 80ies or 90ies....if I find those or any in my files, I should be glad to share those informations with you (will take perhaps some hours of searching). One I have found by goo?gling: (http://www.emsdiasum.com/microscopy/technical/techtips/techtips2.aspx ) scroll down to # 18. Acetonitrile is a Safer, Better Dehydrating Solvent for Transmission Electron Microscopy: Propylene Oxide and Ethanol are commonly used dehydrating solvents for processing tissues for electron microscopy. But both solvents, however, have some undesirable properties: they are highly flammable, volatile, toxic and potentially carcinogenic. Acetonitrile is a direct substitution for ethanol and propylene oxide and it is safer to use and requires shorter dehydration times. It is freely miscible with water, alcohol, acetone and epoxy resin and it does not interfere with epoxy polymerization. Harold H. Edwards, Yu-Yan Yeh, Betty I. Tarnowsky, and Gregory R. Schonbaum. (1992). Acetonitrile as a Substitute for Ethanol/Propylene Oxide in Tissue Processing for Transmission Electron Microscopy: Comparison of Fine Structure and Lipid Solubility in Mouse Liver, Kidney, and Intestine. Microsc. Res. and Technique Vol. 21, pg.39-50
Hope this helps for now,
best regards and to all Listers: a beautiful Friday....weekend is coming (:-))
Wolfgang Muss
OR Dr. Wolfgang Muss EM-Lab =with pride: 25 years in operation by 2nd of Feb.2006= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at ------------------------------------------------------------------------ --------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------ - Forthcoming Meetings:
SECOND ANNOUNCEMENT, REGISTER NOW ONLINE at: http://www.scur.org.pl
33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl -------------------------------- 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic -------------------------------- 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
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---- This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both Bplowman-at-pacific.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ --- Email: Bplowman-at-pacific.edu Name: Barbara Plowman Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or
acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
Just about all procedures using chemicals commonly found in electron microscopy should be performed in fume hoods. All scientists and support staff should read the appropriate Material Safety Data Sheets (MSDS) for each chemical used (many online sites). Save copies of these for future reference/emergencies/etc. and update every several years. A 10-15 year old MSDS may be useless.
Of course there will be exceptions but use "common sense" [:)]. Expect your safety guy/gal to lack this quality.
Even old dogs learn new tricks!
X-from the New Orleans Diaspora,
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, MSA/SRRC SWSRU P.O. Box 350 Stoneville, MS 38776
} } } {W.Muss-at-salk.at} 03/24/06 11:41AM } } } ---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Sir, it is my duty, to distribute this your valued information to the MSA-listers. I am sorry if I "encouraged" somebody to use that stuff without cautions you are stating below. I always adhered to GLP procedures, in this special case by using } fume cupboard {. Also I found a hint on the toxic effects of ACN(Acetonitrile) when combined with water in the List Server Archives, July 1994 by Marcelle A Gillott, who stated: *** CAUTION *** acetonitrile combined with water releases hydrogen cyanide gas !!!
while it is touted as being considerably less toxic than PO users should be aware of the above reaction if it is being used as a dehydrant.
I shall use ACN from now on more carefully than ever.
Thank you for your highly valued comment !
Best regards Wolfgang Muss
PS: another issue for people working with fixatives - similar to this "hidden ACN problem" - would be formaldehyde (as used in tissue fixation procedures) and traces of hydrochloric acid ==} producing highly toxic and cancerogenic Bis-Chloromethylether (bis-CME)....
First of all, I do not disagree with any of your statements on the safety comparison between acetonitrile and propylene oxide.
However I do feel that you have subsequently understated the hazards of acetonitrile.
Acetonitrile (ACN) and Acrylonitrile (AN) are both significantly hazardous chemicals in that they break down in the body and generate hydrogen cyanide, HCN. Both AN and ACN are readily absorbed through skin and tissues. Cyanide poisoning has many stages, none of which are good! Please be careful with these very useful, but hazardous solvents.
As you have previously mentioned, Use of these solvents in your lab should be done carefully, and firstly with SOPs and engineering controls well established and implemented. Also with resulting PPE verified, and employed. Depending on the quantities and concentrations, you may want to consider keeping one of several HCN antidotes on hand, or make your emergency response system aware of your use of these chemicals so that they can have the antidote on hand. Many fire departments and paramedics units carry HCN antidotes or have a special first-aid treatment for victims exposed to chemicals in this family.
Please communicate this info to your colleagues working with these chemicals, and by all means consult the MSDS for every chemical that you plan to use.
Brad Huggins BP Chemicals Naperville, IL
Sometimes, being careful
Some more info below:
Response of Humans to HCN in Air 270 ppm Immediately fatal 181 ppm Fatal after 10 minutes 135 ppm Fatal after 30 minutes 110-135 ppm Fatal 30-60+ minutes 45-55 ppm Tolerated for 30-60 min
Early Physical Findings with Contaminated Victim: Special caution for Head, Ear, Eye, Nose and Mouth/Throat Bright red retinal veins and arteries Smell of bitter almonds on the breath
Cyanide antidotes if diagnosis is certain: Sodium nitrite intravenously Sodium thiosulfite intravenously Kelocyanor available in UK and France Amyl nitrite ampoules: temporizing therapy until IV access is obtained
Subchronic Toxicity Sporadic vapor exposure for 6 years: Loss of appetite, nervousness, vertigo, headache, nausea, vomiting Goldsmith apprentice: Headache, listlessness, numbness, partial paralysis of left arm and leg, partial loss of vision left eye, and EKG abnormalities
-----Original Message----- X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at] Sent: Friday, March 24, 2006 3:19 AM To: Huggins, Bradley J
Good morning, dear Barbara,
briefly, I can tell you I am using acetonitrile (AN) as the "intermedium" after EtOH dehydration and embedding (as a substitute for PO) for now more than 15 years (human material, diagnostic and research specimens) without major problems in tissue &/or resin quality (polymerisation, cutting properties, stainability, stability in TEM-beam), provided you are aware of some specific properties of Acetonitrile.
As to my knowledge (and this was the cause for using AN instead of PO) Acetonitrile is stated "non-carcinogenic", despite being considered a mutagenic and cell toxic substance (PO is classified as "carcinogenic").
AN to 100% is water-miscible (so -theoretically- one should be able to use it as a substitute for EtOH as the dehydration solvent, but I've never tested that),
at ambient room and working conditions (humidity should not be too high, ventilated area needed like fume cupboard) you should get similar results for your specimen preparations, especially animal or human tissues (--} this was not the fact when I tried using so called "rapid dehydration methods" like the "acidified 2,2-DMP"-technique).
Vapor pressure of PO (unmiscible with water) is very high (-at-20 degr.C 588 hPa, -at- 33 degr.C 980 hPa, compared to AN: -at- 20 degr.C only 97 hPa; boiling point for PO: 35 degr.C, AN: 81 degr.C), "Highly flammable, vapors noxious, toxic if inhalated, swallowed or when contaminating skin"; MWC(1989): 40 ml/m3 - 70 mg/m3; Fresh water toxicity: Class 2 (do not waste into canalisation), but you will find all necessary physical data in the MSDS's provided with the substance delivered (hopefully!) (;:-))
for example see: http://www.jtbaker.com/msds/englishhtml/a0518.htm
(this was the } first { result at google)....you certainly will be "SHOCKED" but IMO: most of the chemicals used in (T)EM preparation do have some health risks if we don't work with them properly........(compare for that the statement of car producers: "Do not connect your exhaust pipe with the passenger room: don't inhale exhaust vapor...it might be lethal!") . Due to this "big" difference in vapor pressure, not only the substance's odours are "pleasant" as compared with PO.
Also, drying out of specimens during transfer of tissue into infiltration steps and pure resin (especially smallest ones) is not an issue any more. USE and Disposal of used solvent according to federal, national laws (in Europe/EC e.g. as "organic, non halogenated waste").
So - IMO - the most important thing: Using AN, you should be aware of a slower evaporation of solvent out of the tissue during infiltration (especially if room temperature is low) - thus you should
i) use specimen rotator(s)
ii) placing } infiltration { cups perhaps below a lamp (e.g. 60 W, at a distance of ca. 15 cm, temperature near specimens should be about 20-25 degr.C, see also v) below)
iii) placing specimens for "infiltration"-steps in flat "receptacula" (instead of [glass-] vials with a narrow neck and height of about 3 cm).
For that purpose I fabricated on my own "special" infiltration silicone-rubber mo(u)lds (diam. ca. 1 cm, depth ca. 0.6 cm) which IMO are optimal for unhindered evaporation of the solvent, leaving also the option to polymerize the (pure) resin-fractions used for the infiltration-steps without any problem
iv) testing optimal infiltration times for the tissue you are embedding (usually, also for the big specimen blocks [up to 5 x 4 x 1 mm] I am working with, at least 45 min each infiltration step is -due to my experience - sufficient)
v) I use the following procedure (standardized for the diagnostic specimens, use of a specimen/probe rotator): - dehydration: ascending EtOH-series (50, 70, 80, 90, 96, 96, 100, 100, 100% EtOH),
- intermedium: 3 x pure AN (5, 10, 15 min), in ca. 5 ml snap cap vials, cap always closed (always take care of a surrounding humidity not to high !)
AN:Resin = 1: 1 : 1 x 45 minutes (at least), but that step can be elongated without problems up to 20 hours (i.e. e.g. over night, caps closed!), - before transfer into pure resin (into the infiltration moulds mentioned above, lamp) the cap of the vials is removed for at least 20-30 minutes (specimen rotator, under a lamp, see above),
pure RESIN: 3 x for at least 45 min each (use of lamp), take care of a humidity not to high !
There have been some papers related to the use of AN as a dehydration agent (and as a "safer" alternative to PO), if I remember correctly, in the 80ies or 90ies....if I find those or any in my files, I should be glad to share those informations with you (will take perhaps some hours of searching). One I have found by goo?gling: (http://www.emsdiasum.com/microscopy/technical/techtips/techtips2.aspx ) scroll down to # 18. Acetonitrile is a Safer, Better Dehydrating Solvent for Transmission Electron Microscopy: Propylene Oxide and Ethanol are commonly used dehydrating solvents for processing tissues for electron microscopy. But both solvents, however, have some undesirable properties: they are highly flammable, volatile, toxic and potentially carcinogenic. Acetonitrile is a direct substitution for ethanol and propylene oxide and it is safer to use and requires shorter dehydration times. It is freely miscible with water, alcohol, acetone and epoxy resin and it does not interfere with epoxy polymerization. Harold H. Edwards, Yu-Yan Yeh, Betty I. Tarnowsky, and Gregory R. Schonbaum. (1992). Acetonitrile as a Substitute for Ethanol/Propylene Oxide in Tissue Processing for Transmission Electron Microscopy: Comparison of Fine Structure and Lipid Solubility in Mouse Liver, Kidney, and Intestine. Microsc. Res. and Technique Vol. 21, pg.39-50
Hope this helps for now,
best regards and to all Listers: a beautiful Friday....weekend is coming (:-))
Wolfgang Muss
OR Dr. Wolfgang Muss EM-Lab =with pride: 25 years in operation by 2nd of Feb.2006= Institute of Pathology, SALK (Salzburger Landeskliniken gemeinnuetzige GesmbH) Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/EUROPE
and/or/alternatively
Paracelsus Medical Private University (PMU) Institute of Pathology Electron Microscopy Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone work: +43+662+4482+4720 Mobile phone work:+43+662+4482-57704 Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss") E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456 E-Mail private: wij.Muss-at-aon.at ------------------------------------------------------------------------ --------------------------------Information on behalf of Society for Cutaneous Ultrastructure Research (SCUR) PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org { ------------------------------------------------------------------------ - Forthcoming Meetings:
SECOND ANNOUNCEMENT, REGISTER NOW ONLINE at: http://www.scur.org.pl
33rd Annual Meeting of the SCUR, 8th-10th June, 2006, WARSZAW, Poland WEBSITE, containing all FORMS: http://www.scur.org.pl Additional informations: send an E-Mail kwoznia-at-amwaw.edu.pl -------------------------------- 34th Annual Meeting of the SCUR, 11th-12th May, 2007, PRAGUE Czech Republic -------------------------------- 35th Annual Meeting of the SCUR, 11th -12th May, 2008, NARA or KYOTO, Japan Joint Meeting with the JSUCB, the Japanese Society for Ultrastructural Cutaneous Biology
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---- This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both Bplowman-at-pacific.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ --- Email: Bplowman-at-pacific.edu Name: Barbara Plowman Organization: Univ of the Pacific, Arthur A. Dugoni School of Dentistry Title-Subject: [Filtered] acetonitrile
Question: I need information about acetonitrile as a substitute for propylene oxide. Is it carcinogenic? Is it used for dehydration like alcohol? Is it safer than P.O.? Am I better off with propylene oxide or
acetonitrile? (I am using this for salivary glands in rats) Thanks in advance. Barbara Plowman
Univ. of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster Rm 642 San Francisco, CA 94115
I got a question I hope the collective wisdom can help answer. I have TiO2 in an organic matrix. PLM shows the expected optical properties, but I would like to run a microchemical test to confirm the presence of Ti. I seem to remember a bead test to fuse the TiO2 into something water soluble followed by a microchemical with ? quinoline and ammonium thiocyanate? Squaric acid?
Any suggestion to get my TiO2 into solution would be welcome!
Thanks!! Frank
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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==============================Original Headers============================== 9, 17 -- From frank.karl-at-degussa.com Fri Mar 24 13:36:48 2006 9, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 9, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k2OJalsF010151 9, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Mar 2006 13:36:47 -0600 9, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 9, 17 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2OJaiue027149 9, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 24 Mar 2006 20:36:45 +0100 9, 17 -- Subject: Question Microchemical test for Ti 9, 17 -- To: microscopy-at-msa.microscopy.com 9, 17 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 9, 17 -- Message-ID: {OF6F9B1405.4F061B37-ON8525713B.006A27A0-8525713B.006BB8D7-at-degussa.com} 9, 17 -- From: frank.karl-at-degussa.com 9, 17 -- Date: Fri, 24 Mar 2006 14:36:36 -0500 9, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 9, 17 -- 03/24/2006 01:36:46 PM 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (streiker-at-sbcglobal.net) from on Saturday, March 25, 2006 at 20:13:39 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both streiker-at-sbcglobal.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: streiker-at-sbcglobal.net Name: Scott Streiker
Organization: University of Dayton
Education: Graduate College
Location: Dayton, Ohio, USA
Title: Atomic lattice with TEM
Question: What is the protocol for using a TEM to view atomic lattice/planes of carbon nano tubes at direct magnification over 200K at accel voltage of 100kV or greater?
which seems to describe the use of potassium thiocarbonate.
I don't know if anyone uses ring oven techniques any more, they were widely developed and promoted by Phil West and his wife in Baton Rouge in the 1960s and 1970s and at the time seemed to me very interesting. Then I changed jobs.
cheers
rtch
On 24 Mar 2006 at 13:38, frank.karl-at-degussa.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I got a question I hope the collective wisdom can help answer. I have TiO2 } in an organic matrix. PLM shows the expected optical properties, but I } would like to run a microchemical test to confirm the presence of Ti. I } seem to remember a bead test to fuse the TiO2 into something water soluble } followed by a microchemical with ? quinoline and ammonium thiocyanate? } Squaric acid? } } Any suggestion to get my TiO2 into solution would be welcome! } } Thanks!! Frank } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 13, 27 -- From r.sims-at-auckland.ac.nz Sun Mar 26 17:08:49 2006 13, 27 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) 13, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2QN8mro017444 13, 27 -- for {microscopy-at-msa.microscopy.com} ; Sun, 26 Mar 2006 17:08:49 -0600 13, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 13, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 57B9634D50; 13, 27 -- Mon, 27 Mar 2006 11:08:47 +1200 (NZST) 13, 27 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) 13, 27 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 13, 27 -- with ESMTP id 21609-16; Mon, 27 Mar 2006 11:08:47 +1200 (NZST) 13, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 13, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 0810934E65; 13, 27 -- Mon, 27 Mar 2006 11:08:45 +1200 (NZST) 13, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 13, 27 -- To: frank.karl-at-degussa.com 13, 27 -- Date: Mon, 27 Mar 2006 11:12:15 +1200 13, 27 -- MIME-Version: 1.0 13, 27 -- Subject: Re: [Microscopy] Question Microchemical test for Ti 13, 27 -- Cc: microscopy-at-msa.microscopy.com 13, 27 -- Message-ID: {4427C88F.2117.818086-at-localhost} 13, 27 -- Priority: normal 13, 27 -- In-reply-to: {200603241938.k2OJc0s8012169-at-ns.microscopy.com} 13, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 13, 27 -- Content-type: text/plain; charset=US-ASCII 13, 27 -- Content-transfer-encoding: 7BIT 13, 27 -- Content-description: Mail message body 13, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
Here are the happy-ending results of your wise advices for TC embedding, both in monolayer and pellet. Both methods worked well, with the advantage of monolayer embedding being that it is 1 day shorter (the penetration times are reduced) and keeps intact the cell contacts. The only disadvantage I found was by cutting, which requires either a lot of experience or a lot of patience (or both?). The cultures must be pretty confluent too, which is an evident limitation. I list the methods I found successful because they may be helpful for somebody else. I don’t want to favor a method for another, I just list what worked for me. This does not mean that other proposed methods are not valid!!
1. For pellet embedding, the good trick was to add the fixative in the culture medium at 37°C, wait 30 sec and scrape the cells, then pellet briefly in eppis at full speed, then replace with fresh fixative and continuing fixation at 4°C. 2. For monolayer embedding, the following method gave me entire satisfaction: do all steps in 3.5 cm petri dish, simply avoiding propyleneoxyde (mix epon with ethanol). Cover the cells with a thin layer of Epon (1 ml/3.5 cm petri dish, which gives approx. the same epon thickness as the bottom of the petri dish) and put BEEM capsules, whose tip has previously been cut out, upside-down in the resin. The next day, the BEEM capsules being embedded in the thin layer of Epon, fill them with Epon and cure for another 24 h. Next day, you can simply pull the capsules out of the petri dishes. It may happen that some plastic comes with the capsule, but never the entire surface, just choose a place without plastic to cut the pyramid.
I want to thank warmly all listers who helped me, and the others too because it wouldn’t be fair otherwise ;-)
Stephane-without-i
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==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Mon Mar 27 03:04:35 2006 7, 18 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2R94ZN9003966 7, 18 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 03:04:35 -0600 7, 18 -- Received: (qmail 42811 invoked by uid 60001); 27 Mar 2006 09:04:35 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=sH4swVF+dgCyC1MJ5E++jtFHS8ZY/AaK2W1ufZY5MjlwnTEF0brCbLkJkz2TCNprhe4aoFUSRqK2H8Aste8wdFbPacSBD42sc2y/u+SL/NmUimpGePNyHdcumYJ5xW+x/9KopnUzGA8aUyRhoazBEZnhWPipN9rpGuj+/e0sATY= ; 7, 18 -- Message-ID: {20060327090435.42807.qmail-at-web37413.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Mon, 27 Mar 2006 01:04:35 PST 7, 18 -- Date: Mon, 27 Mar 2006 01:04:35 -0800 (PST) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: TC cell preparation: happy end and conclusion 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; charset=iso-8859-1 7, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I am looking of an adhesive of very low viscosity that will take onto titanium. The purpose is for the preparation of cross-sectional TEM specimens. The M-bond 610 works well for almost everything else, except Ti. Can someone help me with advice please?.
Thanks
Basil
Dr. Basil Julies Head Electron Microscope Unit Physics Department University of the Western Cape Private Bag X17 Bellville 7535 Tel : (27)(21) 959 2327 or 959 3458 Fax : (27)(21) 959 1335 or 959 3474
==============================Original Headers============================== 5, 19 -- From bjulies-at-uwc.ac.za Mon Mar 27 09:20:26 2006 5, 19 -- Received: from uwcunx.uwc.ac.za (uwcunx.uwc.ac.za [196.11.235.8]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2RFKO9F001403 5, 19 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 09:20:25 -0600 5, 19 -- Received: (qmail 7298 invoked from network); 27 Mar 2006 14:27:13 -0000 5, 19 -- Received: from itsnw.uwc.ac.za (HELO Services-02.uwc.ac.za) (192.102.9.71) 5, 19 -- by uwcunx.uwc.ac.za with SMTP; 27 Mar 2006 14:27:13 -0000 5, 19 -- Received: from UWC-MTA by Services-02.uwc.ac.za 5, 19 -- with Novell_GroupWise; Mon, 27 Mar 2006 17:24:09 +0200 5, 19 -- Message-Id: {s4281fb9.005-at-Services-02.uwc.ac.za} 5, 19 -- X-Mailer: Novell GroupWise Internet Agent 6.5.2 5, 19 -- Date: Mon, 27 Mar 2006 17:23:37 +0200 5, 19 -- From: "Basil Julies" {bjulies-at-uwc.ac.za} 5, 19 -- To: {microscopy-at-microscopy.com} 5, 19 -- Subject: adhesive for Titanium 5, 19 -- Mime-Version: 1.0 5, 19 -- Content-Type: text/plain; charset=US-ASCII 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- Content-Disposition: inline ==============================End of - Headers==============================
First of all I would like to thank everyone for the helpful advice I received earlier on the vapor fixation. The processing of bacteria with paraformaldehyde (3hours) following with 1h of osmium vapor (I tried this combination in reverse as well); vapor dehydration gave me a good fixation as far as I can see at my magnification. Unfortunately, I encountered another problem: salt crystals. By skipping wash I left buffer precipitates seemingly intact yet washing is removing not only salt but also most of the bacteria from the surface treated with antimicrobial. I would greatly appreciate your advice. Albina
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
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Hi Albina we have a number of researchers who use vapour fixation for protists using a protocol from Brian Leander's lab. 4% osmium onto a filter paper in the lid of a petri dish and the living sample in the dish base for 30 minutes. Then one drop of 4% osmium added to the mix per ml of liquid. Leave for 30 minutes and dehydrate as normal after putting the specimens onto a nuclearpore filter in a Millipore Swinnex holder.
The vapor fixation results are superb and several euglenoids from Brian Leander have become magazine cover shots.
We have been trying to use the microwave for the dehydration but have found that the samples are not as sticky as when glutaraldehyde is used first and the samples tended to lift off the nucleopore filter in HMDS. This seems to be less of a problem when we use the critical point dryer. However, we have found that if it works conventionally, it will generally work in the microwave at a fraction of the time if you find the right conditions. This is still a work in progress. It seems that some sample is lost even in the critical point dryer. By leaving the filter in the syringe/swinnex holder all the way through the processing, less is lost even with HMDS in the microwave.
Elaine
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-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada (2003-2005) University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
==============================Original Headers============================== 9, 30 -- From ech-at-interchange.ubc.ca Mon Mar 27 13:40:47 2006 9, 30 -- Received: from mta6.mail-relay.ubc.ca (mta6.mail-relay.ubc.ca [137.82.45.12]) 9, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2RJekFT024817 9, 30 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 13:40:47 -0600 9, 30 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 9, 30 -- by mta6.mail-relay.ubc.ca (8.12.11/8.12.11) with ESMTP id k2RJejfl015773 9, 30 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 11:40:45 -0800 (PST) 9, 30 -- (envelope-from ech-at-interchange.ubc.ca) 9, 30 -- Received: from [24.82.105.155] 9, 30 -- (S01060040f4371335.vn.shawcable.net [24.82.105.155]) 9, 30 -- by smtp.interchange.ubc.ca 9, 30 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 9, 30 -- with ESMTPA id {0IWS00MQZXBW5K-at-smtp.interchange.ubc.ca} for 9, 30 -- microscopy-at-microscopy.com; Mon, 27 Mar 2006 11:40:45 -0800 (PST) 9, 30 -- Date: Mon, 27 Mar 2006 11:40:42 -0800 9, 30 -- From: Elaine Humphrey {ech-at-interchange.ubc.ca} 9, 30 -- Subject: Re: [Microscopy] Part 2: vapor fixation for SEM 9, 30 -- In-reply-to: {200603271800.k2RI0BwK016494-at-ns.microscopy.com} 9, 30 -- X-Sender: ech-at-mail.interchange.ubc.ca 9, 30 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 9, 30 -- Cc: amich-at-ufl.edu 9, 30 -- Message-id: {a06100502c04de6b97f2a-at-[24.82.105.155]} 9, 30 -- MIME-version: 1.0 9, 30 -- Content-type: text/plain; format=flowed; charset=us-ascii 9, 30 -- References: {200603271800.k2RI0BwK016494-at-ns.microscopy.com} 9, 30 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.1.0.0, Antispam-Data: 2006.03.27.105106 9, 30 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 9, 30 -- X-PerlMx-Spam: Probability=7%, Report=IP_HTTP_ADDR 0, __C230066_P1_5 0, __C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0 9, 30 -- X-Spam-Level: 9, 30 -- X-Spam-Flag: No ==============================End of - Headers==============================
thank you for the advice. I will look up Brian Leander's publications. My primary objective is to evaluate changes in the bacteria inhibited by antimicrobial agents. The challenge is working with fibrous substrate inoculated with bacteria to test antimicrobial treatments. In many cases I would like avoid wetting substrates; so vapor fixation is a way to go with the exception of salt residue left behind. Even minimal wash removes bacteria from the treated substrate because they adhesive properties are compromised yet without wash salt crystals are obscuring view. Albina
On Mon Mar 27 14:40:42 EST 2006, Elaine Humphrey {ech-at-interchange.ubc.ca} wrote:
} Hi Albina } we have a number of researchers who use vapour fixation for } protists using a protocol from Brian Leander's lab. 4% osmium } onto a filter paper in the lid of a petri dish and the living } sample in the dish base for 30 minutes. Then one drop of 4% } osmium added to the mix per ml of liquid. Leave for 30 minutes } and dehydrate as normal after putting the specimens onto a } nuclearpore filter in a Millipore Swinnex holder. } } The vapor fixation results are superb and several euglenoids from } Brian Leander have become magazine cover shots. } } We have been trying to use the microwave for the dehydration but } have found that the samples are not as sticky as when } glutaraldehyde is used first and the samples tended to lift off } the nucleopore filter in HMDS. This seems to be less of a problem } when we use the critical point dryer. However, we have found that } if it works conventionally, it will generally work in the } microwave at a fraction of the time if you find the right } conditions. This is still a work in progress. It seems that some } sample is lost even in the critical point dryer. By leaving the } filter in the syringe/swinnex holder all the way through the } processing, less is lost even with HMDS in the microwave. } } Elaine } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } First of all I would like to thank everyone for the helpful } } advice } } I received earlier on the vapor fixation. The processing of } } bacteria with paraformaldehyde (3hours) following with 1h of } } osmium vapor (I tried this combination in reverse as well); vapor } } dehydration gave me a good fixation as far as I can see at my } } magnification. } } Unfortunately, I encountered another problem: salt crystals. By } } skipping wash I left buffer precipitates seemingly intact yet } } washing is removing not only salt but also most of the bacteria } } from the surface treated with antimicrobial. I would greatly } } appreciate your advice. } } Albina } } } } -- } } MIKHAYLOVA,ALBINA, PhD } } Post Doctoral Research Associate } } Materials Science and Engineering } } University of Florida } } } } } } ==============================Original } } Headers============================== } } 3, 21 -- From amich-at-ufl.edu Mon Mar 27 11:58:14 2006 } } 3, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu } } [128.227.74.165]) } } 3, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k2RHwD3v013670 } } 3, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 } } 11:58:14 -0600 } } 3, 21 -- Received: from osgjas01.cns.ufl.edu } } (osgjas01.cns.ufl.edu [128.227.74.131]) } } 3, 21 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id } } k2RHw8kE946362 } } 3, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 } } 12:58:08 -0500 } } 3, 21 -- Message-ID: } } {608177447.7451143482288345.JavaMail.osg-at-osgjas01.cns.ufl.edu} } } 3, 21 -- Date: Mon, 27 Mar 2006 12:58:08 -0500 (EST) } } 3, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} } } 3, 21 -- To: Microscopy-at-microscopy.com } } 3, 21 -- Subject: Part 2: vapor fixation for SEM } } 3, 21 -- MIME-Version: 1.0 } } 3, 21 -- Content-Type: text/plain; format=flowed; } } charset=us-ascii } } 3, 21 -- Content-Transfer-Encoding: 7bit } } 3, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) } } 3, 21 -- X-Originating-IP: 70.152.34.47 [70.152.34.47] } } 3, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED } } 3, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, } } tests=ALL_TRUSTED } } 3, 21 -- X-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } 3, 21 -- X-UFL-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } ==============================End of - } } Headers============================== } } } -- Dr. Elaine Humphrey } Director, BioImaging Facility } President, Microscopy Society of Canada (2003-2005) } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-interchange.ubc.ca } website: www.emlab.ubc.ca } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 8, 22 -- From amich-at-ufl.edu Mon Mar 27 13:57:56 2006 8, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2RJvsxx002025 8, 22 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 13:57:55 -0600 8, 22 -- Received: from osgjas01.cns.ufl.edu (osgjas01.cns.ufl.edu [128.227.74.131]) 8, 22 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k2RJvlVY3825720; 8, 22 -- Mon, 27 Mar 2006 14:57:47 -0500 8, 22 -- Message-ID: {2095729987.15611143489467440.JavaMail.osg-at-osgjas01.cns.ufl.edu} 8, 22 -- Date: Mon, 27 Mar 2006 14:57:47 -0500 (EST) 8, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 8, 22 -- To: Elaine Humphrey {ech-at-interchange.ubc.ca} , 8, 22 -- Microscopy Listserver {microscopy-at-microscopy.com} 8, 22 -- Subject: Re: [Microscopy] Part 2: vapor fixation for SEM 8, 22 -- MIME-Version: 1.0 8, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 8, 22 -- Content-Transfer-Encoding: 7bit 8, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 8, 22 -- X-Originating-IP: 70.152.34.47 [70.152.34.47] 8, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 8, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Dear Basil, Many years ago , when we first started to try to do cross-sections, we used Devcon 2-Ton epoxy for the cross-sections, while we were waiting for the M-610 Bond to arrive. It is not as thin as the M-610, but with pressure on our home-built parallel-jaw clamp the glue joints were thin enough and we got good results. This is a slow, 24 hour cure epoxy. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- X-from: {bjulies-at-uwc.ac.za} To: {mager-at-interchange.ubc.ca} Sent: Monday, March 27, 2006 7:26 AM
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Email: isabeln-at-popsrv.ist.utl.pt Name: Isabel Dias Nogueira
Organization: MicroLab - Instituto Superior Tecnico
Title-Subject: [Filtered] FEG-SEM: cold cathode versus Schottky
Question: Our lab is in the process of buying a FEG-SEM. The main question at this point is whether to choose cold cathode (higher resolution) or Schottky emission (higher current). At first we thought we should go for cold cathode because of the resolution, but we also plan to acquire a EBSD (diffraction using Kikuchi lines) and a EDS detector for mapping, both requiring high beam current. The EBSD, in particular, also requires long acquisition times, which may not work well with the need of flashing the cold cathode emitter every 10h or so. Another issue is current stability: will the lower current stability of the cold cathode (5%) have any consequences on quantifying point EDS analysis ? I would appreciate any comments from users of both types of FEG-SEM, and also of EBSD and EDS mapping. Thanks,
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Email: p.arico-at-email.it Name: Pietro Arico'
Organization: Dept. CFTA - University of Palermo (IT)
Title-Subject: [Filtered] EDS Co Calibration
Question: dear all, i am trying to acquire spectra of silicate and metallic standards using a LEO 440 coupled with an Oxford Link Isis 300 EDS system. I have acquired 37 spectra of different standards doing a calibration using Cobalt to check the instrument stability (I have no Faraday cup and Current meter). Here are the values of the calibrations: Time 14.15 14.25 14.35 14.45 14.55 15.05 15.35 16.15 16.35 16.55 17.15 Zero energy channel 9.623245 9.622585 9.619363 9.624291 9.623804 9.626814 9.623498 9.623422 9.623599 9.624767 9.624976 Energy per channel (eV) 20.00182 19.99826 20.00041 20.00109 19.99679 19.99775 19.99981 19.99792 20.00136 20.00056 19.99934 Counts in calibration peak 48880 48736 48457 48225 47876 48331 49103 48991 49426 49188 48925
do you think the instrument is enough stable or not? the differences between these values are negligible or not? thank you very much fopr your help (it's the first time I try to do this!!!)
Pietro Arico' Dept. Chemistry and Physics of the Earth (CFTA) University of Palermo Via Archirafi, 36 Palermo, 90123 - Italy email: p.arico-at-email.it
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Email: ycn1-at-psu.edu Name: Yuk-Chow Ng
Organization: Penn State University
Title-Subject: [Filtered] quantitation of immunofluorescence
Question: I am trying to perform a semi-quantitative comparison of immunofluorescence intensity on the sarcoplasmic membrane of skeletal muscle fibers (cross sections). Is there a way to trace the signals on the membrane of multiple fibers and compare the overall intensity between a control and a treated sample (sections).
The calibration of EDS is usually done at Al and Cu.
I use X-Checker Extra to do this since it does Al and Cu plus F, Be, C, B, N.
The first pass is Al+Cu. Then separate passes for lower Z elements are lower KV.
Disclaimer: I have no financial interest in X-checker.
gary g.
At 04:55 PM 3/27/2006, you wrote:
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==============================Original Headers============================== 12, 20 -- From gary-at-gaugler.com Mon Mar 27 19:16:55 2006 12, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S1GqRr022289 12, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 19:16:53 -0600 12, 20 -- Received: (qmail 12667 invoked from network); 27 Mar 2006 17:16:50 -0800 12, 20 -- Received: by simscan 1.1.0 ppid: 12664, pid: 12665, t: 0.1680s 12, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 20 -- by qsmtp3 with SMTP; 27 Mar 2006 17:16:50 -0800 12, 20 -- Message-Id: {7.0.1.0.2.20060327171410.026a6928-at-gaugler.com} 12, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 20 -- Date: Mon, 27 Mar 2006 17:16:51 -0800 12, 20 -- To: p.arico-at-email.it 12, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: EDS Co Calibration 12, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 20 -- In-Reply-To: {200603280055.k2S0tCFc003300-at-ns.microscopy.com} 12, 20 -- References: {200603280055.k2S0tCFc003300-at-ns.microscopy.com} 12, 20 -- Mime-Version: 1.0 12, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Well, this is a significant decision point indeed.
I don't necessarily agree that cold cathode FE SEMs produce higher resolution. But that is not the issue here.
Schottky is going to produce a very high current beam with astounding stability. This is nice but critical for EBSD. However, what is the maximum probe current available? You can have great stability of a low current system and spend hours on an EBSD scan. You also have to figure out which EBSD system you are going to use. TSL offers drift correction (good at } 1KX) while AFIK, HKL does not offer. These are your main two providers of EBSD.
Your beam strength will greatly impact the fps of the EBSD data collection. fps of between 22-33 are good. Higher fps may be dependent on degradation of probe diameter. So watch out for this.
Now, throwing in EDS mapping you are moving another step. These maps can take hours to complete--depending on cps and DT. Either way, the Schottky FE is going to be superior, IMO.
Bottom line...get a Schottky FE with as high a probe current as you can get with adjustable probe diameters. BTW, each SEM maker does this differently. Most use final apertures. Zeiss does not.
Contact me off-line if you would like some specifics.
Disclaimer: No financial interest in any supplier I reference.
gary g.
At 04:55 PM 3/27/2006, you wrote:
} This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form } at http://microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both isabeln-at-popsrv.ist.utl.pt as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: isabeln-at-popsrv.ist.utl.pt } Name: Isabel Dias Nogueira } } Organization: MicroLab - Instituto Superior Tecnico } } Title-Subject: [Filtered] FEG-SEM: cold cathode versus Schottky } } Question: Our lab is in the process of buying a FEG-SEM. } The main question at this point is whether to choose cold cathode } (higher resolution) or Schottky emission (higher current). At first } we thought we should go for cold cathode because of the resolution, } but we also plan to acquire a EBSD (diffraction using Kikuchi lines) } and a EDS detector for mapping, both requiring high beam current. } The EBSD, in particular, also requires long acquisition times, which } may not work well with the need of flashing the cold cathode emitter } every 10h or so. Another issue is current stability: will the lower } current stability of the cold cathode (5%) have any consequences on } quantifying point EDS analysis ? I would appreciate any comments } from users of both types of FEG-SEM, and also of EBSD and EDS mapping. Thanks, } } Isabel } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 7, 12 -- From zaluzec-at-microscopy.com Mon Mar 27 18:52:08 2006 } 7, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 7, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k2S0q8n0026146 } 7, 12 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 } 18:52:08 -0600 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: (Unverified) } 7, 12 -- Message-Id: {p06110407c04e3929fcf5-at-[206.69.208.22]} } 7, 12 -- Date: Mon, 27 Mar 2006 18:52:06 -0600 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: isabeln-at-mail.ist.utl.pt (by way of MicroscopyListserver) } 7, 12 -- Subject: viaWWW: FEG-SEM: cold cathode versus Schottky } 7, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
==============================Original Headers============================== 14, 20 -- From gary-at-gaugler.com Mon Mar 27 19:29:38 2006 14, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S1Tceo031943 14, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Mar 2006 19:29:38 -0600 14, 20 -- Received: (qmail 987 invoked from network); 27 Mar 2006 17:28:55 -0800 14, 20 -- Received: by simscan 1.1.0 ppid: 981, pid: 983, t: 0.0845s 14, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 14, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 14, 20 -- by qsmtp4 with SMTP; 27 Mar 2006 17:28:54 -0800 14, 20 -- Message-Id: {7.0.1.0.2.20060327171705.026b7008-at-gaugler.com} 14, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 20 -- Date: Mon, 27 Mar 2006 17:29:38 -0800 14, 20 -- To: isabeln-at-mail.ist.utl.pt 14, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 14, 20 -- Subject: Re: [Microscopy] viaWWW: FEG-SEM: cold cathode versus Schottky 14, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 14, 20 -- In-Reply-To: {200603280055.k2S0tCiJ003290-at-ns.microscopy.com} 14, 20 -- References: {200603280055.k2S0tCiJ003290-at-ns.microscopy.com} 14, 20 -- Mime-Version: 1.0 14, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
First of all be careful when taking about signal intensity in fluorescence, especially immunofluorescence, since the amount of signal is not directly related to the concentration of the target. You may however compare the intensity of 2 samples treated exactly the same way, still being careful not to draw too precise conclusions. Microscopy is not the method of choice for quantification. For your special need, I suppose that using a confocal microscope you could draw a profile of fluorescence or define a ROI (region of interest) and let the computer calculate the amount of fluorescence. Don't forget to use the same settings for all samples otherwise no comparison is possible! (manipulation of signal is so easy on a confocal!)
Stephane-without-i
--- ycn1-at-psu.edu wrote:
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Tue Mar 28 00:31:16 2006 9, 20 -- Received: from web37401.mail.mud.yahoo.com (web37401.mail.mud.yahoo.com [209.191.87.54]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S6VG8g012752 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 00:31:16 -0600 9, 20 -- Received: (qmail 24141 invoked by uid 60001); 28 Mar 2006 06:31:15 -0000 9, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 20 -- s=s1024; d=yahoo.com; 9, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 20 -- b=nuWJ184L0ZuIRfCk4NQVTL/5DM1hFCHLEt4345+F3T0KpaNJGXrKBvp0I7X3zFAmIr0Y8o7NqY/GrcEt7FaqEOONq999+UYTHM/8zgtmpCE++HkveAay0WwToTVDeuk/Yvm6u7o22XeU/I1VOiOwOg3x2cjFwH127a2GqqcQ7Os= ; 9, 20 -- Message-ID: {20060328063115.24139.qmail-at-web37401.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37401.mail.mud.yahoo.com via HTTP; Mon, 27 Mar 2006 22:31:15 PST 9, 20 -- Date: Mon, 27 Mar 2006 22:31:15 -0800 (PST) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] viaWWW: quantitation of immunofluorescence 9, 20 -- To: ycn1-at-psu.edu 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200603280101.k2S11SjV020852-at-ns.microscopy.com} 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; charset=iso-8859-1 9, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I have never made a detailed study about superglue outgassing, but when used as a crevice and pore back-fill on a polished specimens (and suitably cured) I have not noticed a vacuum problem. This is in the area of 5E-5 to 5E-6 Torr. This is not to say it does not outgas, but only that my pumping system has no trouble keeping up.
OTOH, it is NOT beam stable. I have seen it boil under the beam at anything less than very low kV and current.
Regards, Woody White BWXT Services
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Dear Colleagues,
I would appreciate it if some of you could give me your experience of how the widely available Superglue behaves under vacuum in the SEM and TEM.
Is it a suitable adhesive?
Is there too much de-gassing etc.
Many thanks,
Ted Dunn The EMscope Company Ltd.
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==============================Original Headers============================== 10, 19 -- From drteddunne-at-yahoo.com Tue Mar 28 03:45:05 2006 10, 19 -- Received: from web33407.mail.mud.yahoo.com (web33407.mail.mud.yahoo.com [68.142.206.139]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S9j4Qn026844 10, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 03:45:04 -0600 10, 19 -- Received: (qmail 73056 invoked by uid 60001); 28 Mar 2006 09:45:04 -0000 10, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 19 -- s=s1024; d=yahoo.com; 10, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Cont ent-Type:Content-Transfer-Encoding; 10, 19 -- b=qc0c3Nz3PGKgGIW9L3FWLolqeL2G4o9CtPkFC+5Zckimur+GWWIvWx9YYEy7qpRMWlAQAq QAbcOIj7cNINrfdTJti374bhqp+etRhqMY3hXdHKg7AaggEpXQCX3TvlXm+kRRetbpQOK3ix qg58Ocryz8g0IcMkqPmgjr/RKr9JQ= ; 10, 19 -- Message-ID: {20060328094504.73054.qmail-at-web33407.mail.mud.yahoo.com} 10, 19 -- Received: from [202.47.247.116] by web33407.mail.mud.yahoo.com via HTTP; Tue, 28 Mar 2006 01:45:04 PST 10, 19 -- Date: Tue, 28 Mar 2006 01:45:04 -0800 (PST) 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 19 -- Subject: Superglue 10, 19 -- To: microscopy-at-microscopy.com 10, 19 -- In-Reply-To: {200603272125.k2RLPJn3019014-at-ns.microscopy.com} 10, 19 -- MIME-Version: 1.0 10, 19 -- Content-Type: text/plain; charset=iso-8859-1 10, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 19, 27 -- From nwwhite-at-bwxt.com Tue Mar 28 07:04:34 2006 19, 27 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 19, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SD4XQC016566 19, 27 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Mar 2006 07:04:33 -0600 19, 27 -- Received: from ([131.184.13.224]) 19, 27 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.545394; 19, 27 -- Tue, 28 Mar 2006 08:04:20 -0500 19, 27 -- Received: from bwxslynpo01.BWXS.BWXTECH.NET ([131.184.13.4]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 19, 27 -- Tue, 28 Mar 2006 08:04:19 -0500 19, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 19, 27 -- Content-class: urn:content-classes:message 19, 27 -- MIME-Version: 1.0 19, 27 -- Content-Type: text/plain; 19, 27 -- charset="US-ASCII" 19, 27 -- Subject: RE: [Microscopy] Superglue 19, 27 -- Date: Tue, 28 Mar 2006 08:04:19 -0500 19, 27 -- Message-ID: {70E4EA352EC987489773D55B0FF83E1C1DF9C4-at-bwxslynpo01.BWXS.BWXTECH.NET} 19, 27 -- X-MS-Has-Attach: 19, 27 -- X-MS-TNEF-Correlator: 19, 27 -- Thread-Topic: [Microscopy] Superglue 19, 27 -- Thread-Index: AcZSTH+0uE/ojL+RRHKVxLWBujyaFgAGnfog 19, 27 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 19, 27 -- To: {drteddunne-at-yahoo.com} , 19, 27 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 19, 27 -- X-OriginalArrivalTime: 28 Mar 2006 13:04:19.0972 (UTC) FILETIME=[205D9C40:01C65268] 19, 27 -- Content-Transfer-Encoding: 8bit 19, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2SD4XQC016566 ==============================End of - Headers==============================
I am posting message for Dr. Milos Kalab who is not a subscriber. His web pages have been recommended from time to time on the list.
His messages are as following: Milos Kalab, Honorary Research Associate at Agriculture and Agri-Food Canada in Ottawa, who has many websites on electron microscopy of foods and microorganisms hosted by the generosity of the University of Lund in Sweden on their server with URL either http://distans.livstek.lth.se:2080/ or http://anka.livstek.lth.se:2080/ wants to apologize to their visitors. The server in Sweden has been out of order and it is not known when it will be repaired. The starting point with links to those websites is in Canada at http://www.magma.ca/~scimat/ where new information may be found. Milos has been transferring some of the websites to active addresses to make them accessible again. He may be contacted at scimat-at-magma.ca . Thank you.
Ann Fook Yang Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 Room 2097, K.W. Neatby Bldg., CEF , Ottawa, ON, Canada K1A 0C6 yanga-at-agr.gc.ca
==============================Original Headers============================== 3, 26 -- From YANGA-at-AGR.GC.CA Tue Mar 28 09:22:55 2006 3, 26 -- Received: from agrpazsmtp2.agr.gc.ca (agrpazsmtp2.agr.gc.ca [192.197.71.137]) 3, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFMtF3031662 3, 26 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 09:22:55 -0600 3, 26 -- Received: from onncrxcn1.AGR.GC.CA ([192.168.122.1]) 3, 26 -- by agrpazsmtp2.agr.gc.ca (8.13.6/8.13.3) with ESMTP id k2SFJ0mu023222 3, 26 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:19:00 -0500 3, 26 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn1.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 3, 26 -- Tue, 28 Mar 2006 10:22:55 -0500 3, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 3, 26 -- content-class: urn:content-classes:message 3, 26 -- MIME-Version: 1.0 3, 26 -- Content-Type: text/plain; 3, 26 -- charset="iso-8859-1" 3, 26 -- Subject: web problem 3, 26 -- Date: Tue, 28 Mar 2006 10:22:54 -0500 3, 26 -- Message-ID: {E035A9C87303AE4AB9BA10FD8324DFB1024334B6-at-onncrxms3.agr.gc.ca} 3, 26 -- X-MS-Has-Attach: 3, 26 -- X-MS-TNEF-Correlator: 3, 26 -- Thread-Topic: web problem 3, 26 -- Thread-Index: AcZR2hZo/1G+jdJlQ9e2aCztWikm7Q== 3, 26 -- From: "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 3, 26 -- To: {microscopy-at-microscopy.com} 3, 26 -- X-OriginalArrivalTime: 28 Mar 2006 15:22:55.0132 (UTC) FILETIME=[7C9655C0:01C6527B] 3, 26 -- Content-Transfer-Encoding: 8bit 3, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2SFMtF3031662 ==============================End of - Headers==============================
Just a short note to thanks everyone who e-mailed or called me to provide assistance with the identification of titanium from titanium oxide in an organic binder. Your assistance was much appreciated.
If your interested, I ashed my sample to reduce the organic fraction and fluxed the remaining material in sodium borate with a platinum wire and alcohol lamp (How many labs still have alcohol lamps…). The bead was removed, crushed and dissolved in concentrated H2SO4. Both the hydrogen peroxide and chromotropic acid tests as described as per Feigl in “Qualitative Analysis of Spot Tests” were used. I tried both tests as a spot test on filter paper, in a capillary tube and in a white spot plate. The spot test worked best for me.
Simply heating a sample of TiO2 in concentrated H2SO4 did not convert much of the relatively inert TiO2 into a detectable form, but fluxing did.
Thanks again!!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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==============================Original Headers============================== 10, 19 -- From frank.karl-at-degussa.com Tue Mar 28 09:38:42 2006 10, 19 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFcgoh008854 10, 19 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Mar 2006 09:38:42 -0600 10, 19 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 10, 19 -- by framailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k2SFcdoH024564 10, 19 -- for {microscopy-at-msa.microscopy.com} ; Tue, 28 Mar 2006 17:38:40 +0200 10, 19 -- Subject: Yes, my sample contained titanium 10, 19 -- To: microscopy-at-msa.microscopy.com 10, 19 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 10, 19 -- Message-ID: {OF668E6ACA.056DB750-ON8525713F.0055C11D-8525713F.0055EC98-at-degussa.com} 10, 19 -- From: frank.karl-at-degussa.com 10, 19 -- Date: Tue, 28 Mar 2006 10:38:31 -0500 10, 19 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 10, 19 -- 03/28/2006 09:38:41 AM 10, 19 -- MIME-Version: 1.0 10, 19 -- Content-type: text/plain; charset=UTF-8 10, 19 -- Content-Transfer-Encoding: 8bit 10, 19 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k2SFcgoh008854 ==============================End of - Headers==============================
I use Gatan's G-1 epoxy for all my cross sections. (You can get the same product from Epo-tek as well, I believe they are the original manufacturer). I have not tried it with Ti substrates, but I routinely do silicon, glass, MgO, SrTiO3, etc. I wouldn't call it low viscosity, but I regularly get glue lines less than 10 nm as measured in the TEM. I cure it on a hotplate around 100C in a vice with as much pressure as I can get. Hope that helps!
Leslie
Leslie Krupp (Thompson) Engineer/Scientiest IBM Almaden Research 650 Harry Road, K19/D2 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 4, 27 -- From lkrupp-at-us.ibm.com Tue Mar 28 09:44:33 2006 4, 27 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) 4, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFiX1O018381 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 09:44:33 -0600 4, 27 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 4, 27 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k2SFiWIs016723 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:33 -0500 4, 27 -- Received: from d01av01.pok.ibm.com (d01av01.pok.ibm.com [9.56.224.215]) 4, 27 -- by d01relay04.pok.ibm.com (8.12.10/NCO/VER6.8) with ESMTP id k2SFiMFj176842 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:22 -0500 4, 27 -- Received: from d01av01.pok.ibm.com (loopback [127.0.0.1]) 4, 27 -- by d01av01.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k2SFiMvs012415 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:22 -0500 4, 27 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 4, 27 -- by d01av01.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k2SFiM2N012390 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:44:22 -0500 4, 27 -- To: microscopy-at-microscopy.com 4, 27 -- MIME-Version: 1.0 4, 27 -- Subject: Re: Adehsive for Titanium 4, 27 -- X-Mailer: Lotus Notes Release 6.0.2CF1 June 9, 2003 4, 27 -- Message-ID: {OF017C7D97.579B9454-ON8525713F.0055E41F-8825713F.00567582-at-us.ibm.com} 4, 27 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 4, 27 -- Date: Tue, 28 Mar 2006 07:44:21 -0800 4, 27 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF18 | February 28, 2006) at 4, 27 -- 03/28/2006 10:44:22, 4, 27 -- Serialize complete at 03/28/2006 10:44:22 4, 27 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
While outgassing may not be a problem I have frequently had issues with bond failure when exposed to temperature excursions. Thus I do not use superglue for anything I want permanently bonded.
Much better are the epoxies, such as Gatan's G-1 or the M-Bond brand. These are stable under the e-beam and do not outgas.
-- Roger A. Ristau, PhD TEM Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269 vox: 860-486-5379 fax: 860-486-4745
==============================Original Headers============================== 6, 19 -- From raristau-at-ims.uconn.edu Tue Mar 28 09:54:58 2006 6, 19 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SFsvkG027967 6, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 09:54:58 -0600 6, 19 -- Received: from [137.99.44.251] (d44h251.public.uconn.edu [137.99.44.251]) 6, 19 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k2SFsZXC022431 6, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 10:54:35 -0500 6, 19 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 19 -- Date: Tue, 28 Mar 2006 10:54:18 -0500 6, 19 -- Subject: re: Superglue 6, 19 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 6, 19 -- To: {microscopy-at-microscopy.com} 6, 19 -- Message-ID: {C04EC65A.AD8%raristau-at-ims.uconn.edu} 6, 19 -- Mime-version: 1.0 6, 19 -- Content-type: text/plain; charset="US-ASCII" 6, 19 -- Content-transfer-encoding: 7bit 6, 19 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 6, 19 -- X-UConn-MailScanner: Found to be clean 6, 19 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
It's not a micro-chemical test, but TAPPI Test Method T627 "Determination of Titanium dioxide" uses ammonium sulfate in sulfuric acid. They also list an XRF test, method T554. Methods can be purchased individually from www.tappi.org.
I have no financial intesest in TAPPI, a paper industry association.
David Rothbard Bureau of Engraving and Printing
frank.karl-at-degussa.com wrote:
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==============================Original Headers============================== 11, 20 -- From rothbardd-at-netscape.net Tue Mar 28 11:38:47 2006 11, 20 -- Received: from imo-d01.mx.aol.com (imo-d01.mx.aol.com [205.188.157.33]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SHckpL006710 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 11:38:46 -0600 11, 20 -- Received: from rothbardd-at-netscape.net 11, 20 -- by imo-d01.mx.aol.com (mail_out_v38_r7.3.) id w.13b.13377e91 (16237) 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 12:38:42 -0500 (EST) 11, 20 -- Received: from netscape.net (mow-d20.webmail.aol.com [205.188.139.161]) by air-in03.mx.aol.com (v108_r3.6) with ESMTP id MAILININ31-3f6d442974a2233; Tue, 28 Mar 2006 12:38:42 -0500 11, 20 -- Date: Tue, 28 Mar 2006 12:38:42 -0500 11, 20 -- From: rothbardd-at-netscape.net 11, 20 -- To: microscopy-at-microscopy.com 11, 20 -- Subject: RE: Question Microchemical test for Ti 11, 20 -- MIME-Version: 1.0 11, 20 -- Message-ID: {38C1CBEB.7C245EF9.034D9A6A-at-netscape.net} 11, 20 -- X-Mailer: Atlas Mailer 2.0 11, 20 -- X-AOL-IP: 199.196.144.13 11, 20 -- X-AOL-Language: english 11, 20 -- Content-Type: text/plain; charset=iso-8859-1 11, 20 -- Content-Transfer-Encoding: 8bit 11, 20 -- X-Spam-Flag: NO ==============================End of - Headers==============================
Ted: I've used "Super Glue" (the Loctite variety, mostly) for years to mount samples for cross-sectioning and then imaging in the SEM, both thermionic and cold FE types. I've had no problems with outgassing, even with my JEOL 6600FXV, which is very sensitive to any outgassing.
drteddunne-at-yahoo.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Colleagues, } } I would appreciate it if some of you could give me } your experience of how the widely available Superglue } behaves under vacuum in the SEM and TEM. } } Is it a suitable adhesive? } } Is there too much de-gassing etc. } } Many thanks, } } } Ted Dunn } The EMscope Company Ltd. } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 10, 19 -- From drteddunne-at-yahoo.com Tue Mar 28 03:45:05 2006 } 10, 19 -- Received: from web33407.mail.mud.yahoo.com (web33407.mail.mud.yahoo.com [68.142.206.139]) } 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S9j4Qn026844 } 10, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 03:45:04 -0600 } 10, 19 -- Received: (qmail 73056 invoked by uid 60001); 28 Mar 2006 09:45:04 -0000 } 10, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 10, 19 -- s=s1024; d=yahoo.com; } 10, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 10, 19 -- b=qc0c3Nz3PGKgGIW9L3FWLolqeL2G4o9CtPkFC+5Zckimur+GWWIvWx9YYEy7qpRMWlAQAqQAbcOIj7cNINrfdTJti374bhqp+etRhqMY3hXdHKg7AaggEpXQCX3TvlXm+kRRetbpQOK3ixqg58Ocryz8g0IcMkqPmgjr/RKr9JQ= ; } 10, 19 -- Message-ID: {20060328094504.73054.qmail-at-web33407.mail.mud.yahoo.com} } 10, 19 -- Received: from [202.47.247.116] by web33407.mail.mud.yahoo.com via HTTP; Tue, 28 Mar 2006 01:45:04 PST } 10, 19 -- Date: Tue, 28 Mar 2006 01:45:04 -0800 (PST) } 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} } 10, 19 -- Subject: Superglue } 10, 19 -- To: microscopy-at-microscopy.com } 10, 19 -- In-Reply-To: {200603272125.k2RLPJn3019014-at-ns.microscopy.com} } 10, 19 -- MIME-Version: 1.0 } 10, 19 -- Content-Type: text/plain; charset=iso-8859-1 } 10, 19 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers============================== } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 22 -- From r-holdford-at-ti.com Tue Mar 28 14:01:56 2006 4, 22 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SK1tsJ018441 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 28 Mar 2006 14:01:55 -0600 4, 22 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 22 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k2SK1jHk027781; 4, 22 -- Tue, 28 Mar 2006 14:01:55 -0600 (CST) 4, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 22 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k2SK1jqq026509; 4, 22 -- Tue, 28 Mar 2006 14:01:45 -0600 (CST) 4, 22 -- Message-ID: {44299628.5050003-at-ti.com} 4, 22 -- Date: Tue, 28 Mar 2006 14:01:44 -0600 4, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 22 -- Organization: SC Packaging Development -- FA Development 4, 22 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 4, 22 -- MIME-Version: 1.0 4, 22 -- To: drteddunne-at-yahoo.com, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Re: [Microscopy] Superglue 4, 22 -- References: {200603280945.k2S9jRoG027186-at-ns.microscopy.com} 4, 22 -- In-Reply-To: {200603280945.k2S9jRoG027186-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I've also used Loctite 460 to attach samples to a copper grid for the TEM (using the least amount of glue possible). Prior to putting in the TEM, I cure the glue in our lab vacuum desiccator for ~1 hour. Again, never had a problem.
-----Original Message----- X-from: drteddunne-at-yahoo.com [mailto:drteddunne-at-yahoo.com] Sent: Tuesday, March 28, 2006 3:51 AM To: wgstratton-at-wisc.edu
Dear Colleagues,
I would appreciate it if some of you could give me your experience of how the widely available Superglue behaves under vacuum in the SEM and TEM.
Is it a suitable adhesive?
Is there too much de-gassing etc.
Many thanks,
Ted Dunn The EMscope Company Ltd.
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 10, 19 -- From drteddunne-at-yahoo.com Tue Mar 28 03:45:05 2006 10, 19 -- Received: from web33407.mail.mud.yahoo.com (web33407.mail.mud.yahoo.com [68.142.206.139]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k2S9j4Qn026844 10, 19 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 03:45:04 -0600 10, 19 -- Received: (qmail 73056 invoked by uid 60001); 28 Mar 2006 09:45:04 -0000 10, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 19 -- s=s1024; d=yahoo.com; 10, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Cont ent-Type:Content-Transfer-Encoding; 10, 19 -- b=qc0c3Nz3PGKgGIW9L3FWLolqeL2G4o9CtPkFC+5Zckimur+GWWIvWx9YYEy7qpRMWlAQAq QAbcOIj7cNINrfdTJti374bhqp+etRhqMY3hXdHKg7AaggEpXQCX3TvlXm+kRRetbpQOK3ix qg58Ocryz8g0IcMkqPmgjr/RKr9JQ= ; 10, 19 -- Message-ID: {20060328094504.73054.qmail-at-web33407.mail.mud.yahoo.com} 10, 19 -- Received: from [202.47.247.116] by web33407.mail.mud.yahoo.com via HTTP; Tue, 28 Mar 2006 01:45:04 PST 10, 19 -- Date: Tue, 28 Mar 2006 01:45:04 -0800 (PST) 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 19 -- Subject: Superglue 10, 19 -- To: microscopy-at-microscopy.com 10, 19 -- In-Reply-To: {200603272125.k2RLPJn3019014-at-ns.microscopy.com} 10, 19 -- MIME-Version: 1.0 10, 19 -- Content-Type: text/plain; charset=iso-8859-1 10, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 18, 24 -- From wgstratton-at-wisc.edu Tue Mar 28 14:31:54 2006 18, 24 -- Received: from mail.cae.wisc.edu (mail.cae.wisc.edu [144.92.13.31]) 18, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SKVsak028699 18, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 14:31:54 -0600 18, 24 -- Received: from trogdor (trogdor.msae.wisc.edu [128.104.200.76]) 18, 24 -- by mail.cae.wisc.edu (8.13.3+Sun/8.13.4) with ESMTP id k2SKVqEM009719; 18, 24 -- Tue, 28 Mar 2006 14:31:52 -0600 (CST) 18, 24 -- From: "William Stratton" {wgstratton-at-wisc.edu} 18, 24 -- To: {drteddunne-at-yahoo.com} , {Microscopy-at-microscopy.com} 18, 24 -- Subject: RE: [Microscopy] Superglue 18, 24 -- Date: Tue, 28 Mar 2006 14:31:46 -0600 18, 24 -- Message-ID: {000c01c652a6$a748c250$4cc86880-at-trogdor} 18, 24 -- MIME-Version: 1.0 18, 24 -- Content-Type: text/plain; 18, 24 -- charset="us-ascii" 18, 24 -- Content-Transfer-Encoding: 7bit 18, 24 -- X-Priority: 3 (Normal) 18, 24 -- X-MSMail-Priority: Normal 18, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 18, 24 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2527 18, 24 -- Importance: Normal 18, 24 -- In-Reply-To: {200603280950.k2S9oYIc031851-at-ns.microscopy.com} 18, 24 -- X-CAE-MailScanner-Information: Please contact security-at-engr.wisc.edu if this message contains a virus or has been corrupted in delivery. 18, 24 -- X-CAE-MailScanner: Found to be clean (benji) ==============================End of - Headers==============================
yes, I used G-1 epoxy from Gatan. I usually leave it in air for about 10minutes before heating/curing the epoxy in oven, so I can have very thin glue line.
I am pretty sure that G-1 epoxy works for Ti. I have prapared some bio-samples grown on Ti-substrates. It worked very well. My colleague also used the G-1 epoxy to prepare the samples (they put samples in a Ti-clip supplied by a Hungary comapny, I can nor remember the name), it works well.
---- Original message ---- } Date: Tue, 28 Mar 2006 09:45:25 -0600 } From: lkrupp-at-us.ibm.com } Subject: [Microscopy] Re: Adehsive for Titanium } To: clei-at-uiuc.edu
} Basil- } } I use Gatan's G-1 epoxy for all my cross sections. (You can get the same } product from Epo-tek as well, I believe they are the original } manufacturer). I have not tried it with Ti substrates, but I routinely do } silicon, glass, MgO, SrTiO3, etc. I wouldn't call it low viscosity, but I } regularly get glue lines less than 10 nm as measured in the TEM. I cure } it on a hotplate around 100C in a vice with as much pressure as I can get. } Hope that helps! } } Leslie } } Leslie Krupp (Thompson) } Engineer/Scientiest } IBM Almaden Research } 650 Harry Road, K19/D2 } San Jose, CA 95120-6099 } (408) 927-3856 } } ==============================Original Changhui LEI ******************************* Research Electron Microscopist Center for Microanalysis of Materials Frederick Seitz Materials Research Laboratory 104 S. Goodwin Avenue Urbana, IL 61801 USA email: clei-at-uiuc.edu tel: 1-217-244-6177 fax: 1-217-244-2178
==============================Original Headers============================== 6, 26 -- From clei-at-uiuc.edu Tue Mar 28 15:52:40 2006 6, 26 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu [128.174.5.96]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2SLqeOK006831 6, 26 -- for {microscopy-at-microscopy.com} ; Tue, 28 Mar 2006 15:52:40 -0600 6, 26 -- Received: from expms6.cites.uiuc.edu (expms6.cites.uiuc.edu [128.174.5.43]) 6, 26 -- by expredir5.cites.uiuc.edu (8.13.6/8.13.6) with ESMTP id k2SLqdVP013360; 6, 26 -- Tue, 28 Mar 2006 15:52:39 -0600 (CST) 6, 26 -- Received: from expms6.cites.uiuc.edu (localhost.cites.uiuc.edu [127.0.0.1]) 6, 26 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 6, 26 -- with ESMTP id BGR26023; 6, 26 -- Tue, 28 Mar 2006 15:52:39 -0600 (CST) 6, 26 -- Received: from 128.174.5.212 6, 26 -- by expms6.cites.uiuc.edu (MOS 3.4.8-GR) 6, 26 -- with HTTP/1.1; 6, 26 -- Tue, 28 Mar 2006 14:52:38 -0700 6, 26 -- Date: Tue, 28 Mar 2006 14:52:38 -0700 6, 26 -- From: Changhui LEI {clei-at-uiuc.edu} 6, 26 -- Subject: Re: [Microscopy] Re: Adehsive for Titanium 6, 26 -- To: lkrupp-at-us.ibm.com 6, 26 -- Cc: microscopy-at-microscopy.com 6, 26 -- Reply-To: clei-at-uiuc.edu 6, 26 -- X-Mailer: Webmail Mirapoint Direct 3.4.8-GR 6, 26 -- MIME-Version: 1.0 6, 26 -- Message-Id: {1c12a1f6.a048ceea.8238100-at-expms6.cites.uiuc.edu} 6, 26 -- Content-Type: text/plain; charset=us-ascii 6, 26 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I assume that the time is in hours and that the notation hours.minutes. If so, over a three hour period, your current has varied within a couple percent of its initial value. The "worst" changes came in the first forty minutes, from 14.15 to 14.55, where the intensity fell by 2.1%. After that, it seems that you did something to optimize the column/filament because the intensity came back up and stabilized to within one percent.
Is that good enough for your analyses? I don't know. It probably depends on the measurements you intend to make. It should be good enough for standards. You can go back and re-perform a "quant calibration" more often as you collect standards. That way you should have a good intensity reference for each standard even without a Faraday cup. (For non-ISIS users, I understand that Oxford chose cobalt as a reference material since it was readily available, had K and L peaks detectable in the spectrum, and was not readily subject to oxidation. A "quant calibration" is performed prior to analysis to provide a reference of beam intensity across sessions.)
Another question to consider is that of peak integrals. You have almost 50,000 counts in a peak for a pure element. The standard deviation in that count is 223 or 0.44%, relative. That is roughly on the same order of your beam stability.
However, consider 5% cobalt in an alloy matrix. Your net integral would be 2500 counts with a standard deviation of 50 counts or 2%, relative. That indicates more variability will come from your finite counts than from beam variation. There are also issues of background count.
More detailed and thorough treatments of the error are in the texts on microbeam analysis. However, those texts cannot answer what level of precision or accuracy you require for your analyses.
There are many other issues to be considered before embarking on quantitative analyses. I claim to have performed decent quantitative analysis for years by EDS. Still, I ran into difficulty a few years ago when trying to prepare my own standards to complement our commercially obtained standards. I coated my own standards with carbon and found my totals were off by a couple of percent. It turned out that I had different thicknesses of carbon on my commercial and homemade standards. It made a measurable difference. There are many other issues as well. Be sure to internally check the results of your work to develop confidence in your results.
Warren Straszheim
-----Original Message----- X-from: p.arico-at-email.it [mailto:p.arico-at-email.it] Sent: Monday, March 27, 2006 6:54 PM To: wesaia-at-iastate.edu
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 likun-at-charteredsemi.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: likun-at-charteredsemi.com Name: Simon
Organization: Chartered Semicondcutor
Title-Subject: [Filtered] Lift-out Glass rods
Question: Dear Listeres,
Does anyone of you know the suppliers of lift-out galss rods (1.0 mm in dimameter and solid type)? It will be appreciated if you could kindly share the information.
Free to a good home: LKB Histo Knife maker # 2078. I assume that this is a Ralph knife maker. Some glass too. Very heavy. Yours for he cost of shipping.
Greg
-- Gregory W. Erdos, Ph.D, Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 4, 22 -- From gwe-at-ufl.edu Wed Mar 29 08:56:01 2006 4, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2TEu1wL016916 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Mar 2006 08:56:01 -0600 4, 22 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 4, 22 -- (authenticated bits=0) 4, 22 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k2TEtxvL2998398 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Mar 2006 09:55:59 -0500 4, 22 -- Message-ID: {442A9FFE.5080603-at-ufl.edu} 4, 22 -- Date: Wed, 29 Mar 2006 09:55:58 -0500 4, 22 -- From: Greg Erdos {gwe-at-ufl.edu} 4, 22 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 4, 22 -- X-Accept-Language: en-us, en 4, 22 -- MIME-Version: 1.0 4, 22 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Ralph knife maker 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
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Email: dgmorgan-at-ucdavis.edu Name: David Morgan
Organization: UC Davis
Title-Subject: [Filtered] LKB type 7801B Knife Maker
Question: We have inherited an LKB type 7801B glass knife maker that doesn't seem to be working. As a rather general question, can anyone tell me whether it is possible and/or practical to have such an instrument fixed? Any help or suggestions would be welcome, and thanks in advance.
I received many useful answers and observations about Co calibration in Oxford ISIS 300 EDS system. I wish to thank everyone who answered me on this list and on private email, now I am able to do some considerations on the stability of our system and quality of our analyses Pietro
-- ********************************************* Pietro Arico' Dipartimento di Chimica e fisica della Terra ed Applicazioni alle Georisorse ed ai Rischi Naturali (CFTA) University of Palermo VIa Archirafi, 36 Palermo, 90123 - Italy email: p.arico-at-email.it Tel. +390916161574 (ext. 139) *********************************************
==============================Original Headers============================== 4, 18 -- From p.arico-at-email.it Thu Mar 30 00:33:53 2006 4, 18 -- Received: from www.unipa.it (www.unipa.it [147.163.1.5]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2U6Xqe4014656 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 30 Mar 2006 00:33:52 -0600 4, 18 -- Received: from [147.163.124.22] ([147.163.124.22]) 4, 18 -- by www.unipa.it (8.13.1/8.13.1) with ESMTP id k2U6UFnU019707 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 30 Mar 2006 08:30:15 +0200 4, 18 -- Message-ID: {442B7B38.5090909-at-email.it} 4, 18 -- Date: Thu, 30 Mar 2006 08:31:20 +0200 4, 18 -- From: "Pietro Arico'" {p.arico-at-email.it} 4, 18 -- User-Agent: Mozilla Thunderbird 1.5 (Windows/20051201) 4, 18 -- MIME-Version: 1.0 4, 18 -- To: MSA {Microscopy-at-msa.microscopy.com} 4, 18 -- Subject: EDS - Co calibration - thank you! 4, 18 -- Content-Type: text/plain; charset=ISO-8859-15; format=flowed 4, 18 -- Content-Transfer-Encoding: 7bit 4, 18 -- X-Virus-Scanned: ClamAV version 0.88, clamav-milter version 0.87 on www.unipa.it 4, 18 -- X-Virus-Status: Clean ==============================End of - Headers==============================
Hi, we are still using old LKB 7801A Knife Maker and it is still working well. It is a solid device.
Oldrich
Institute of Microbiology Acad. Sci. CR EM Lab Czech Republic
} Email: dgmorgan-at-ucdavis.edu } Name: David Morgan } } Organization: UC Davis } } Title-Subject: [Filtered] LKB type 7801B Knife Maker } } Question: We have inherited an LKB type 7801B glass knife maker that } doesn't seem to be working. As a rather general question, can anyone tell } me whether it is possible and/or practical to have such an instrument } fixed? Any help or suggestions would be welcome, and thanks in advance. }
==============================Original Headers============================== 7, 25 -- From benada-at-biomed.cas.cz Thu Mar 30 01:26:15 2006 7, 25 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2U7QE7Z024694 7, 25 -- for {microscopy-at-microscopy.com} ; Thu, 30 Mar 2006 01:26:14 -0600 7, 25 -- Received: from mail2.biomed.cas.cz (localhost [127.0.0.1]) 7, 25 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k2U7PxJx023232; 7, 25 -- Thu, 30 Mar 2006 09:25:59 +0200 (CEST) 7, 25 -- Received: from 147.231.44.104 7, 25 -- (SquirrelMail authenticated user benada) 7, 25 -- by mail2.biomed.cas.cz with HTTP; 7, 25 -- Thu, 30 Mar 2006 09:25:59 +0200 (CEST) 7, 25 -- Message-ID: {1260.147.231.44.104.1143703559.squirrel-at-mail2.biomed.cas.cz} 7, 25 -- In-Reply-To: {200603300009.k2U09Ynb023609-at-ns.microscopy.com} 7, 25 -- References: {200603300009.k2U09Ynb023609-at-ns.microscopy.com} 7, 25 -- Date: Thu, 30 Mar 2006 09:25:59 +0200 (CEST) 7, 25 -- Subject: Re: [Microscopy] viaWWW: LKB type 7801B Knife Maker 7, 25 -- From: =?iso-8859-2?Q?Old=F8ich_Benada?= {benada-at-biomed.cas.cz} 7, 25 -- To: dgmorgan-at-ucdavis.edu 7, 25 -- Cc: microscopy-at-microscopy.com 7, 25 -- User-Agent: SquirrelMail/1.4.6 7, 25 -- MIME-Version: 1.0 7, 25 -- Content-Type: text/plain;charset=iso-8859-2 7, 25 -- Content-Transfer-Encoding: 8bit 7, 25 -- X-Priority: 3 (Normal) 7, 25 -- Importance: Normal ==============================End of - Headers==============================
We have an (ancient) Hummer 1 coater in our lab that is in good working order, but the argon valve is showing signs of age. It has been replaced in the past, but we do not know of a good source for needle valves of this type. Does anyone have a good source for a replacement, or a salvaged needle valve in a drawer somewhere?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 3, 23 -- From hagglundk1-at-nku.edu Thu Mar 30 12:56:10 2006 3, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2UIuAol019833 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 30 Mar 2006 12:56:10 -0600 3, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 3, 23 -- Thu, 30 Mar 2006 13:56:10 -0500 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 3, 23 -- Content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="us-ascii" 3, 23 -- Subject: Replacement Argon Valve (Hummer 1) 3, 23 -- Date: Thu, 30 Mar 2006 13:56:09 -0500 3, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3586CB-at-mailfac1.hh.nku.edu} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: Replacement Argon Valve (Hummer 1) 3, 23 -- Thread-Index: AcZUK5tAXzEUCjwHSM+7k5PAG7XC1A== 3, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 3, 23 -- To: {microscopy-at-microscopy.com} 3, 23 -- X-OriginalArrivalTime: 30 Mar 2006 18:56:10.0165 (UTC) FILETIME=[9BD8EE50:01C6542B] 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2UIuAol019833 ==============================End of - Headers==============================
You might try swagelok: http://www.swagelok.com/search/find_products.aspx
Search for: metering valve
Regards, Woody White BWXT Services
-----Original Message----- X-from: hagglundk1-at-nku.edu [mailto:hagglundk1-at-nku.edu] Sent: Thursday, March 30, 2006 1:57 PM To: White, Woody N.
We have an (ancient) Hummer 1 coater in our lab that is in good working order, but the argon valve is showing signs of age. It has been replaced in the past, but we do not know of a good source for needle valves of this type. Does anyone have a good source for a replacement, or a salvaged needle valve in a drawer somewhere?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 3, 23 -- From hagglundk1-at-nku.edu Thu Mar 30 12:56:10 2006 3, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2UIuAol019833 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 30 Mar 2006 12:56:10 -0600 3, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.211); 3, 23 -- Thu, 30 Mar 2006 13:56:10 -0500 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 3, 23 -- Content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="us-ascii" 3, 23 -- Subject: Replacement Argon Valve (Hummer 1) 3, 23 -- Date: Thu, 30 Mar 2006 13:56:09 -0500 3, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F3586CB-at-mailfac1.hh.nku.edu} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: Replacement Argon Valve (Hummer 1) 3, 23 -- Thread-Index: AcZUK5tAXzEUCjwHSM+7k5PAG7XC1A== 3, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 3, 23 -- To: {microscopy-at-microscopy.com} 3, 23 -- X-OriginalArrivalTime: 30 Mar 2006 18:56:10.0165 (UTC) FILETIME=[9BD8EE50:01C6542B] 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2UIuAol019833 ==============================End of - Headers==============================
==============================Original Headers============================== 15, 27 -- From nwwhite-at-bwxt.com Thu Mar 30 13:23:22 2006 15, 27 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 15, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2UJNMvm029873 15, 27 -- for {microscopy-at-msa.microscopy.com} ; Thu, 30 Mar 2006 13:23:22 -0600 15, 27 -- Received: from ([131.184.13.224]) 15, 27 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.593521; 15, 27 -- Thu, 30 Mar 2006 14:23:11 -0500 15, 27 -- Received: from bwxslynpo01.BWXS.BWXTECH.NET ([131.184.13.4]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 15, 27 -- Thu, 30 Mar 2006 14:23:11 -0500 15, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 27 -- Content-class: urn:content-classes:message 15, 27 -- MIME-Version: 1.0 15, 27 -- Content-Type: text/plain; 15, 27 -- charset="US-ASCII" 15, 27 -- Subject: RE: [Microscopy] Replacement Argon Valve (Hummer 1) 15, 27 -- Date: Thu, 30 Mar 2006 14:23:11 -0500 15, 27 -- Message-ID: {70E4EA352EC987489773D55B0FF83E1C1DF9CB-at-bwxslynpo01.BWXS.BWXTECH.NET} 15, 27 -- X-MS-Has-Attach: 15, 27 -- X-MS-TNEF-Correlator: 15, 27 -- Thread-Topic: [Microscopy] Replacement Argon Valve (Hummer 1) 15, 27 -- Thread-Index: AcZUK8l3Kub/AeK/Q+Sz3HLhe16bdAAA1itA 15, 27 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 15, 27 -- To: {hagglundk1-at-nku.edu} , 15, 27 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 15, 27 -- X-OriginalArrivalTime: 30 Mar 2006 19:23:11.0738 (UTC) FILETIME=[626155A0:01C6542F] 15, 27 -- Content-Transfer-Encoding: 8bit 15, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2UJNMvm029873 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both ecd10-at-psu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Filtered] Postdoctoral Position
Question: POST-DOCTORAL POSITION in Transmission Electron Microscopy of Amorphous Thin Films for IR Sensing at The Pennsylvania State University
A postdoctoral position is available in the area of transmission electron microscopy beginning May 1, 2006. The research project focuses on understanding structure and chemistry of amorphous metal-oxides and semiconducting materials for IR sensor applications. Through a variety of electron imaging, spectroscopy and diffraction (e.g. fluctuation EM) techniques we aim to quantify structure and chemistry of amorphous thin films to establish processing/structure/property relationships for this class of materials. Furthermore, we aim to understand the microstructural and microchemical evolution under thermal and electrical stress. Most of the research will be conducted on a JEOL 2010F Field Emission TEM/STEM outfitted with an EDAX energy dispersive x-ray spectrometer (EDS), Gatan Enfina electron energy loss spectrometer (EELS), high-angle annular dark field STEM detector, and acquisition hardware and software for spectrum imaging. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
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Email: dm24-at-cornell.edu Name: David A. Muller
Organization: Cornell university
Title-Subject: [Filtered] Cornell Workshop & Summer School July 13-20 , 2006
Question: On the thirtieth anniversary of the first Cornell workshop on analytical electron microscopy, as well as John Silcox's 45th anniversary at Cornell, the Kavli Institute at Cornell will host a third summer school and workshop from July 13, to July 20, 2006. The conference website is http://www.research.cornell.edu/kic/events/em2006 and a poster is attached.
Electron microscopy has been at the forefront of nanoscale studies of materials, directly probing both physical and electronic properties. From the imaging and spectroscopy of individual dopant atoms and clusters buried inside a semiconductor host to the three-dimensional tomography of nanoparticles and biological structures, and the in-situ observations of nanomechanical deformations and electrodeposition, advances in instrumentation and algorithms have dramatically changed the field of electron microscopy. Early results in sub-angstrom resolution and millivolt spectroscopy are now being applied to materials problems, and international initiatives in aberration-corrected instruments should make such facilities available to the wider community.
The Summer School (July 13-15) will explore general theory of imaging, multislice and Bloch-wave simulations, and the theory of electron energy loss spectroscopy; computer labs are included. The workshop (July 16-20) will assess the present state of analytical electron microscopy and its impact on the physical and biological sciences, and identify the fundamental limits and the new science that next-generation technology should make possible.
Workshop invited speakers include:
Les Allen, University of Melbourne Phil Batson, IBM T.J. Watson Research Center Gianluigi Botton, McMaster University Nigel Browning, LLNL/UC Davis Christian Colliex, CNRS, UniversitČ Paris Sud John Cummings, University of Maryland Joachim Frank, HHMI, Wadsworth Center Archie Howie, University of Cambridge Ondrej Krivanek, NION Co. Richard Leapman, National Institutes of Health David Muller, Cornell University Harald Rose, Technical University of Darmstadt Frances Ross, IBM, T.J. Watson Research Center John Spence, Arizona State University Suzanne Stemmer, UC Santa Barbara Akira Tonomura, Hitachi Advanced Research Laboratory Maria Varela, Oak Ridge National Laboratory David Williams, Lehigh University Nestor Zaluzec, Argonne National Laboratory
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 eoptics-at-mcmaster.ca as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: eoptics-at-mcmaster.ca Name: Fred Pearson
Organization: McMaster University
Title-Subject: [Filtered] TEM-- Double Extraction Replicas for SuperAlloys
Question: I have a user who requires a procedure for looking at super alloy gamma(prime)phase using the double extraction replica method. An procedure from text or from an article would be appreciated. The method involved plastic replica and wax, dissolved in toluene, then carbon coating and shadow casting with Cr.
We are viewing bacteria plated out on filter membranes in the SEM and are having the standard problem of the biofilm/extracellular polysaccharide/mucus/slime around the organisms curdling up during specimen preparation. Normally it's not a huge problem, but this batch of bugs seems to be exceptionally slimy and the stuff is obscuring the structures we want to see.
My question is: are there any techniques for eliminating this substance before or during processing in order to see unBlemished Bugs. As usual, I'm searching for this information in all the usual places, but want to check to see if you all have any pet techniques.
Thanks! Enjoy the weekend.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 11, 23 -- From TindallR-at-missouri.edu Fri Mar 31 13:05:35 2006 11, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 11, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k2VJ5ZDo022885 11, 23 -- for {microscopy-at-microscopy.com} ; Fri, 31 Mar 2006 13:05:35 -0600 11, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 11, 23 -- Fri, 31 Mar 2006 13:05:34 -0600 11, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 11, 23 -- Content-class: urn:content-classes:message 11, 23 -- MIME-Version: 1.0 11, 23 -- Content-Type: text/plain; 11, 23 -- charset="us-ascii" 11, 23 -- Subject: SEM: Bacterial Biofilm Blues 11, 23 -- Date: Fri, 31 Mar 2006 13:05:33 -0600 11, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849E69-at-UM-EMAIL09.um.umsystem.edu} 11, 23 -- X-MS-Has-Attach: 11, 23 -- X-MS-TNEF-Correlator: 11, 23 -- Thread-Topic: SEM: Bacterial Biofilm Blues 11, 23 -- thread-index: AcZU9hZwLwb+4wqzTKq02m0f0Q5MmQ== 11, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 11, 23 -- To: {microscopy-at-microscopy.com} 11, 23 -- X-OriginalArrivalTime: 31 Mar 2006 19:05:34.0295 (UTC) FILETIME=[16821670:01C654F6] 11, 23 -- Content-Transfer-Encoding: 8bit 11, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k2VJ5ZDo022885 ==============================End of - Headers==============================
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
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Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
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:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
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:
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} } 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 (20Ĺ) 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
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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.
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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
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Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers Institute
Title-Subject: [Filtered] Leica EM-Stain
Question: Hello, I am interested in purchasing the Leica EM-Stain. Does anyone have any comments, positive and negative, that I should take into consideration before making such an investment? We will be using it on formvar coated slot grids. Thanks in advance, Rhonda Allen BA HT(ASCP)HTL, QIHC Stowers Institute for Medical Research Kansas City, Missouri 816-926-4346
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
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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 56C) at room temperature (about 18C) 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
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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
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==============================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
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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.
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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
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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
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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:
ď Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ď Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Leiden University Medical Center, Netherlands)
ď 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); Michael Rademacher (Univ. of Vermont)
ď Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ď Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ď 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==============================
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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
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==============================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==============================
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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.
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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==============================
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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
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==============================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.
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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
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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==============================
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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==============================
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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
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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
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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:
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--
********************************* 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:
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*********************************************** 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:
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==============================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
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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:
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} 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
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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
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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.
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==============================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==============================
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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...
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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==============================
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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.
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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==============================
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==============================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
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==============================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:
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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:
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********************************* 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
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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 ===
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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
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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
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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 _____________________________________________________________
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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 **********************************************
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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 } **********************************************
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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:
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Analytical Electron Microscopy at the Nanometer-Scale (June 12-15)
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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.
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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...
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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.
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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
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At 08:27 AM 4/21/2006, chakravarthi-at-ccmb.res.in wrote:
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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
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At 08:28 AM 4/21/2006, wickram-at-engr.colostate.edu wrote:
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==============================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
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==============================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:
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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: } } } ----------------------------------------------------------------------------
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} } 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:
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} } 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: } } } } } } } } } } ----------------------------------------------------------------------------
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} } } } } 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 ===
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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
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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
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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
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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
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-- __ 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"
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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
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... 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
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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==============================
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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.
A very good day to all of you. I would like to know if anyone can direct me to the companies that are selling ZnO Epitaxial Films. I have been searching around but with no results.
I thank you all in advance first!
Have a good day ahead!
Cheers, Yee Yan School of Materials Science and Engineering University of New South Wales
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==============================Original Headers============================== 7, 18 -- From one_twinklestar-at-yahoo.com.sg Mon May 1 04:01:07 2006 7, 18 -- Received: from web50010.mail.yahoo.com (web50010.mail.yahoo.com [206.190.38.25]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k41916Pu024269 7, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 1 May 2006 04:01:06 -0500 7, 18 -- Received: (qmail 91664 invoked by uid 60001); 1 May 2006 09:01:05 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com.sg; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=5X6mYpJ0adRKGCyi1HYiLfZgfD3n1UTOWYpfktlv7xh0XnsOsytsigvgbd+xaY4l9V3usDjmZ5b0VfJO7lD52mGmDdfilNAZcphfk4Nv5qlOFtelbjHojxXEm8wj+K16VM1ovWTUCpjFd/ahCOmx1JKwb56FxF9SR1P3rBp26DQ= ; 7, 18 -- Message-ID: {20060501090105.91662.qmail-at-web50010.mail.yahoo.com} 7, 18 -- Received: from [202.7.166.164] by web50010.mail.yahoo.com via HTTP; Mon, 01 May 2006 17:01:05 CST 7, 18 -- Date: Mon, 1 May 2006 17:01:05 +0800 (CST) 7, 18 -- From: Tay Yee Yan {one_twinklestar-at-yahoo.com.sg} 7, 18 -- Subject: Sales of Epitaxial Film-Enquires 7, 18 -- To: microscopy-at-msa.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==============================
This Question was submitted to Ask-A-Microscopist by (woad-at-iinet.net.au) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, April 30, 2006 at 21:45:31 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both woad-at-iinet.net.au as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: hi, I'm looking for 3 example images (moved out of phase by a 1/3 each) that have been recorded using structured illumination (say using a standing wave or otherwise projected) and the resultant merged hi-res version. I would like to apply the merging formula myself to the 3 images to see whether they turn out the way I hope them to i.e (that they are identical to a provided merged image). Hope you can help....
Sputter Films Inc (805-963-9651) sells a fluid lubricant (Torr Lube) that is advertised as being especially formulated for use on rotating and sliding seals, to have a vapor pressure below 10-6 Pa, and to provide a service life of up to 30 times that of most vacuum greases in such applications (See Vacuum Methods In Electron Microscopy , p. 460).
I have also found that the perfluorinated polyether diffusion pump fluids (ibid. p. 181) such as Fomblin Y-HVAC 25/9 and Krytox 1625 are very good lubricants for use on rotating and sliding vacuum seals, and might cost you a bit less than the Torr Lube. You can get these from most of the suppliers of EM products (Spi, Ladd, M. E. Taylor, etc.etc). -- 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 Mon May 1 14:28:37 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 k41JSbTb020718 2, 14 -- for {microscopy-at-microscopy.com} ; Mon, 1 May 2006 14:28:37 -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 (8.13.6/8.13.6) with ESMTP id k41JSaTT008861 2, 14 -- for {microscopy-at-microscopy.com} ; Mon, 1 May 2006 15:28:37 -0400 (EDT) 2, 14 -- Mime-Version: 1.0 2, 14 -- Message-Id: {p06210202c07c0e976fb0-at-[141.212.131.221]} 2, 14 -- Date: Mon, 1 May 2006 15:28:36 -0400 2, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 2, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 2, 14 -- Subject: [Microscopy] RE: Grease for ion millers 2, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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Email: kerverhe-at-msu.edu Name: Heather Kerver
Organization: Beaumont Hospital
Title-Subject: [Filtered] Diamond knives
Question: I was wondering what kind of glue is used to set the diamond knife. I am aware that it is a hydrophilic glue but what is the actual substance used?
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Email: carnahan-at-edison-labs.com Name: James Carnahan
Organization: Edison Analytical Laboratories, Inc.
Question: We had an unfortunate electrical accident that led to 120V going through part of the vacuum logic board from a failed turbo controller on an Amray 1700 Turbo. The console was off but the current blew the tops off of several of the logic chips and made a fuse out of one trace in the ground plane. We have some replacement chips and have found others on the net but are concerned about hidden damage. If anyone has an out-of-service 1700 Turbo or any Amray using the ES-7005-025 Vacuum logic board, we would be interested in purchasing the board.
Also, if anyone has experienced this problem we would like to hear their story.
Jim Carnahan Edison Analytical Labs (518) 393-2112
For the sake of SPI I just wanted to precise that the "high price and long delivery times" did not concern SPI products.
Stephane
--- "Garber, Charles A." {cgarber-at-2spi.com} wrote:
} But would you mind my asking you, were you } experiencing long delivery delays from SPI Supplies? } For Formvar coated grids, the delivery times are } usually pretty fast. I can't comment on what you } might perceive to be a high cost for purchased } filmed grids, most of our customers tell us that our } regular prices are lower than they could possibly } make them themselves.
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==============================Original Headers============================== 6, 19 -- From nizets2-at-yahoo.com Tue May 2 06:13:13 2006 6, 19 -- Received: from web37414.mail.mud.yahoo.com (web37414.mail.mud.yahoo.com [209.191.87.67]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k42BDDaX000481 6, 19 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 06:13:13 -0500 6, 19 -- Received: (qmail 7695 invoked by uid 60001); 2 May 2006 11:13:13 -0000 6, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 19 -- s=s1024; d=yahoo.com; 6, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 19 -- b=lRzT9FRlGWfwPLhUX3hDY8FRJljGPFlpmCuZqIULpOobEblfhouFaAU3dDt/S9Kngi5YREC11XcYwA+OxrsGNZceg09BDlyD85FyHaKues8ULyj3i4QKrOgkR93J+AJJkg4hEuf5d3gqXY9a0c27Ctd6J6L45nuJzZehm2rUGJ8= ; 6, 19 -- Message-ID: {20060502111313.7693.qmail-at-web37414.mail.mud.yahoo.com} 6, 19 -- Received: from [80.122.101.102] by web37414.mail.mud.yahoo.com via HTTP; Tue, 02 May 2006 04:13:13 PDT 6, 19 -- Date: Tue, 2 May 2006 04:13:13 -0700 (PDT) 6, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 19 -- Subject: Re: holes in the formvar film 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- In-Reply-To: {000c01c66a2d$1df584e0$b5750a0a-at-ibm1x23g2abfyg} 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: text/plain; charset=iso-8859-1 6, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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Organization: CNRS Laboratoire de Physique des Solides, Orsay
Title-Subject: [Filtered] engineer position in cryo-electron microscopy
Question: We are seeking an engineer in cryo-electron microscopy: The candidate will be in charge of a 200 kV LaB6 electron microscope equipped with a cryo-specimen holder and a post-column energy filter (GIF). He/she will conduct cryo-TEM experiments in biology as well as in physics of complex systems (polymers, colloids, amphiphiles, Ö), from specimen preparation to interpretation and image analysis. He/she must possess good abilities to design new and delicate experiments. Theoretical and practical background in transmission electron microscopy is required. Cryo-EM expertise will be appreciated. The candidate will integrate a multidisciplinary team "Structure and function of condensed DNA" located at the Solid State Physics Laboratory of the Paris-Sud University (Orsay, France). The position could suit either a physicist or a biologist with strong motivation for interdisciplinarity. Mastering of the English and French languages is required (the interview will be in French). Application deadline May 22.
Today's challenge is trying to image platinum nanoparticles in an agarose matrix, in order to see how they distribute themselves and to get a size distribution. TEM and/or SEM are possibilities. So far, my check of the literature finds tons of stuff on nanoparticles in electrophoresis gels, but none of it is relevant, since the particles are removed from the gels and put back into a liquid medium. The one article I found that is comparable to our problem used thin-sectioning and we have tried that.
We have also tried melting the agarose, dipping grids into the particle/agarose mix, then rinsing the grid in hot water to thin the gelatin out. We can get images in the TEM, but the results are inconsistent when repeating with the same sample. Also, there is the chance that the hot water and melting are re-arranging the particles.
We have tried thin sectioning the dehydrated agarose with particles, but finding the particles in a given thin section is a crap shoot with long odds. Also, we may be cutting through aggregations we want to see.
We have tried viewing carbon-coated dehydrated agarose with particles using backscattered electrons in our FESEM. This gives images with particles, but only those on or right at the surface are imaged clearly enough for good size data. Plus, we can't see far into the agarose for good distribution data.
We are going to try increasing the concentration of the particles to increase chances of getting them reliably in thin sections, and we will also try putting the melted mixture on cover slips in a thin layer and re-trying the BSE imaging after carbon coating. (The latter still has the potential problem of redistributing the particles, however.) We could also try doing large thick or semi-thin sections and viewing them in BSE imaging.
However, if someone out there has viewing nanos in agarose down to a fine art, we, as usual, would be delighted to hear about it. In the meantime, I will continue to search the databases.
Thanks to all!
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Immediately----Nano Takes a Little Longer! 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============================== 15, 23 -- From TindallR-at-missouri.edu Tue May 2 14:26:14 2006 15, 23 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) 15, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42JQE72005637 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 14:26:14 -0500 15, 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); 15, 23 -- Tue, 2 May 2006 14:26:11 -0500 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 23 -- Content-class: urn:content-classes:message 15, 23 -- MIME-Version: 1.0 15, 23 -- Content-Type: text/plain; 15, 23 -- charset="US-ASCII" 15, 23 -- Subject: TEM/SEM: Nanoparticles in agarose 15, 23 -- Date: Tue, 2 May 2006 14:26:11 -0500 15, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849F8F-at-UM-EMAIL09.um.umsystem.edu} 15, 23 -- X-MS-Has-Attach: 15, 23 -- X-MS-TNEF-Correlator: 15, 23 -- Thread-Topic: TEM/SEM: Nanoparticles in agarose 15, 23 -- thread-index: AcZuHkZUxNruEcQPRg2DlV4T+iVDcw== 15, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 15, 23 -- To: {microscopy-at-microscopy.com} 15, 23 -- X-OriginalArrivalTime: 02 May 2006 19:26:11.0424 (UTC) FILETIME=[451CFE00:01C66E1E] 15, 23 -- Content-Transfer-Encoding: 8bit 15, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k42JQE72005637 ==============================End of - Headers==============================
I have recently had a discussion with a colleague about the best protocol to follow when staining cells during a time course experiment. I don't think there is a single correct answer, however, would like to know current thinking on the following issue:
Live cells were treated with a compound and observed at various time points during a period of 48 hours. At each time point, cells were fixed and immunofluorescently stained for the protein of interest.
Is it a less artefactual procedure to fix cells at each time point and keep in a buffer until the end of 48 hours to stain them all at the same time or fix and stain at each sampling time point? To stain at the same time may would reduce staining differences, however, keeping cells in buffer for different times may induce changes in the protein.
I look forward to hearing your opinions. Thank you. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8, Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
==============================Original Headers============================== 7, 19 -- From trogadisj-at-smh.toronto.on.ca Tue May 2 14:59:26 2006 7, 19 -- Received: from smh.toronto.on.ca (mail.smh.toronto.on.ca [199.71.175.103]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42JxQwa015870 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 14:59:26 -0500 7, 19 -- Received: from ([199.71.171.15]) 7, 19 -- by beethoven.smh.toronto.on.ca with ESMTP id KP-GCT47.12851322; 7, 19 -- Tue, 02 May 2006 15:59:00 -0400 7, 19 -- Received: from SMTPOUT-MTA by beethoven.smh.toronto.on.ca 7, 19 -- with Novell_GroupWise; Tue, 02 May 2006 15:59:00 -0400 7, 19 -- Message-Id: {s45781c4.036-at-beethoven.smh.toronto.on.ca} 7, 19 -- X-Mailer: Novell GroupWise Internet Agent 6.5.2 7, 19 -- Date: Tue, 02 May 2006 15:58:48 -0400 7, 19 -- From: "Judy Trogadis" {TrogadisJ-at-smh.toronto.on.ca} 7, 19 -- To: {Microscopy-at-microscopy.com} 7, 19 -- Subject: time course experiment 7, 19 -- Mime-Version: 1.0 7, 19 -- Content-Type: text/plain; charset=US-ASCII 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- Content-Disposition: inline ==============================End of - Headers==============================
Judy, An end run around the problem is to start off the treatments at different times so they all end together and then fix and stain is all done at the same time. This doesn't answer your question but maybe a useful wrinkle?
Good luck, Tobias } } } Dear microscopists } } I have recently had a discussion with a colleague about the best } protocol to follow when staining cells during a time course experiment. } I don't think there is a single correct answer, however, would like to } know current thinking on the following issue: } } Live cells were treated with a compound and observed at various time } points during a period of 48 hours. At each time point, cells were fixed } and immunofluorescently stained for the protein of interest. } } Is it a less artefactual procedure to fix cells at each time point and } keep in a buffer until the end of 48 hours to stain them all at the same } time or fix and stain at each sampling time point? To stain at the same } time may would reduce staining differences, however, keeping cells in } buffer for different times may induce changes in the protein. } } I look forward to hearing your opinions. } Thank you. } Judy } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital, 7Queen } 30 Bond St. } Toronto, ON M5B 1W8, Canada } ph: 416-864-6060 x6337 } pager: 416-685-9219 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } ==============================Original Headers============================== } 7, 19 -- From trogadisj-at-smh.toronto.on.ca Tue May 2 14:59:26 2006 } 7, 19 -- Received: from smh.toronto.on.ca (mail.smh.toronto.on.ca } [199.71.175.103]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k42JxQwa015870 } 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 14:59:26 -0500 } 7, 19 -- Received: from ([199.71.171.15]) } 7, 19 -- by beethoven.smh.toronto.on.ca with ESMTP id } KP-GCT47.12851322; } 7, 19 -- Tue, 02 May 2006 15:59:00 -0400 } 7, 19 -- Received: from SMTPOUT-MTA by beethoven.smh.toronto.on.ca } 7, 19 -- with Novell_GroupWise; Tue, 02 May 2006 15:59:00 -0400 } 7, 19 -- Message-Id: {s45781c4.036-at-beethoven.smh.toronto.on.ca} } 7, 19 -- X-Mailer: Novell GroupWise Internet Agent 6.5.2 } 7, 19 -- Date: Tue, 02 May 2006 15:58:48 -0400 } 7, 19 -- From: "Judy Trogadis" {TrogadisJ-at-smh.toronto.on.ca} } 7, 19 -- To: {Microscopy-at-microscopy.com} } 7, 19 -- Subject: time course experiment } 7, 19 -- Mime-Version: 1.0 } 7, 19 -- Content-Type: text/plain; charset=US-ASCII } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- Content-Disposition: inline } ==============================End of - Headers==============================
There are some other things to consider. First off, a great deal of this debate will depend on what you are looking for and how it reacts with your fixative. If the cells are 'lightly fixed' there may be some reversal of fixation with prolonged buffer storage. Does that effect the staining? Tobias offered a good suggestion but there might be some chrono effects, cells fixed at different times of the day or night depending on your experimental design. I suggest avoiding all problems and debate by keeping all of the fixation, buffer wash times and staining times the same. Your staining proceedure should be sufficiently standardized so that it is not a variable, or is the least problematic of the potential variables. Finally, people looking for something to criticize in your proceedures will always find something 'wrong'.
Geoff
TrogadisJ-at-smh.toronto.on.ca wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 8, 35 -- From mcauliff-at-umdnj.edu Tue May 2 15:42:49 2006 8, 35 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 8, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42KgnJv003454 8, 35 -- for {microscopy-at-msa.microscopy.com} ; Tue, 2 May 2006 15:42:49 -0500 8, 35 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 8, 35 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 2E3001C3292 8, 35 -- for {microscopy-at-msa.microscopy.com} ; Tue, 2 May 2006 16:42:48 -0400 (EDT) 8, 35 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 8, 35 -- by zix01.umdnj.edu (Proprietary) with ESMTP id D37DCA7B42 8, 35 -- for {microscopy-at-msa.microscopy.com} ; Tue, 2 May 2006 16:42:46 -0400 (EDT) 8, 35 -- Received: from ([130.219.34.131]) 8, 35 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.12376868; 8, 35 -- Tue, 02 May 2006 16:42:19 -0400 8, 35 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 35 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 35 -- id {0IYN00M01NR1C0-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 35 -- for microscopy-at-msa.microscopy.com; Tue, 02 May 2006 16:42:19 -0400 (EDT) 8, 35 -- Received: from [127.0.0.1] ([10.138.2.123]) 8, 35 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 35 -- 2004)) with ESMTP id {0IYN008LMO5VGO-at-Polaris.umdnj.edu} ; Tue, 8, 35 -- 02 May 2006 16:41:56 -0400 (EDT) 8, 35 -- Date: Tue, 02 May 2006 16:42:52 -0400 8, 35 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 35 -- Subject: Re: [Microscopy] time course experiment 8, 35 -- In-reply-to: {200605022000.k42K0PwK017876-at-ns.microscopy.com} 8, 35 -- To: TrogadisJ-at-smh.toronto.on.ca, 8, 35 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 35 -- Message-id: {4457C44C.8030800-at-umdnj.edu} 8, 35 -- MIME-version: 1.0 8, 35 -- Content-type: text/plain; format=flowed; charset=us-ascii 8, 35 -- Content-transfer-encoding: 7BIT 8, 35 -- X-Accept-Language: en-us, en 8, 35 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 35 -- Gecko/20040804 Netscape/7.2 (ax) 8, 35 -- References: {200605022000.k42K0PwK017876-at-ns.microscopy.com} ==============================End of - Headers==============================
I don't know in detail what you want to observe, but isn't there a fluorescent tracker-molecule (such as a Lyso-tracker) available for your purpose? Another more ideal solution might be creating a, for that one protein GFP-positive cell-line?! Than you could make a continuous time-lapse without the need of fixation etc., all depending on your experiment's requests of course!
Anyway, if the fixation is strong enough, does the protein still show activity / are changes still induced? To my opinion and experience, if fixed strong enough and there are no/little changes, it should not matter whether the cells are stained immediately or a few hours later, especially when also stored cold. Best regards,
Sven Terclavers
-----Original Message----- X-from: TrogadisJ-at-smh.toronto.on.ca [mailto:TrogadisJ-at-smh.toronto.on.ca] Sent: dinsdag 2 mei 2006 22:02 To: sven.terclavers-at-med.kuleuven.be
Dear microscopists
I have recently had a discussion with a colleague about the best protocol to follow when staining cells during a time course experiment. I don't think there is a single correct answer, however, would like to know current thinking on the following issue:
Live cells were treated with a compound and observed at various time points during a period of 48 hours. At each time point, cells were fixed and immunofluorescently stained for the protein of interest.
Is it a less artefactual procedure to fix cells at each time point and keep in a buffer until the end of 48 hours to stain them all at the same time or fix and stain at each sampling time point? To stain at the same time may would reduce staining differences, however, keeping cells in buffer for different times may induce changes in the protein.
I look forward to hearing your opinions. Thank you. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8, Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
==============================Original Headers============================== 7, 19 -- From trogadisj-at-smh.toronto.on.ca Tue May 2 14:59:26 2006 7, 19 -- Received: from smh.toronto.on.ca (mail.smh.toronto.on.ca [199.71.175.103]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42JxQwa015870 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 14:59:26 -0500 7, 19 -- Received: from ([199.71.171.15]) 7, 19 -- by beethoven.smh.toronto.on.ca with ESMTP id KP-GCT47.12851322; 7, 19 -- Tue, 02 May 2006 15:59:00 -0400 7, 19 -- Received: from SMTPOUT-MTA by beethoven.smh.toronto.on.ca 7, 19 -- with Novell_GroupWise; Tue, 02 May 2006 15:59:00 -0400 7, 19 -- Message-Id: {s45781c4.036-at-beethoven.smh.toronto.on.ca} 7, 19 -- X-Mailer: Novell GroupWise Internet Agent 6.5.2 7, 19 -- Date: Tue, 02 May 2006 15:58:48 -0400 7, 19 -- From: "Judy Trogadis" {TrogadisJ-at-smh.toronto.on.ca} 7, 19 -- To: {Microscopy-at-microscopy.com} 7, 19 -- Subject: time course experiment 7, 19 -- Mime-Version: 1.0 7, 19 -- Content-Type: text/plain; charset=US-ASCII 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- Content-Disposition: inline ==============================End of - Headers==============================
==============================Original Headers============================== 17, 26 -- From Sven.Terclavers-at-med.kuleuven.be Tue May 2 16:36:50 2006 17, 26 -- Received: from hoboe2bl1.telenet-ops.be (hoboe2bl1.telenet-ops.be [195.130.137.73]) 17, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42LanKo014651 17, 26 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 16:36:49 -0500 17, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 17, 26 -- by hoboe2bl1.telenet-ops.be (Postfix) with SMTP 17, 26 -- id B58261243D6; Tue, 2 May 2006 23:36:48 +0200 (CEST) 17, 26 -- Received: from gwydion (d54C3B21A.access.telenet.be [84.195.178.26]) 17, 26 -- by hoboe2bl1.telenet-ops.be (Postfix) with ESMTP 17, 26 -- id 196E9124519; Tue, 2 May 2006 23:36:48 +0200 (CEST) 17, 26 -- Reply-To: {Sven.Terclavers-at-med.kuleuven.be} 17, 26 -- From: "Sven Terclavers" {Sven.Terclavers-at-med.kuleuven.be} 17, 26 -- To: {microscopy-at-microscopy.com} 17, 26 -- Cc: {TrogadisJ-at-smh.toronto.on.ca} 17, 26 -- Subject: RE: [Microscopy] time course experiment 17, 26 -- Date: Tue, 2 May 2006 23:36:46 +0200 17, 26 -- Organization: Home 17, 26 -- MIME-Version: 1.0 17, 26 -- Content-Type: text/plain; 17, 26 -- charset="US-ASCII" 17, 26 -- Content-Transfer-Encoding: 7bit 17, 26 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 17, 26 -- In-Reply-To: {200605022002.k42K26IW021687-at-ns.microscopy.com} 17, 26 -- Thread-Index: AcZuI0qIhRcgu4y2SSy9OSe9sNcMaQAC6iGA 17, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 17, 26 -- Message-Id: {20060502213648.196E9124519-at-hoboe2bl1.telenet-ops.be} ==============================End of - Headers==============================
This is a specimen prep question for everyone out there with EM expertise. We are having terrible success with Lead Citrate contrast staining. Principally, we suffer from precipitates showing up all over the specimen. On the advice of EM science technical support, we are double distilling our own water (they say Milli-Q is too pure and also is de-ionized which we don't want for EM). Then we make it CO2 free by autoclaving and capping directly upon removal from the autoclave. We do this the morning of reagent prep--so it doesn't sit in the bottle for longer than it takes to cool down before we begin making up the Reynolds.
We make the Reynolds Lead citrate according to the protocol listed in Ch 5 of Bozzola and Russell's Electron Microscopy (2nd edition) (mix 1.33g lead nitrate, 1.76g sodium citrate, and 30ml CO2 free double distilled water...shake vigorously for a few minutes and then again 5-6 times over the next 30 minutes). Ensure solution is milky white and free of particles. Add 8.0 ml commercially prepared, titrated 1.0 N NaOH--from EM science--solution turns clear. Adjust pH strictly to 12.0+/-0.1 unit. Bring volume to 50ml with CO2-free double distilled water. Stopper tightly with rubber stopper and parafilm until use later that day.
When we stain the grids with lead nitrate, we make sure to wash well before and after with CO2 free double distilled water in addition to surrounding the staining plate (Hiraoka kit) with NaOH pellets. In addition, we centrifuge the Reynolds at 5000xg for 8 minutes prior to use AND we 0.2um filter it into staining plate.
ANY advice or thoughts are welcome...what are we doing wrong?? What can we change about this protocol to ensure ppt free staining?
A thousand thanks in advance!
Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen-at-buckinstitute.org
==============================Original Headers============================== 9, 20 -- From dcrippen-at-buckinstitute.org Tue May 2 17:44:40 2006 9, 20 -- Received: from inverness.buckcenter.org (webmail.buckinstitute.org [64.84.58.24]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42MidAD025692 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 17:44:40 -0500 9, 20 -- Content-class: urn:content-classes:message 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; 9, 20 -- charset="iso-8859-1" 9, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 20 -- Subject: TEM--Lead Citrate--HELP!! 9, 20 -- Date: Tue, 2 May 2006 15:44:39 -0700 9, 20 -- Message-ID: {C7CAAAC7D2D14C409367E8D9026C1D78162E31-at-inverness.buckcenter.org} 9, 20 -- X-MS-Has-Attach: 9, 20 -- X-MS-TNEF-Correlator: 9, 20 -- Thread-Topic: TEM--Lead Citrate--HELP!! 9, 20 -- Thread-Index: AcZuOf68iZ+IkEf+SMS5wpYa7hbUNw== 9, 20 -- From: "Danielle Crippen" {dcrippen-at-buckinstitute.org} 9, 20 -- To: {microscopy-at-microscopy.com} 9, 20 -- Content-Transfer-Encoding: 8bit 9, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k42MidAD025692 ==============================End of - Headers==============================
Are you SURE it's lead citrate precip? There are many other sources of precipitates and "pepper", as I'm sure you're aware. What kind of sample are you preparing? What buffer is being used? Are you osmicating your samples?
We fought a pepper problem for over two years, before finally discovering that adding 2-mercaptoethanol to our buffer solved the problem.
Feel free to email an image, if you like.
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: dcrippen-at-buckinstitute.org [mailto:dcrippen-at-buckinstitute.org] Sent: Tuesday, May 02, 2006 5:47 PM To: Tindall, Randy D.
Dear TEM users,
This is a specimen prep question for everyone out there with EM expertise. We are having terrible success with Lead Citrate contrast staining. Principally, we suffer from precipitates showing up all over the specimen. On the advice of EM science technical support, we are double distilling our own water (they say Milli-Q is too pure and also is de-ionized which we don't want for EM). Then we make it CO2 free by autoclaving and capping directly upon removal from the autoclave. We do this the morning of reagent prep--so it doesn't sit in the bottle for longer than it takes to cool down before we begin making up the Reynolds.
We make the Reynolds Lead citrate according to the protocol listed in Ch 5 of Bozzola and Russell's Electron Microscopy (2nd edition) (mix 1.33g lead nitrate, 1.76g sodium citrate, and 30ml CO2 free double distilled water...shake vigorously for a few minutes and then again 5-6 times over the next 30 minutes). Ensure solution is milky white and free of particles. Add 8.0 ml commercially prepared, titrated 1.0 N NaOH--from EM science--solution turns clear. Adjust pH strictly to 12.0+/-0.1 unit. Bring volume to 50ml with CO2-free double distilled water. Stopper tightly with rubber stopper and parafilm until use later that day.
When we stain the grids with lead nitrate, we make sure to wash well before and after with CO2 free double distilled water in addition to surrounding the staining plate (Hiraoka kit) with NaOH pellets. In addition, we centrifuge the Reynolds at 5000xg for 8 minutes prior to use AND we 0.2um filter it into staining plate.
ANY advice or thoughts are welcome...what are we doing wrong?? What can we change about this protocol to ensure ppt free staining?
A thousand thanks in advance!
Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen-at-buckinstitute.org
==============================Original Headers============================== 9, 20 -- From dcrippen-at-buckinstitute.org Tue May 2 17:44:40 2006 9, 20 -- Received: from inverness.buckcenter.org (webmail.buckinstitute.org [64.84.58.24]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42MidAD025692 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 17:44:40 -0500 9, 20 -- Content-class: urn:content-classes:message 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; 9, 20 -- charset="iso-8859-1" 9, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 20 --
Danielle,
I haven't worked in biological electron microscopy for over 25 years. However, I vividly remember a colleague having a terrible time with precipitates from lead citrate staining. Turned out that the problem was his eye-sight. He was extremely near-sighted. To watch his work, his face was only several inches from the staining grid and water rinse. The source of the problem was CO2 from his breath. Another colleague happened to see his proximity to the stain and suggested the source of the problem. The precipitates disappeared once he isolated his exhaust from the process.
Food for thought.
Regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
dcrippen-at-bucki nstitute.org To gary.m.brown-at-exxonmobil.com 05/02/06 05:48 cc PM Subject [Microscopy] TEM--Lead Please respond Citrate--HELP!! to dcrippen-at-bucki nstitute.org
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Dear TEM users,
This is a specimen prep question for everyone out there with EM expertise. We are having terrible success with Lead Citrate contrast staining. Principally, we suffer from precipitates showing up all over the specimen. On the advice of EM science technical support, we are double distilling our own water (they say Milli-Q is too pure and also is de-ionized which we don't want for EM). Then we make it CO2 free by autoclaving and capping directly upon removal from the autoclave. We do this the morning of reagent prep--so it doesn't sit in the bottle for longer than it takes to cool down before we begin making up the Reynolds.
We make the Reynolds Lead citrate according to the protocol listed in Ch 5 of Bozzola and Russell's Electron Microscopy (2nd edition) (mix 1.33g lead nitrate, 1.76g sodium citrate, and 30ml CO2 free double distilled water...shake vigorously for a few minutes and then again 5-6 times over the next 30 minutes). Ensure solution is milky white and free of particles. Add 8.0 ml commercially prepared, titrated 1.0 N NaOH--from EM science--solution turns clear. Adjust pH strictly to 12.0+/-0.1 unit. Bring volume to 50ml with CO2-free double distilled water. Stopper tightly with rubber stopper and parafilm until use later that day.
When we stain the grids with lead nitrate, we make sure to wash well before and after with CO2 free double distilled water in addition to surrounding the staining plate (Hiraoka kit) with NaOH pellets. In addition, we centrifuge the Reynolds at 5000xg for 8 minutes prior to use AND we 0.2um filter it into staining plate.
ANY advice or thoughts are welcome...what are we doing wrong?? What can we change about this protocol to ensure ppt free staining?
A thousand thanks in advance!
Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen-at-buckinstitute.org
==============================Original Headers============================== 9, 20 -- From dcrippen-at-buckinstitute.org Tue May 2 17:44:40 2006 9, 20 -- Received: from inverness.buckcenter.org (webmail.buckinstitute.org [64.84.58.24]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42MidAD025692 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 17:44:40 -0500 9, 20 -- Content-class: urn:content-classes:message 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; 9, 20 -- charset="iso-8859-1" 9, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 20 -- Subject: TEM--Lead Citrate--HELP!! 9, 20 -- Date: Tue, 2 May 2006 15:44:39 -0700 9, 20 -- Message-ID: {C7CAAAC7D2D14C409367E8D9026C1D78162E31-at-inverness.buckcenter.org} 9, 20 -- X-MS-Has-Attach: 9, 20 -- X-MS-TNEF-Correlator: 9, 20 -- Thread-Topic: TEM--Lead Citrate--HELP!! 9, 20 -- Thread-Index: AcZuOf68iZ+IkEf+SMS5wpYa7hbUNw== 9, 20 -- From: "Danielle Crippen" {dcrippen-at-buckinstitute.org} 9, 20 -- To: {microscopy-at-microscopy.com} 9, 20 -- Content-Transfer-Encoding: 8bit 9, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k42MidAD025692 ==============================End of - Headers==============================
==============================Original Headers============================== 28, 19 -- From gary.m.brown-at-exxonmobil.com Tue May 2 18:10:15 2006 28, 19 -- Received: from hoespc01.exxonmobil.com (hoespc01.exxonmobil.com [192.67.48.38]) 28, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42NAFLn013101 28, 19 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 18:10:15 -0500 28, 19 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 28, 19 -- by hoespc01.exxonmobil.com (Switch-3.1.7/Switch-3.1.7) with ESMTP id k42N9roU006716; 28, 19 -- Tue, 2 May 2006 18:09:54 -0500 (CDT) 28, 19 -- In-Reply-To: {200605022248.k42Mm0ti029416-at-ns.microscopy.com} 28, 19 -- Subject: Re: [Microscopy] TEM--Lead Citrate--HELP!! 28, 19 -- Importance: 28, 19 -- To: dcrippen-at-buckinstitute.org, microscopy-at-microscopy.com 28, 19 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 28, 19 -- Message-ID: {OF1CB0C5B8.3466EB9F-ON86257162.007E1D3A-86257162.007F44B0-at-exxonmobil.com} 28, 19 -- From: gary.m.brown-at-exxonmobil.com 28, 19 -- Date: Tue, 2 May 2006 18:10:07 -0500 28, 19 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652FP1HF193|March 28, 19 -- 02, 2006) at 05/02/2006 06:10:12 PM 28, 19 -- MIME-Version: 1.0 28, 19 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
On May 2, 2006, at 12:26 PM, TindallR-at-missouri.edu wrote:
} Today's challenge is trying to image platinum nanoparticles in an } agarose matrix, in order to see how they distribute themselves and to } get a size distribution. TEM and/or SEM are possibilities. So far, my } check of the literature finds tons of stuff on nanoparticles in } electrophoresis gels, but none of it is relevant, since the particles } are removed from the gels and put back into a liquid medium. The one } article I found that is comparable to our problem used thin-sectioning } and we have tried that. } } We have also tried melting the agarose, dipping grids into the } particle/agarose mix, then rinsing the grid in hot water to thin the } gelatin out. We can get images in the TEM, but the results are } inconsistent when repeating with the same sample. Also, there is the } chance that the hot water and melting are re-arranging the particles. } } We have tried thin sectioning the dehydrated agarose with particles, } but } finding the particles in a given thin section is a crap shoot with long } odds. Also, we may be cutting through aggregations we want to see. } } We have tried viewing carbon-coated dehydrated agarose with particles } using backscattered electrons in our FESEM. This gives images with } particles, but only those on or right at the surface are imaged clearly } enough for good size data. Plus, we can't see far into the agarose for } good distribution data. } } We are going to try increasing the concentration of the particles to } increase chances of getting them reliably in thin sections, and we will } also try putting the melted mixture on cover slips in a thin layer and } re-trying the BSE imaging after carbon coating. (The latter still has } the potential problem of redistributing the particles, however.) We } could also try doing large thick or semi-thin sections and viewing them } in BSE imaging. } } However, if someone out there has viewing nanos in agarose down to a } fine art, we, as usual, would be delighted to hear about it. In the } meantime, I will continue to search the databases. } Dear Randy, Thick or semi-thick sections in TEM would be my choice--probably since I have a 300 kV TEM. If you can get to a high-pressure freezer, I would suggest using that to prepare your specimens, followed by freeze-substitution and resin embedding. Assuming that you do not need to image the strands of agarose, just section the embedded specimen and observe. 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 May 2 18:22:21 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 k42NML6K022993 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 2 May 2006 18:22:21 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP 4, 22 -- id 0412A361B2; Tue, 2 May 2006 16:22:21 -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 F421210AB45; Tue, 2 May 2006 16:22:19 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200605021926.k42JQQx1005807-at-ns.microscopy.com} 4, 22 -- References: {200605021926.k42JQQx1005807-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {550efc1d94e877099710308bf41bdfde-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] TEM/SEM: Nanoparticles in agarose 4, 22 -- Date: Tue, 2 May 2006 16:32:36 -0700 4, 22 -- To: "Randy D.Tindall" {TindallR-at-missouri.edu} , microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.623) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Reaching into my past......We used special analytical grade NaOH, since it seemed to us that the NaOH was picking up carbonate from the air. The analytical grade stuff came in a sealed glass vial, and made quite a difference.
Joel
Date sent: Tue, 2 May 2006 17:45:41 -0500 To: jbs-at-temple.edu X-from: dcrippen-at-buckinstitute.org Send reply to: dcrippen-at-buckinstitute.org
This may be shear luck, but I've never had trouble with precipitate (other things yes, but ppte no) - I keep my lead citrate (made up with ordinary distilled water, not specially CO2 free) in a volumetric flask (50ml), which sits in the same place month after month and is never moved. I don't use the stain for 24 hours after it's prepared, but then just carefully take off what's needed from close to the surface using a glass pipette. I wipe the end of the pipette with a tissue before dispensing the stain and then discard the first drop. I put the drops onto Parafilm in a covered glass petri dish. No need for NaOH pellets. Finally wash the grids for 5 seconds in a gentle stream of water from a wash bottle. I probably shouldn't admit this but I've had a bottle of stain last over 2 years (the surface of the bottle becomes cloudy with ppte) and still produce perfect results.
Cheers,
Diana
On 3 May 2006, at 8:47 AM, dcrippen-at-buckinstitute.org wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Dear TEM users, } } This is a specimen prep question for everyone out there with EM } expertise. We are having terrible success with Lead Citrate } contrast staining. Principally, we suffer from precipitates } showing up all over the specimen. On the advice of EM science } technical support, we are double distilling our own water (they say } Milli-Q is too pure and also is de-ionized which we don't want for } EM). Then we make it CO2 free by autoclaving and capping directly } upon removal from the autoclave. We do this the morning of reagent } prep--so it doesn't sit in the bottle for longer than it takes to } cool down before we begin making up the Reynolds. } } We make the Reynolds Lead citrate according to the protocol listed } in Ch 5 of Bozzola and Russell's Electron Microscopy (2nd edition) } (mix 1.33g lead nitrate, 1.76g sodium citrate, and 30ml CO2 free } double distilled water...shake vigorously for a few minutes and } then again 5-6 times over the next 30 minutes). Ensure solution is } milky white and free of particles. Add 8.0 ml commercially } prepared, titrated 1.0 N NaOH--from EM science--solution turns } clear. Adjust pH strictly to 12.0+/-0.1 unit. Bring volume to } 50ml with CO2-free double distilled water. Stopper tightly with } rubber stopper and parafilm until use later that day. } } When we stain the grids with lead nitrate, we make sure to wash } well before and after with CO2 free double distilled water in } addition to surrounding the staining plate (Hiraoka kit) with NaOH } pellets. In addition, we centrifuge the Reynolds at 5000xg for 8 } minutes prior to use AND we 0.2um filter it into staining plate. } } ANY advice or thoughts are welcome...what are we doing wrong?? } What can we change about this protocol to ensure ppt free staining? } } A thousand thanks in advance! } } Danielle Crippen } Morphology and Imaging Core Manager } Buck Institute for Age Research } 8001 Redwood Blvd. } Novato, CA 94945 } 415-209-2046 } dcrippen-at-buckinstitute.org } } } } ==============================Original } Headers============================== } 9, 20 -- From dcrippen-at-buckinstitute.org Tue May 2 17:44:40 2006 } 9, 20 -- Received: from inverness.buckcenter.org } (webmail.buckinstitute.org [64.84.58.24]) } 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k42MidAD025692 } 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 17:44:40 } -0500 } 9, 20 -- Content-class: urn:content-classes:message } 9, 20 -- MIME-Version: 1.0 } 9, 20 -- Content-Type: text/plain; } 9, 20 -- charset="iso-8859-1" } 9, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 9, 20 -- Subject: TEM--Lead Citrate--HELP!! } 9, 20 -- Date: Tue, 2 May 2006 15:44:39 -0700 } 9, 20 -- Message-ID: } {C7CAAAC7D2D14C409367E8D9026C1D78162E31-at-inverness.buckcenter.org} } 9, 20 -- X-MS-Has-Attach: } 9, 20 -- X-MS-TNEF-Correlator: } 9, 20 -- Thread-Topic: TEM--Lead Citrate--HELP!! } 9, 20 -- Thread-Index: AcZuOf68iZ+IkEf+SMS5wpYa7hbUNw== } 9, 20 -- From: "Danielle Crippen" {dcrippen-at-buckinstitute.org} } 9, 20 -- To: {microscopy-at-microscopy.com} } 9, 20 -- Content-Transfer-Encoding: 8bit } 9, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k42MidAD025692 } ==============================End of - } Headers==============================
==============================Original Headers============================== 7, 19 -- From dianavd-at-eye.usyd.edu.au Tue May 2 20:37:34 2006 7, 19 -- Received: from galen.med.usyd.edu.au (nugalen.med.usyd.edu.au [129.78.36.39]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k431bXgq011889 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 20:37:33 -0500 7, 19 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) 7, 19 -- by galen.med.usyd.edu.au with esmtp (Exim 4.43) 7, 19 -- id 1Fb6Ix-0001Sc-26; Wed, 03 May 2006 11:37:27 +1000 7, 19 -- Mime-Version: 1.0 (Apple Message framework v749.3) 7, 19 -- In-Reply-To: {200605022247.k42MlH5w028230-at-ns.microscopy.com} 7, 19 -- References: {200605022247.k42MlH5w028230-at-ns.microscopy.com} 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 19 -- Message-Id: {04E1FDBE-5BAF-4337-B5E4-2C18ED258184-at-eye.usyd.edu.au} 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} 7, 19 -- Subject: Re: [Microscopy] TEM--Lead Citrate--HELP!! 7, 19 -- Date: Wed, 3 May 2006 11:37:25 +1000 7, 19 -- To: Microscopy {microscopy-at-microscopy.com} , dcrippen-at-buckinstitute.org 7, 19 -- X-Mailer: Apple Mail (2.749.3) 7, 19 -- X-Spam-Score: -3.7 (---) ==============================End of - Headers==============================
We keep lead citrate prepared by reynold's method for months in a volumetric flask, Use Whatman 1 to filter and make drops on a parafilm in a petridish containing NaOH pellets. After staining for 1-3 min, wash the grids in water, then water with about 0.01% NaOH and again water. Never had any precipitates.
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear TEM users, } } This is a specimen prep question for everyone out } there with EM expertise. We are having terrible } success with Lead Citrate contrast staining. } Principally, we suffer from precipitates showing up } all over the specimen. On the advice of EM science } technical support, we are double distilling our own } water (they say Milli-Q is too pure and also is } de-ionized which we don't want for EM). Then we } make it CO2 free by autoclaving and capping directly } upon removal from the autoclave. We do this the } morning of reagent prep--so it doesn't sit in the } bottle for longer than it takes to cool down before } we begin making up the Reynolds. } } We make the Reynolds Lead citrate according to the } protocol listed in Ch 5 of Bozzola and Russell's } Electron Microscopy (2nd edition) (mix 1.33g lead } nitrate, 1.76g sodium citrate, and 30ml CO2 free } double distilled water...shake vigorously for a few } minutes and then again 5-6 times over the next 30 } minutes). Ensure solution is milky white and free } of particles. Add 8.0 ml commercially prepared, } titrated 1.0 N NaOH--from EM science--solution turns } clear. Adjust pH strictly to 12.0+/-0.1 unit. } Bring volume to 50ml with CO2-free double distilled } water. Stopper tightly with rubber stopper and } parafilm until use later that day. } } When we stain the grids with lead nitrate, we make } sure to wash well before and after with CO2 free } double distilled water in addition to surrounding } the staining plate (Hiraoka kit) with NaOH pellets. } In addition, we centrifuge the Reynolds at 5000xg } for 8 minutes prior to use AND we 0.2um filter it } into staining plate. } } ANY advice or thoughts are welcome...what are we } doing wrong?? What can we change about this } protocol to ensure ppt free staining? } } A thousand thanks in advance! } } Danielle Crippen } Morphology and Imaging Core Manager } Buck Institute for Age Research } 8001 Redwood Blvd. } Novato, CA 94945 } 415-209-2046 } dcrippen-at-buckinstitute.org } } } } ==============================Original } Headers============================== } 9, 20 -- From dcrippen-at-buckinstitute.org Tue May 2 } 17:44:40 2006 } 9, 20 -- Received: from inverness.buckcenter.org } (webmail.buckinstitute.org [64.84.58.24]) } 9, 20 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k42MidAD025692 } 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 } May 2006 17:44:40 -0500 } 9, 20 -- Content-class: urn:content-classes:message } 9, 20 -- MIME-Version: 1.0 } 9, 20 -- Content-Type: text/plain; } 9, 20 -- charset="iso-8859-1" } 9, 20 -- X-MimeOLE: Produced By Microsoft Exchange } V6.5 } 9, 20 -- Subject: TEM--Lead Citrate--HELP!! } 9, 20 -- Date: Tue, 2 May 2006 15:44:39 -0700 } 9, 20 -- Message-ID: } {C7CAAAC7D2D14C409367E8D9026C1D78162E31-at-inverness.buckcenter.org} } 9, 20 -- X-MS-Has-Attach: } 9, 20 -- X-MS-TNEF-Correlator: } 9, 20 -- Thread-Topic: TEM--Lead Citrate--HELP!! } 9, 20 -- Thread-Index: } AcZuOf68iZ+IkEf+SMS5wpYa7hbUNw== } 9, 20 -- From: "Danielle Crippen" } {dcrippen-at-buckinstitute.org} } 9, 20 -- To: {microscopy-at-microscopy.com} } 9, 20 -- Content-Transfer-Encoding: 8bit } 9, 20 -- X-MIME-Autoconverted: from quoted-printable } to 8bit by ns.microscopy.com id k42MidAD025692 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 6, 20 -- From shashis_99-at-yahoo.com Tue May 2 23:10:44 2006 6, 20 -- Received: from web54613.mail.yahoo.com (web54613.mail.yahoo.com [206.190.49.183]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k434AhDg024111 6, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 23:10:44 -0500 6, 20 -- Received: (qmail 76169 invoked by uid 60001); 3 May 2006 04:10:42 -0000 6, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 20 -- s=s1024; d=yahoo.com; 6, 20 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 20 -- b=ptQ7cT0W7aXsSDn7d2ujxNFCzmYsX6OWtiAAqbMZ/lfsNfYQNavioZdPDA2ZFfZH2w6s0IGPQFg/+g7kAbfXmKCNf1sFjy01pjBnf5VypfnA6HFkr3O2WiHAMjSKPo9oZ+H/s7uzILZehFv4AK5MOshoI/Tuu8UhcnIIIJs9lp8= ; 6, 20 -- Message-ID: {20060503041042.76167.qmail-at-web54613.mail.yahoo.com} 6, 20 -- Received: from [203.200.217.180] by web54613.mail.yahoo.com via HTTP; Tue, 02 May 2006 21:10:42 PDT 6, 20 -- Date: Tue, 2 May 2006 21:10:42 -0700 (PDT) 6, 20 -- From: shashi singh {shashis_99-at-yahoo.com} 6, 20 -- Subject: Re: [Microscopy] TEM--Lead Citrate--HELP!! 6, 20 -- To: dcrippen-at-buckinstitute.org, 6, 20 -- "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} 6, 20 -- In-Reply-To: {200605022247.k42MlX9q028672-at-ns.microscopy.com} 6, 20 -- MIME-Version: 1.0 6, 20 -- Content-Type: text/plain; charset=iso-8859-1 6, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Doesn't anybody else use or recommend Sato's lead stain as a more stable replacement for Reynolds Pb citrate? We've used it since the 1970s. 1968 Sato, T.: J. Electron Microsc. 17:158, 1968. 1968 Sato and others: Proc. XIth Int. Cong. on Electron Microscopy. Kyoto. 1986, pp. 2181-2182. [
-mike reedy-
At 5:49 PM -0500 5/2/06, dcrippen-at-buckinstitute.org wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
since it was not an issue in the replies so far, I would like to ask wether your double distilling apparatus overall is made of quartz glass or does have a destillate } container { bin made from metal (e. g. copper).
I only would like to add this since we had - several years ago - a problem when our destillation apparatus was out of function and on repair for some month and we used } bidistilled { water obtained from our hospital pharmacy. We had a lot of precipitation problems then, which ended not before we changed to the ddH2O from the repaired quartz-glass still used formerly.
When checking the quality of the "pharmacy"-water later on it turned out to contain a high amount of copper-ions (storage bin was made from copper sheets), which IMO perhaps might have had a detrimental precipitating action on the lead-staining performance.
By the way: we use Lead Citrate according to Venable&Coggeshall (1965), store solutions in "ultraclean" snap cap-glass-vials [and also ultraclean plastic snap cap!] which are used only for that purpose, that means we take care of any traces of cleaning substances by washing /cleaning also with chrome-sulfuric acid or a modern substitute but take care by ourselves (not a washing machine) to get rid of any resting traces of substances by vigorously washing several times with bidistilled hot water and a final step with ultrapure water (UHQ). We found also that intermittent air drying of glass vial/bottle creates probably otherwise insoluble incrustations, so we always keep the stuff in wet condition until the final step of cleaning.
Another point we found is that "freshly" made lead citrate solution (Venable&Coggeshall) -if used the same day - will be "more aggressive/more reactive", that means, we decrease staining times (say 30 sec when freshly prepared instead of 2-3 min -at-room temperature, e.g. after one week storage in the dark).
Avoiding or at least some sort of control for the CO2-reaction is obligatory in our lab (NaOH-pellets in a petridish filled with dental wax, the latter always being melted and } flamed/singed { after a staining cycle, but perhaps use of virgin, clean? { {Parafilm} } sheets is the better choice) knowing that some/severely disturbing precipitation nuclei also could be present in previousely uncleaned, and therefore } oily { injection needles, syringes, (plastic) tips, rubber stoppers (especially if always one and the same is used) as well as the surface areas where you are staining/handling your grids.
In general our experience is / was: the more steps you are introducing in your schedule to reduce an anticipated precipitate (or to inhibit the formation of such one) the more you (likely) will initiate precipitation due to unexpected particle impurities.
All best wishes for an excellent result of your next staining series,
Wolfgang Muss Salzburg Austria ---------- Von: dcrippen-at-buckinstitute.org[SMTP:dcrippen-at-buckinstitute.org] Antwort an: dcrippen-at-buckinstitute.org Gesendet: Mittwoch, 03. Mai 2006 00:50 An: W.Muss-at-salk.at Betreff: [Microscopy] TEM--Lead Citrate--HELP!!
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Dear TEM users,
This is a specimen prep question for everyone out there with EM expertise. We are having terrible success with Lead Citrate contrast staining. Principally, we suffer from precipitates showing up all over the specimen. On the advice of EM science technical support, we are double distilling our own water (they say Milli-Q is too pure and also is de-ionized which we don't want for EM). Then we make it CO2 free by autoclaving and capping directly upon removal from the autoclave. We do this the morning of reagent prep--so it doesn't sit in the bottle for longer than it takes to cool down before we begin making up the Reynolds.
We make the Reynolds Lead citrate according to the protocol listed in Ch 5 of Bozzola and Russell's Electron Microscopy (2nd edition) (mix 1.33g lead nitrate, 1.76g sodium citrate, and 30ml CO2 free double distilled water...shake vigorously for a few minutes and then again 5-6 times over the next 30 minutes). Ensure solution is milky white and free of particles. Add 8.0 ml commercially prepared, titrated 1.0 N NaOH--from EM science--solution turns clear. Adjust pH strictly to 12.0+/-0.1 unit. Bring volume to 50ml with CO2-free double distilled water. Stopper tightly with rubber stopper and parafilm until use later that day.
When we stain the grids with lead nitrate, we make sure to wash well before and after with CO2 free double distilled water in addition to surrounding the staining plate (Hiraoka kit) with NaOH pellets. In addition, we centrifuge the Reynolds at 5000xg for 8 minutes prior to use AND we 0.2um filter it into staining plate.
ANY advice or thoughts are welcome...what are we doing wrong?? What can we change about this protocol to ensure ppt free staining?
A thousand thanks in advance!
Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen-at-buckinstitute.org
==============================Original Headers============================== 9, 20 -- From dcrippen-at-buckinstitute.org Tue May 2 17:44:40 2006 9, 20 -- Received: from inverness.buckcenter.org (webmail.buckinstitute.org [64.84.58.24]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k42MidAD025692 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 2 May 2006 17:44:40 -0500 9, 20 -- Content-class: urn:content-classes:message 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; 9, 20 -- charset="iso-8859-1" 9, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 20 -- Subject: TEM--Lead Citrate--HELP!! 9, 20 -- Date: Tue, 2 May 2006 15:44:39 -0700 9, 20 -- Message-ID: {C7CAAAC7D2D14C409367E8D9026C1D78162E31-at-inverness.buckcenter.org} 9, 20 -- X-MS-Has-Attach: 9, 20 -- X-MS-TNEF-Correlator: 9, 20 -- Thread-Topic: TEM--Lead Citrate--HELP!! 9, 20 -- Thread-Index: AcZuOf68iZ+IkEf+SMS5wpYa7hbUNw== 9, 20 -- From: "Danielle Crippen" {dcrippen-at-buckinstitute.org} 9, 20 -- To: {microscopy-at-microscopy.com} 9, 20 -- Content-Transfer-Encoding: 8bit 9, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k42MidAD025692 ==============================End of - Headers==============================
==============================Original Headers============================== 24, 28 -- From W.Muss-at-salk.at Wed May 3 03:00:12 2006 24, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 24, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4380Bmm014853 24, 28 -- for {microscopy-at-microscopy.com} ; Wed, 3 May 2006 03:00:11 -0500 24, 28 -- Received: from localhost (localhost [127.0.0.1]) 24, 28 -- by hermes.lks.at (Postfix) with ESMTP id 6518A5A901F; 24, 28 -- Wed, 3 May 2006 10:00:04 +0200 (CEST) 24, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 24, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 24, 28 -- with ESMTP id 71716-05; Wed, 3 May 2006 10:00:04 +0200 (CEST) 24, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 24, 28 -- by hermes.lks.at (Postfix) with SMTP id 0FD325A900A; 24, 28 -- Wed, 3 May 2006 10:00:04 +0200 (CEST) 24, 28 -- Received: by localhost with Microsoft MAPI; Wed, 3 May 2006 10:00:03 +0200 24, 28 -- Message-ID: {01C66E98.5941B2C0.W.Muss-at-salk.at} 24, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 24, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 24, 28 -- To: "'dcrippen-at-buckinstitute.org'" {dcrippen-at-buckinstitute.org} , 24, 28 -- "'microscopy-at-microscopy.com'" {microscopy-at-microscopy.com} 24, 28 -- Subject: [Microscopy] Re: TEM--Lead Citrate--HELP!! 24, 28 -- Date: Wed, 3 May 2006 10:00:01 +0200 24, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 24, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 24, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 24, 28 -- MIME-Version: 1.0 24, 28 -- Content-Type: text/plain; charset="us-ascii" 24, 28 -- Content-Transfer-Encoding: 7bit 24, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Hi folks, Strictly this isn't a microscopy question but I think I have a fair chance of catching someone who might be able to help!
I was looking through our boxes of 'historical' samples (there has been a materials analysis lab here for more than 50 years), and I have examples of 1", 2", 3", 4", 6" and 8" Si wafers. Since we stopped getting Si samples to analyse about 5 years ago I don't have any 12" (or 300mm, I should say) wafers. Is there anyone out there willing to swap a 12" wafer for a 1" one? I know it doesn't seem like good value for the amount of material but I hope that would be more than compensated for by the historical interest. Or I could swap odd bits of Ge, GaAs, InP, sapphire, etc. etc. if Si isn't your bag. Perhaps I should start up a little 'Caswell museum'..
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==============================Original Headers============================== 9, 31 -- From richard.beanland-at-bookham.com Wed May 3 05:08:50 2006 9, 31 -- Received: from mail72.messagelabs.com (mail72.messagelabs.com [193.109.255.147]) 9, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k43A8oWW029818 9, 31 -- for {microscopy-at-microscopy.com} ; Wed, 3 May 2006 05:08:50 -0500 9, 31 -- X-VirusChecked: Checked 9, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 9, 31 -- X-Msg-Ref: server-14.tower-72.messagelabs.com!1146650928!34334684!1 9, 31 -- X-StarScan-Version: 5.5.9.1; banners=bookham.com,-,- 9, 31 -- X-Originating-IP: [213.249.209.179] 9, 31 -- Received: (qmail 6827 invoked from network); 3 May 2006 10:08:48 -0000 9, 31 -- Received: from unknown (HELO cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 9, 31 -- by server-14.tower-72.messagelabs.com with SMTP; 3 May 2006 10:08:48 -0000 9, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-01.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.211); 9, 31 -- Wed, 3 May 2006 11:08:48 +0100 9, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 9, 31 -- Content-class: urn:content-classes:message 9, 31 -- MIME-Version: 1.0 9, 31 -- Content-Type: text/plain; 9, 31 -- charset="us-ascii" 9, 31 -- Subject: Si wafers 9, 31 -- Date: Wed, 3 May 2006 11:08:47 +0100 9, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E083603-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 9, 31 -- X-MS-Has-Attach: 9, 31 -- X-MS-TNEF-Correlator: 9, 31 -- Thread-Topic: Si wafers 9, 31 -- Thread-Index: AcZumZF7SpnmhpHcRrOQHPRrNC+AIA== 9, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 9, 31 -- To: {microscopy-at-microscopy.com} 9, 31 -- X-OriginalArrivalTime: 03 May 2006 10:08:48.0562 (UTC) FILETIME=[9206F920:01C66E99] 9, 31 -- Content-Transfer-Encoding: 8bit 9, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k43A8oWW029818 ==============================End of - Headers==============================
} This may be shear luck, but I've never had trouble with } precipitate (other things yes, but ppte no) - I keep my lead } citrate (made up with ordinary distilled water, not specially } CO2 free) in a volumetric flask (50ml), which sits in the } same place month after month and is never moved. I don't use } the stain for 24 hours after it's prepared, but then just } carefully take off what's needed from close to the surface } using a glass pipette. I wipe the end of the pipette with a } tissue before dispensing the stain and then discard the first } drop. I put the drops onto Parafilm in a covered glass petri } dish. No need for NaOH pellets. Finally wash the grids for 5 } seconds in a gentle stream of water from a wash bottle. I } probably shouldn't admit this but I've had a bottle of stain } last over 2 years (the surface of the bottle becomes cloudy } with ppte) and still produce perfect results. } } Cheers, } } Diana
I use similar protocol, but I do use DI-} distilled-} boiled water and I do put NaOH pellets in the petry dish. Never tried to keep stain for 2 years, but for 6 month it works fine.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 371 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} ---------------------------------------------------------------------- } } ------ } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver On-Line Help
==============================Original Headers============================== 9, 23 -- From DusevichV-at-umkc.edu Wed May 3 09:18:33 2006 9, 23 -- Received: from kc-msxproto3.kc.umkc.edu (exchange.umkc.edu [134.193.44.10]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k43EIXjZ011623 9, 23 -- for {microscopy-at-msa.microscopy.com} ; Wed, 3 May 2006 09:18:33 -0500 9, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by kc-msxproto3.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Wed, 3 May 2006 09:18:33 -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: RE: [Microscopy] Re: TEM--Lead Citrate--HELP!! 9, 23 -- Date: Wed, 3 May 2006 09:18:32 -0500 9, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADC12-at-KC-MSX1.kc.umkc.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: [Microscopy] Re: TEM--Lead Citrate--HELP!! 9, 23 -- Thread-Index: AcZuUkF607FRWDc1TtCl4//Np/qEYQAaLHnw 9, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 9, 23 -- To: {microscopy-at-msa.microscopy.com} 9, 23 -- X-OriginalArrivalTime: 03 May 2006 14:18:33.0065 (UTC) FILETIME=[757CAD90:01C66EBC] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k43EIXjZ011623 ==============================End of - Headers==============================
I have been using "calcined lead citrate" (apparently a modification of Sato's lead citrate) for a couple of years, and it certainly is much more stable than traditional Reynold's lead citrate. Takamasa Hanaichi et al. (1986) A Stable Lead by Modification of Sato's Method. J. Electron Microsc., Vol. 35. No. 3. 304-306. --Jan Factor
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College, State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
mike.reedy-at-cellbio.duke.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 --| ---------------------------------------------------------------------------- --| --| Doesn't anybody else use or recommend Sato's lead stain as a more --| stable replacement for Reynolds Pb citrate? We've used it since the --| 1970s. --| 1968 Sato, T.: J. Electron Microsc. 17:158, 1968. --| 1968 Sato and others: Proc. XIth Int. Cong. on Electron Microscopy. --| Kyoto. 1986, pp. 2181-2182. [ --| --| -mike reedy- --| --| At 5:49 PM -0500 5/2/06, dcrippen-at-buckinstitute.org wrote: --| --|--| ---------------------------------------------------------------------------- --|--| The Microscopy ListServer -- CoSponsor: The Microscopy Society of America --|--| To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver --|--| On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html --|--| ---------------------------------------------------------------------------- --|--| --|--| Dear TEM users, --|--| --|--| This is a specimen prep question for everyone out there with EM --|--| expertise. We are having terrible success with Lead Citrate --|--| contrast staining. Principally, we suffer from precipitates showing --|--| up all over the specimen. On the advice of EM science technical --|--| support, we are double distilling our own water (they say Milli-Q is --|--| too pure and also is de-ionized which we don't want for EM). Then --|--| we make it CO2 free by autoclaving and capping directly upon removal --|--| --| --|from the autoclave. We do this the morning of reagent prep--so it --| --|--| doesn't sit in the bottle for longer than it takes to cool down --|--| before we begin making up the Reynolds. --|--| --|--| We make the Reynolds Lead citrate according to the protocol listed --|--| in Ch 5 of Bozzola and Russell's Electron Microscopy (2nd edition) --|--| (mix 1.33g lead nitrate, 1.76g sodium citrate, and 30ml CO2 free --|--| double distilled water...shake vigorously for a few miutes and then --|--| again 5-6 times over the next 30 minutes). Ensure solution is milky --|--| white and free of particles. Add 8.0 ml commercially prepared, --|--| titrated 1.0 N NaOH--from EM science--solution turns clear. Adjust --|--| pH strictly to 12.0+/-0.1 unit. Bring volume to 50ml with CO2-free --|--| double distilled water. Stopper tightly with rubber stopper and --|--| parafilm until use later that day. --|--| --|--| When we stain the grids with lead nitrate, we make sure to wash well --|--| before and after with CO2 free double distilled water in addition to --|--| surrounding the staining plate (Hiraoka kit) with NaOH pellets. In --|--| addition, we centrifuge the Reynolds at 5000xg for 8 minutes prior --|--| to use AND we 0.2um filter it into staining plate. --|--| --|--| ANY advice or thoughts are welcome...what are we doing wrong?? What --|--| can we change about this protocol to ensure ppt free staining? --|--| --|--| A thousand thanks in advance! --|--| --|--| Danielle Crippen --|--| Morphology and Imaging Core Manager --|--| Buck Institute for Age Research --|--| 8001 Redwood Blvd. --|--| Novato, CA 94945 --|--| 415-209-2046 --|--| dcrippen-at-buckinstitute.org --|--| --|--|
==============================Original Headers============================== 7, 16 -- From jfactor-at-ns.purchase.edu Wed May 3 17:27:11 2006 7, 16 -- Received: from zephyr.ns.purchase.edu (zephyr.ns.purchase.edu [199.79.168.193]) 7, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k43MRAVQ029989 7, 16 -- for {microscopy-at-microscopy.com} ; Wed, 3 May 2006 17:27:10 -0500 7, 16 -- Received: from [10.52.0.79] ([10.52.0.79]) 7, 16 -- by zephyr.ns.purchase.edu (AIX5.1/8.11.6p2/8.11.0) with ESMTP id k43MV1520666; 7, 16 -- Wed, 3 May 2006 18:31:01 -0400 7, 16 -- Message-ID: {44592E41.5000605-at-ns.purchase.edu} 7, 16 -- Date: Wed, 03 May 2006 18:27:13 -0400 7, 16 -- From: Jan Factor {jfactor-at-ns.purchase.edu} 7, 16 -- User-Agent: Thunderbird 1.5 (Windows/20051201) 7, 16 -- MIME-Version: 1.0 7, 16 -- To: microscopy-at-microscopy.com 7, 16 -- Subject: Re: Lead Citrate 7, 16 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 16 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I've had a couple of inquiries about calcined lead citrate, so I thought I'd send this to the list in case anyone else is interested. (To be fair, I learned about this method by perusing the EMS Catalog, which has the formulation.) I've pasted the relevant page from my in-house lab manual (below). To prepare the calcined the lead citrate, the unusual step, I simply went up to our chemistry program and asked them to fire up their high-temp oven (which is in a fume hood) for the day. Once you get a successful batch of calcined lead citrate, and I suggest making a good deal more than you need immediately, it can be stored as a powder in a vial for some time (perhaps indefinitely?). This way, you only have to bake it once, and you can make enough for multiple batches of lead citrate stain. I still use the usual precautions when handling lead stain, such as using NaOH pellets, and I spin down the stain in a table-top centrifuge before each use. Hope this is helpful. --Jan
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
Calcined lead citrate (Hanaichi et al., 1986)
Calcined lead citrate is a stable, non-precipitating replacement for Reynolds’ lead citrate, which is reportedly free from precipitates for over one year when kept at room temperature. Takamasa Hanaichi et al. (1986) A Stable Lead by Modification of Sato's Method. J. Electron Microsc., Vol. 35. No. 3. 304-306. Calcined lead citrate: Heat crystal lead citrate for several hours in a melting pot (200-300̊C) until the color changes to a light brownish yellow. This takes ~6.5 hrs at 250̊C. Note: Check the color periodically, as overheated lead citrate with a dark brownish or black color can't be used. The calcined lead citrate can be stored and used for repeated batches of stain.
The stock lead solution: 1. The following reagents are placed in a 50 ml volumetric flask and mixed well to produce a yellowish milky solution: CALCINED LEAD CITRATE........0.20 g Prepared from lead citrate Lead nitrate......................................0.15 g Lead acetate....................................0.15 g EMS #17600-25, 25g Sodium citrate.................................1.00 g Distilled water................................ 41.00 ml
2. Then, add: 1.0N NaOH.....................................9.0 ml Carbonate free, EMS #21170-01, 225ml Then 9.0 ml of 1N NaOH (use carbonate free solution) is added to the solution and mixed well until the solution becomes clear with a light yellowish color. The solution is then transferred to an amber glass with screw cap bottle for storage, and can be stored at room temperature or in the refrigerator (recommended) for over 1 year.
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==============================Original Headers============================== 12, 19 -- From jfactor-at-ns.purchase.edu Wed May 3 19:02:12 2006 12, 19 -- Received: from mta3.srv.hcvlny.cv.net (mta3.srv.hcvlny.cv.net [167.206.4.198]) 12, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4402Cd9008597 12, 19 -- for {microscopy-at-msa.microscopy.com} ; Wed, 3 May 2006 19:02:12 -0500 12, 19 -- Received: from [127.0.0.1] (ool-4357e602.dyn.optonline.net [67.87.230.2]) 12, 19 -- by mta3.srv.hcvlny.cv.net 12, 19 -- (Sun Java System Messaging Server 6.2-4.03 (built Sep 22 2005)) 12, 19 -- with ESMTP id {0IYP00FL8S34FY65-at-mta3.srv.hcvlny.cv.net} for 12, 19 -- microscopy-at-msa.microscopy.com; Wed, 03 May 2006 20:01:53 -0400 (EDT) 12, 19 -- Date: Wed, 03 May 2006 20:02:00 -0400 12, 19 -- From: Jan Factor {jfactor-at-ns.purchase.edu} 12, 19 -- Subject: Calcined lead citrate 12, 19 -- To: Microscopy Listserver {microscopy-at-msa.microscopy.com} 12, 19 -- Message-id: {44594478.8020707-at-ns.purchase.edu} 12, 19 -- MIME-version: 1.0 12, 19 -- Content-type: text/plain; charset=UTF-8; format=flowed 12, 19 -- User-Agent: Thunderbird 1.5.0.2 (Windows/20060308) 12, 19 -- Content-Transfer-Encoding: 8bit 12, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4402Cd9008597 ==============================End of - Headers==============================
I have a client who would like to do some confocal on GFP-transformed Arabidopsis (no problem) and then some TEM immunolabeling. What do I need to know about buying an anti-GFP antibody conjugated to gold? Any tricks, or is it straightforward? Does anyone have a favorite vendor? Any tips gratefully accepted!
Aloha, 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============================== 5, 19 -- From tina-at-pbrc.hawaii.edu Wed May 3 22:56:05 2006 5, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k443u4UL021598 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 22:56:05 -0500 5, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 5, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k443u0kp023231 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 17:56:00 -1000 (HST) 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k443txfE023228 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 17:55:59 -1000 (HST) 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 5, 19 -- Date: Wed, 3 May 2006 17:55:59 -1000 (HST) 5, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 5, 19 -- X-Sender: tina-at-halia 5, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 5, 19 -- Subject: Anti-GFP for TEM? 5, 19 -- Message-ID: {Pine.GSO.4.21.0605031752360.23000-100000-at-halia} 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
I am observing the internalization of a fluorescent component in the cells. I thought it would be a good idea to quench the extracellular fluorescence after several hours of incubation. This way I would see only the fluorescence coming from inside the cells. Would you know a substance which quenches Alexa488 dyes and which is not toxic to the cells (and do not enter the cells)? Stéphane-without-an-i
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==============================Original Headers============================== 3, 18 -- From nizets2-at-yahoo.com Thu May 4 05:44:45 2006 3, 18 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k44Aij0f004607 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 4 May 2006 05:44:45 -0500 3, 18 -- Received: (qmail 1686 invoked by uid 60001); 4 May 2006 10:44:45 -0000 3, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 3, 18 -- s=s1024; d=yahoo.com; 3, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 3, 18 -- b=nqm/XzURMI3axvbmJZsvwkIBQ/KWvp7vwWzV66J26ivZKNEbGL+t4EtML2piRt2x+fsuORpYb9WXbSAqyni8b1Ok2ZWbjYfcwVq3Fb/bXNgmLACLGSGYC7RTq4WHmfsNfE3pR0v7ZMhk/BbD3jqLwcQQOlilu+MaLkA0l22SBw8= ; 3, 18 -- Message-ID: {20060504104445.1684.qmail-at-web37413.mail.mud.yahoo.com} 3, 18 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Thu, 04 May 2006 03:44:45 PDT 3, 18 -- Date: Thu, 4 May 2006 03:44:45 -0700 (PDT) 3, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 3, 18 -- Subject: quenching extracellular fluorescence 3, 18 -- To: microscopy-at-microscopy.com 3, 18 -- MIME-Version: 1.0 3, 18 -- Content-Type: text/plain; charset=iso-8859-1 3, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I am not aware of anti-GFP gold conjugate (although it may be somewhere out there), but you can use the indirect method, i.e. primary antibody/secondary conjugate (protein A/gold or secondary antibody/gold). For the primary, we use Torrey Pines Biolabs purified rabbit Anti-GFP, that works well even after 2% formaldehyde/0.2% glutaraldehyde (that is what we use for Tokuyashu's technique).
Hope this helps,
Michal
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 would like to do some confocal on GFP-transformed } Arabidopsis (no problem) and then some TEM immunolabeling. What do I need } to know about buying an anti-GFP antibody conjugated to gold? Any tricks, } or is it straightforward? Does anyone have a favorite vendor? Any tips } gratefully accepted! } } Aloha, } 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============================== } 5, 19 -- From tina-at-pbrc.hawaii.edu Wed May 3 22:56:05 2006 } 5, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) } 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k443u4UL021598 } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 22:56:05 -0500 } 5, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) } 5, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k443u0kp023231 } 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 17:56:00 -1000 (HST) } 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k443txfE023228 } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 17:55:59 -1000 (HST) } 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs } 5, 19 -- Date: Wed, 3 May 2006 17:55:59 -1000 (HST) } 5, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } 5, 19 -- X-Sender: tina-at-halia } 5, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} } 5, 19 -- Subject: Anti-GFP for TEM? } 5, 19 -- Message-ID: {Pine.GSO.4.21.0605031752360.23000-100000-at-halia} } 5, 19 -- MIME-Version: 1.0 } 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII } ==============================End of - Headers============================== }
==============================Original Headers============================== 6, 20 -- From M_Jarnik-at-fccc.edu Thu May 4 07:20:26 2006 6, 20 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k44CKQZn015740 6, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 May 2006 07:20:26 -0500 6, 20 -- Received: from [131.249.8.162] (macloan2.fccc.edu [131.249.8.162]) 6, 20 -- by azah.fccc.edu (8.12.11/8.12.11) with ESMTP id k44CKPHb019420 6, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 May 2006 08:20:26 -0400 (EDT) 6, 20 -- Message-ID: {4459F2D9.5010705-at-fccc.edu} 6, 20 -- Date: Thu, 04 May 2006 08:26:01 -0400 6, 20 -- From: Michal Jarnik {M_Jarnik-at-fccc.edu} 6, 20 -- Reply-To: M_Jarnik-at-fccc.edu 6, 20 -- Organization: Fox Chase Cancer Center 6, 20 -- User-Agent: Thunderbird 1.5 (Macintosh/20051201) 6, 20 -- MIME-Version: 1.0 6, 20 -- To: Microscopy-at-MSA.Microscopy.Com 6, 20 -- Subject: Re: [Microscopy] Anti-GFP for TEM? 6, 20 -- References: {200605040356.k443usWD022646-at-ns.microscopy.com} 6, 20 -- In-Reply-To: {200605040356.k443usWD022646-at-ns.microscopy.com} 6, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Filtered] Re: [Microscopy] Anti-GFP for TEM?
Question: Hello Tina and Everyone:
We don't have a gold probe for this (yet), and can't comment on the best antibodies, but the question comes up occasionally, and we keep an eye out for references. We have reported on several that might be helpful in back issues of our newsletter:
http://www.nanoprobes.com/Newsletter_Archive.html
(1) Follet‚Gueye and co-workers have reported an improved fixation method for the immunogold localization of green fluorescent protein (GFP). The method is a simple one-step procedure which consists of fixing in the dark for 30 min, 60 min, or 120 min at 4C in a solution consisting of 1% glutaraldehyde and 1% osmium tetroxide in 0.1 M Na cacodylate buffer, pH 7.2. This mixture was prepared and used immediately. After washing in distilled water, the samples were gradually dehydrated in 10%, 20%, and 40% aqueous ethanol (10 min in each bath), then in 60% and 80% (20 min in each bath), and finally in anhydrous ethanol for 30 min, and embedded in LR White or Spurr resin. GFP-tagged Golgi glycosyltransferase was localized in transgenic BY-2 cells using rabbit anti-GFP primary and 10 nm gold anti-rabbit secondary antibodies; this method allowed improved structural preservation of the endomembrane system, and anti-GFP antibodies bound with high specificity, allowing the localization of GFP-tagged glycosyltransferases within individual Golgi cisternae.
Reference:
Follet-Gueye, M. L.; Pagny, S.; Faye, L.; Gomord, V., and Driouich, A.: An improved chemical fixation method suitable for immunogold localization of green fluorescent protein in the Golgi apparatus of tobacco Bright Yellow (BY-2) cells. J. Histochem. Cytochem., 51, 931-940 (2003).
Abstract (courtesy of the Journal of Histochemistry and Cytochemistry): http://www.jhc.org/cgi/content/abstract/51/7/931
Reprint (courtesy of the Journal of Histochemistry and Cytochemistry): http://www.jhc.org/cgi/reprint/51/7/931
(2) Luby-Phelps and co-workers describe a procedure for labeling green fluorescent protein (GFP)-expressing cells in 1-micron zebrafish sections for fluorescence and electron microscope observation using a 10 nm gold-labeled secondary antibody. GFP fluorescence survived fixation in 4% paraformaldehyde/0.1% glutaraldehyde and can be visualized directly by fluorescence microscopy. For electron microscopy, thin sections of silver‚gold color were collected on nickel grids and were incubated in anti-GFP antibody at a 1:25 dilution for 2 hr at room temperature, followed by 10-nm immunogold-conjugated goat anti-rabbit IgG for 1 hr. After contrasting with 3% aqueous uranyl acetate solution, the grids were dried and viewed with a Hitachi 600 transmission electron microscope operated at 75 kV.
Reference:
Luby‚Phelps, K.; Ning, G.; Fogerty, J., and Besharse, J. C.: Visualization of Identified GFP-expressing Cells by Light and Electron Microscopy. J. Histochem. Cytochem., 51, 271-274 (2003).
Abstract (courtesy of the Journal of Histochemistry and Cytochemistry): http://www.jhc.org/cgi/content/abstract/51/3/271
Reprint (courtesy of the Journal of Histochemistry and Cytochemistry): http://www.jhc.org/cgi/reprint/51/3/271
(3) Joanne Buchanan and co-workers investigated immuno-EM of GFP with NanogoldĆ secondary antibody conjugates, and described some results at Microscopy & Microanalysis 2002: use of microwave processing greatly shortened all the steps.
Reference:
Buchanan, J.; Micheva, K. D., and Smith, S. J.; Microwave Processing and Pre-embedding Nanogold Immunolabeling for Electron Microscopy. Microsc. Microanal., 8, (Suppl. 2: Proceedings); Lyman, C. E.; Albrecht, R. M.; Carter, C. B.; Dravid, V. P.; Herman, B., and Schatten, H. (Eds.); Cambridge University Press, New York, NY, 2002, 160.
Prior and co-workers prepared their own 5nm gold anti-GFP:
Prior, I. A.; Muncke, C.; Parton, R. G., and Hancock J. F.: Direct visualization of Ras proteins in spatially distinct cell surface microdomains. J. Cell. Biol., 160, 165-170 (2003).
Reprint (Journal of Cell Science): http://www.jcb.org/cgi/reprint/160/2/165
Hope some of this is helpful,
Rick Powell
******************************************************** Richard D. Powell rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated US Toll-free: (877) 447-6266 * Tel: (919) 845-6324 ********************************************************
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At 11:57 PM 5/3/2006, you wrote:
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Hi, All-
I have a client who would like to do some confocal on GFP-transformed Arabidopsis (no problem) and then some TEM immunolabeling. What do I need to know about buying an anti-GFP antibody conjugated to gold? Any tricks, or is it straightforward? Does anyone have a favorite vendor? Any tips gratefully accepted!
Aloha, 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* ****************************************************************************
Hi Tina, I have not had very good luck with localizing GFP for on TEM sectios. Hope you will share any good suggestions that you get
Greg
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 would like to do some confocal on GFP-transformed } Arabidopsis (no problem) and then some TEM immunolabeling. What do I need } to know about buying an anti-GFP antibody conjugated to gold? Any tricks, } or is it straightforward? Does anyone have a favorite vendor? Any tips } gratefully accepted! } } Aloha, } 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============================== } 5, 19 -- From tina-at-pbrc.hawaii.edu Wed May 3 22:56:05 2006 } 5, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) } 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k443u4UL021598 } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 22:56:05 -0500 } 5, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) } 5, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k443u0kp023231 } 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 17:56:00 -1000 (HST) } 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k443txfE023228 } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 May 2006 17:55:59 -1000 (HST) } 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs } 5, 19 -- Date: Wed, 3 May 2006 17:55:59 -1000 (HST) } 5, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } 5, 19 -- X-Sender: tina-at-halia } 5, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} } 5, 19 -- Subject: Anti-GFP for TEM? } 5, 19 -- Message-ID: {Pine.GSO.4.21.0605031752360.23000-100000-at-halia} } 5, 19 -- MIME-Version: 1.0 } 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII } ==============================End of - Headers============================== }
-- Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
-- Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 7, 24 -- From gwe-at-ufl.edu Thu May 4 10:17:48 2006 7, 24 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 7, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k44FHmNN006072 7, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 May 2006 10:17:48 -0500 7, 24 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 7, 24 -- (authenticated bits=0) 7, 24 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k44FHk3S1302702; 7, 24 -- Thu, 4 May 2006 11:17:46 -0400 7, 24 -- Message-ID: {445A1B19.3060508-at-ufl.edu} 7, 24 -- Date: Thu, 04 May 2006 11:17:45 -0400 7, 24 -- From: greg {gwe-at-ufl.edu} 7, 24 -- Reply-To: gwe-at-ufl.edu 7, 24 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 7, 24 -- X-Accept-Language: en-us, en 7, 24 -- MIME-Version: 1.0 7, 24 -- To: tina-at-pbrc.hawaii.edu, 7, 24 -- "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 7, 24 -- Subject: [Fwd: Re: [Microscopy] Anti-GFP for TEM?] 7, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 24 -- Content-Transfer-Encoding: 7bit 7, 24 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 7, 24 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 7, 24 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 7, 24 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Dear All, I am looking for a flange to mount a KEVEX EDS detector on my Philips 525 SEM (conical lens). Or for a drawing to help manufacturing the flange.
Hi Tina. We published a detailed protocol : Micheva, Buchanan, Hotz and Smith Nature Neuroscience 2003 Vol 6 #9 p925-32. I have had some good results with anti GFP antibodies from MBL and Roche for TEM. We used Fluro-Nanogold secondaries and silver enhanced and looked at hippocampal neurons in culture, Drosophila salivary glands and mouse brain -all pre-embedding labeling. I don't have any experience with aradadopsis. Good luck let me know if you need more info. JoAnn Buchanan
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
==============================Original Headers============================== 3, 18 -- From redhair-at-stanford.edu Thu May 4 13:09:06 2006 3, 18 -- Received: from smtp1.stanford.edu (smtp1.Stanford.EDU [171.67.22.28]) 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k44I964k029103 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 4 May 2006 13:09:06 -0500 3, 18 -- Received: from smtp1.stanford.edu (localhost.localdomain [127.0.0.1]) 3, 18 -- by localhost (Postfix) with SMTP id DDAF24BED1 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 4 May 2006 11:09:04 -0700 (PDT) 3, 18 -- Received: from Bucky.stanford.edu (B135-WinXP.Stanford.EDU [171.65.21.62]) 3, 18 -- by smtp1.stanford.edu (Postfix) with ESMTP id B47724C16D 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 4 May 2006 11:09:04 -0700 (PDT) 3, 18 -- Message-Id: {6.2.5.6.2.20060504110100.06cdd770-at-stanford.edu} 3, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 3, 18 -- Date: Thu, 04 May 2006 11:09:01 -0700 3, 18 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 3, 18 -- From: JoAnn Buchanan {redhair-at-stanford.edu} 3, 18 -- Subject: Anti-GFP for TEM 3, 18 -- Mime-Version: 1.0 3, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I am looking for a method to identify Type IIB skeletal muscle fibers at the electron microscopy level in human muscle biopsies. There are well established techniques for light microscopy, typically detecting ATPase, but I need good fixation and embedding for immunoelectron microscopy. A reasonable fixation compromise is 2% formaldehyde and 0.1% --0.2% glutaraldehyde with embedding in LR White/Gold or one of the Lowicryl resins. Any suggestions? Any experience with a particular antibody for human type IIB fibers?
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 4, 28 -- From Larry.Ackerman-at-ucsf.edu Thu May 4 14:35:17 2006 4, 28 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k44JZGEP007901 4, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 4 May 2006 14:35:16 -0500 4, 28 -- Received: from 64.54.128.152 by emfmcb02.ucsfmedicalcenter.org with 4, 28 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 4, 28 -- Thu, 04 May 2006 12:44:27 -0700 4, 28 -- X-Server-Uuid: 44B63953-633F-4500-B2DA-A3E478718104 4, 28 -- Received: from [128.218.123.88] ([128.218.123.88]) by 4, 28 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Thu, 4 May 4, 28 -- 2006 12:35:03 -0700 4, 28 -- Message-ID: {445A5767.7030601-at-ucsf.edu} 4, 28 -- Date: Thu, 04 May 2006 12:35:03 -0700 4, 28 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 4, 28 -- Reply-to: larry.ackerman-at-ucsf.edu 4, 28 -- Organization: UCSF, NeuroAnatomy 4, 28 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 4, 28 -- X-Accept-Language: en-us, en 4, 28 -- MIME-Version: 1.0 4, 28 -- To: Microscopy-at-microscopy.com 4, 28 -- Subject: Type IIB skeletal muscle 4, 28 -- X-OriginalArrivalTime: 04 May 2006 19:35:03.0740 (UTC) 4, 28 -- FILETIME=[D73977C0:01C66FB1] 4, 28 -- X-WSS-ID: 684486111G81226191-01-01 4, 28 -- Content-Type: text/plain; 4, 28 -- charset=iso-8859-1; 4, 28 -- format=flowed 4, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Email: bschneid-at-fhcrc.org Name: Bobbie S.
Organization: FHCRC
Title-Subject: [Filtered] Independent microscope service engineers for JEOL microsocpes
Question: Is there an independent person servicing JEOL microscopes who on the west coast,specifically Washington.
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 fsoheilian-at-ncifcrf.gov as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: fsoheilian-at-ncifcrf.gov Name: Ferri
Organization: NCI
Title-Subject: [Filtered] Breakage of section under the beam of TEM H7600 EM
Question: Dear All,
We're recently experiencing breakage of our sections (no matter what the thickness is) while imaging our samples under TEM H7600 Electron Microscope.
The section's thickness is usually 70-90 nm and spread nicely on a 200 mesh square Cu grid.
Our TEM H7600 is only two years old and works very well except for breaking the section. Even though we changed HV from 80 to 100 (Acc. Voltage Control, it didn't help and still broke the section.
We most likely experience this problem while imaging with the BottomMount of AMT542 camera, which used for high magnification samples.
Any comment on how to solve this problem or to control the beam from breaking the sections?
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Organization: Stockholm University, Stockholm, Sweden
Title-Subject: [Filtered] TEM EELS; any old/used spectrometer available?
Question: Hello everyone!
I have a question that I would like to ask and Gerald Kothleitner at Graz recommended this forum to me, i.e. this is my first posting here.
At Stockholm University, we are buying some new TEM and spectrometer equipment. In the purchase process, however, we have an extra microscope. My idea is, if possible, to buy an additional spectrometer, a used one, for example GIF or PEELS to use together with our extra microscope (our old JEOL 3010 LaB6 instrument) so that young scientists can get more access to the instruments as well. So my question is, if you have, or if you know someone who has an old GIF or PEELS or similar that is not or will not be needed anymore at your lab due to for example an upgrade, or if is not needed anymore for any reason. Then "I" would be very interested in somehow purchasing this old/used spectrometer system for an affordable price (because it is not part of our current budget). I am asking this question as a PhD student, but I will happily forward your proposals or give you the contact information to the people who are making the decisions at my university.
Dear Ferri, We have the same instrument and I have seen the problem you are experiencing. We have found it is usually a no more than a specimen clamping problem.You must make sure that the clamping mechanism is correctly located over the grid hole. We always use a pointed wooden spill to gently push it into place , you will hear a 'click' as it locates. It could be that the holder and clamping mechanism just need cleaning so that a good contact can be made.. This is not a unique problem to the H7600 in any TEM if the grid is not firmly clamped this will happen. Hope this solves it for you. Regards
Christine.
Christine Richardson Experimental Officer School of Biological and Biomedical Science Centre for Molecular Imaging University of Durham Science site South Rd Durham England DH1 3LE Tel: 0191 3341285\3341321 Fax:0191 3341201 E-mail: a.c.richardson-at-dur.ac.uk
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Thanks to all who responded with tips. We will try an indirect labeling process, and I will fix more lightly than usual. If we are dramatically successful, I'll post our procedure!
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 Fri May 5 13:54:00 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 k45IrxDX012977 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 5 May 2006 13:53:59 -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 k45IrtUn008540 6, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 5 May 2006 08:53:55 -1000 (HST) 6, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k45Irs2g008537 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 5 May 2006 08:53:54 -1000 (HST) 6, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 6, 19 -- Date: Fri, 5 May 2006 08:53:53 -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: Thanks- Anti-GFP for TEM 6, 19 -- Message-ID: {Pine.GSO.4.21.0605050850340.8459-100000-at-halia} 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Here is the May 2006 Microscopy Today table of contents. I will close the subscription list for this issue on Thursday May 11, 2006.
Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$50 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Ron Anderson, Editor =======================================================
Teaching Old Microscopes New Tricks Stephen W. Carmichael, Mayo Clinic
VisBio: a Flexible Open-Source Visualization Package for Multidimensional Image Data Curtis T. Rueden and Kevin W. Eliceiri, U. Wisconsin-Madison
BioImageXD – New Open Source Free Software for the Processing, Analysis and Visualization of Multidimensional Microscopic Images P. Kankaanpää1, K. Pahajoki1, V. Marjomäki2, J. Heino2, D. White2 1University Turku, 2University Jyväskylä, Finland
Novel Developments in High-Frequency Micro-Ultrasound Imaging Tom Little, VisualSonics Inc., Toronto, Ontario, Canada
Precise SEM Cross Section Polishing via Argon Beam Milling N. Erdman, R. Campbell, and S. Asahina,* JEOL USA Inc., Peabody, Massachusetts, *JEOL Ltd., Japan
Perfusion Fixation of Research Animals C. W. Scouten,* R. O’Connor,** & M. Cunningham** *MyNeuroLab.com, St Louis, MO, ** Harvard U., Belmont, MA
Three-Dimensional Crystallographic Analysis Beyond EBSD Mapping: The Next Dimension J.J.L. Mulders*, and A. Gholinia**, * FEI, Eindhoven, The Netherlands, ** HKL Technology, Hobro, Denmark
Improved Cutting of One-Micron Plastic Sections using Qwick Glass Knives J. T. McMahon and J. Alsouss, Cleveland Clinic Foundation, Ohio
The Scanning of Colour and B&W Film and Photographs for Image Processing, Analysis and Archiving - On a Tight Budget Keith J. Morris, The Institute of Ophthalmology, UCL, UK
A Comment on AFM vs. Replicas for High Resolution Imaging Don Chernoff, Advanced Surface Microscopy, Inc.
Embedding Cultured Cells Grown in Well Plates Leona Cohen-Gould, Cornell University, Ithaca, NY
Flies in a Box P. S. Connelly & L. Ruggiero,* *Oregon Health and Science U. Portland, OR
A Comment on using FLIM with FRET Karl Garsha, Roper Scientific, Tucson, AZ
Microscopy for Children Carolyn Schooley, MSA Project MICRO
New and Interesting at PITTCON & Industry News
NetNotes Topics
--LM - floaters
--SAMPLE PREPARATION - viral particles
--SAMPLE PREPARATION – cell culture preparation
--SAMPLE PREPARATION - propylene oxide
--SAMPLE PREPARATION - MgO preparation and Fe oxidation
--MICROTOMY – cleaning grids
--MICROTOMY - section thickness
--IMMUNOCYTOCHEMISTRY - nanogold in semithin sections
--EM - microscope cooling lines
--EM – operating voltage
--TEM - Replicas
--TEM - carbon post-coating
--EDS - Low Z peak pileup
Index of Advertisers
==============================Original Headers============================== 35, 18 -- From microscopytoday-at-tampabay.rr.com Fri May 5 14:51:40 2006 35, 18 -- Received: from ms-smtp-05.tampabay.rr.com (ms-smtp-05.tampabay.rr.com [65.32.5.135]) 35, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k45Jpej7023464 35, 18 -- for {Microscopy-at-Microscopy.Com} ; Fri, 5 May 2006 14:51:40 -0500 35, 18 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 35, 18 -- by ms-smtp-05.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k45JpZZh000413; 35, 18 -- Fri, 5 May 2006 15:51:38 -0400 (EDT) 35, 18 -- Message-ID: {445BACC4.1020406-at-tampabay.rr.com} 35, 18 -- Date: Fri, 05 May 2006 15:51:32 -0400 35, 18 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 35, 18 -- User-Agent: Thunderbird 1.5.0.2 (Windows/20060308) 35, 18 -- MIME-Version: 1.0 35, 18 -- To: Listserver {Microscopy-at-Microscopy.Com} , 35, 18 -- Confocal Listserver {confocal-at-listserv.buffalo.edu} 35, 18 -- Subject: Microscopy Today May 2006 Table of Contents 35, 18 -- Content-Type: text/plain; charset=windows-1252; format=flowed 35, 18 -- Content-Transfer-Encoding: 8bit 35, 18 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
I am new to the listserve and was hoping somebody could give me some advise and/or guidance. I am looking for the best way to embed, cut, and adhere to microscope slides wool/hair fibers. I have tried nitrocellulose/paraffin, Spurr's Resin, Epon, and LR white. I am having an issue with the fibers popping out of the resin during sectioning and/or during the sample processing. I have tried cutting the resin dry and wet - floating off into water. I have tried infiltrating with acetone/resin and ethanol/resin mixtures with the Spurr's resin, acetone/resin infiltration with the Epon and LR white. I seem to get the best cuts with LR white (5 microns) dry but then I have a problem adhering them to a slide for processing. To improve sample adhesion, I have tried albumin, gelatin, and neoprene. Of the three, neoprene seems to work the best. If anybody can offer advice for any or all of these issues, I would greatly appreciate it.
Thank you, Felicia Dixon
==============================Original Headers============================== 2, 16 -- From fmdixon-at-comcast.net Sat May 6 09:42:14 2006 2, 16 -- Received: from rwcrmhc13.comcast.net (rwcrmhc13.comcast.net [216.148.227.153]) 2, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k46EgEtL023443 2, 16 -- for {microscopy-at-microscopy.com} ; Sat, 6 May 2006 09:42:14 -0500 2, 16 -- Received: from rmailcenter06.comcast.net ([204.127.197.116]) 2, 16 -- by comcast.net (rwcrmhc13) with SMTP 2, 16 -- id {20060506144213m1300pbq3se} ; Sat, 6 May 2006 14:42:13 +0000 2, 16 -- Received: from [67.167.241.97] by rmailcenter06.comcast.net; 2, 16 -- Sat, 06 May 2006 14:42:13 +0000 2, 16 -- From: fmdixon-at-comcast.net 2, 16 -- To: microscopy-at-microscopy.com 2, 16 -- Subject: LM - hair sample prep 2, 16 -- Date: Sat, 06 May 2006 14:42:13 +0000 2, 16 -- Message-Id: {050620061442.29057.445CB5C500084791000071812200734830020198070B0300-at-comcast.net} 2, 16 -- X-Mailer: AT&T Message Center Version 1 (Apr 11 2006) 2, 16 -- X-Authenticated-Sender: Zm1kaXhvbkBjb21jYXN0Lm5ldA== ==============================End of - Headers==============================
If I recall my history of optics properly, people in the Middle Ages knew precisely what things looked like at the microscopical level but just couldn't draw accurate pictures of them. So they made the technological decision to develop the microscope so they could show each other what they already knew God had created. Really, it wasn't investigation, it was just confirmation of what they already knew was there. Thus, they were able to illustrate that ontogeny really does recapitulate philogeny. They "illustrated", not "investigated".
I hope we've settled this issue.
-Michael the OHR (Optics Historian of Record)
At 03:22 PM 04/20/06 -0500, you wrote:
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____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 11, 23 -- From cammer-at-aecom.yu.edu Mon May 8 10:23:27 2006 11, 23 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 11, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k48FNQJO025529 11, 23 -- for {microscopy-at-microscopy.com} ; Mon, 8 May 2006 10:23:27 -0500 11, 23 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 11, 23 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with SMTP id k48FNHvV019156 11, 23 -- for {microscopy-at-microscopy.com} ; Mon, 8 May 2006 11:23:24 -0400 11, 23 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 11, 23 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006050811232026666 11, 23 -- for {microscopy-at-microscopy.com} ; Mon, 08 May 2006 11:23:20 -0400 11, 23 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.30.137]) 11, 23 -- by post.aecom.yu.edu (Postfix) with ESMTP id F3B801A 11, 23 -- for {microscopy-at-microscopy.com} ; Mon, 8 May 2006 11:23:19 -0400 (EDT) 11, 23 -- Message-Id: {5.2.1.1.2.20060508111658.011768c8-at-mailserver.aecom.yu.edu} 11, 23 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 11, 23 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 11, 23 -- Date: Mon, 08 May 2006 11:23:24 -0400 11, 23 -- To: microscopy-at-microscopy.com 11, 23 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 11, 23 -- Subject: Re: Ethical question; investigation vs. illustration 11, 23 -- In-Reply-To: {200604202022.k3KKMNNG012581-at-ns.microscopy.com} 11, 23 -- Mime-Version: 1.0 11, 23 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
What!? There are several ideas in there that are hard for me to swallow. Of course, I know that my ability to swallow alone does not determine the truth of concept.
I am not so familiar with the early history of microscopy and the thinking surrounding it. (I would not think of challenging you for the position of OHR.) I doubt there was a decision made to develop microscopy to only verify what was suspected of being there. Their preconceptions might be considered hypotheses in need of verification, but we certainly get surprised enough about hypotheses on a macro scale. Microscopy would hardly be more of a sure thing.
I thought that microscopy would have developed more out of a desire to explore. I can guess what might be out there on a microscopic scale, but until I actually go and take a look, it is only conjecture - no matter how accurate the conjecture might be.
Maybe I am misunderstanding Mr. Cammer's point on Ontogeny recapitulating phylogeny. Are you saying that _those_ folks used microscopy to "prove" it? Perhaps they "proved" it to themselves. Or are you saying you still accept the idea yourself? If so, my understanding is that the concept has been discredited for some time. I am not a great fan of Stephen Gould but I found the following comment in an Amazon review of his book, _Ontogeny and Phylogeny_.
"Ontogeny recapitulates phylogeny" was Haeckel's answer--the wrong one--to the most vexing question of nineteenth-century biology: what is the relationship between individual development (ontogeny) and the evolution of species and lineages (phylogeny)? In this, the first major book on the subject in fifty years, Stephen Gould documents the history of the idea of recapitulation from its first appearance among the pre-Socratics to its fall in the early twentieth century. -Ernst Mayr.
FWIW, I consider microscopy an important tool for investigation. However, it must be used carefully as many of our observations are quite few. We need to pay attention to the statistics if we are going to generalize.
Warren Straszheim Iowa State University
-----Original Message----- X-from: cammer-at-aecom.yu.edu [mailto:cammer-at-aecom.yu.edu] Sent: Monday, May 08, 2006 10:26 AM To: wesaia-at-iastate.edu
Microscopists, Please please forgive me for leaving out the word "only" in my original post. I meant to write ..."it is worth remembering that microscopy can be used for demonstration not ONLY investigation.
I was simply trying to point out that it is reasonable to demand one kind of thing when we are investigating but also have room for another purpose, namely the demonstration, for which demands are different. In no way shape or form was I meaning to suggest that microscopy cannot be used to investigate. Indeed, that suggestion is ludicrous.
I did send an apology after my post and it seems once is not enough. So once more, I am sorry to have wasted time and bandwith with my failure to proofread.
As ever, Tobias
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I agree that EM or LM can and are used for investigation. They are also used for discovery. In the context of IC reverse engineering and technology evaluation, discovery and investigation are biggies. For failure analysis, I suspect that investigation fits the bill better than does discovery--unless investigation leads to the discovery of the failure mechanism.
IMO, I suspect that the early scientists and researchers had a clue that microbes existed but did not know what they looked like. Along comes the microscope. Now they could get a good idea of what they looked like and confirm that they existed. Is this investigation, discovery or both?
gary g.
At 10:23 AM 5/8/2006, you wrote:
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==============================Original Headers============================== 11, 21 -- From gary-at-gaugler.com Mon May 8 13:01:33 2006 11, 21 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k48I1Xf1025048 11, 21 -- for {microscopy-at-microscopy.com} ; Mon, 8 May 2006 13:01:33 -0500 11, 21 -- Received: (qmail 29671 invoked from network); 8 May 2006 11:01:32 -0700 11, 21 -- Received: by simscan 1.1.0 ppid: 29668, pid: 29669, t: 0.2843s 11, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 21 -- by qsmtp2 with SMTP; 8 May 2006 11:01:32 -0700 11, 21 -- Message-Id: {7.0.1.0.2.20060508105621.024f5678-at-gaugler.com} 11, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 21 -- Date: Mon, 08 May 2006 11:01:34 -0700 11, 21 -- To: baskin-at-bio.umass.edu 11, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 21 -- Subject: Re: [Microscopy] Re: Ethical question; investigation vs. 11, 21 -- illustration 11, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 21 -- In-Reply-To: {200605081723.k48HND4g017893-at-ns.microscopy.com} 11, 21 -- References: {200605081723.k48HND4g017893-at-ns.microscopy.com} 11, 21 -- Mime-Version: 1.0 11, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
You're too clever by half. I assume that "philogeny" was not a misspelling.
Now, where did I hear that ontology recapitulates phylogeny?
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 } ---------------------------------------------------------------------- } ------ } } Microscopy can be used for investigation? Really? } } If I recall my history of optics properly, people in the Middle Ages } knew precisely what things looked like at the microscopical level but } just couldn't draw accurate pictures of them. So they made the } technological decision to develop the microscope so they could show } each other what they already knew God had created. Really, it wasn't } investigation, it was just confirmation of what they already knew was } there. Thus, they were able to illustrate that ontogeny really does } recapitulate philogeny. They "illustrated", not "investigated". } } I hope we've settled this issue. } } -Michael the OHR (Optics Historian of Record) } } } At 03:22 PM 04/20/06 -0500, you wrote: } } } } } --------------------------------------------------------------------- } } ------- The Microscopy ListServer -- CoSponsor: The Microscopy } } Society of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver On-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html
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============================== 7, 27 -- From jbs-at-temple.edu Mon May 8 13:04:33 2006 7, 27 -- Received: from po-smtp3.temple.edu (po-smtp3.temple.edu [155.247.166.231]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k48I4XhC028812 7, 27 -- for {microscopy-at-microscopy.com} ; Mon, 8 May 2006 13:04:33 -0500 7, 27 -- Received: from [155.247.98.40] (jbs.bio.temple.edu [155.247.98.40]) 7, 27 -- by po-smtp3.temple.edu (MOS 3.7.5-GA) 7, 27 -- with ESMTP id CPU52947 (AUTH jbs); 7, 27 -- Mon, 8 May 2006 14:04:08 -0400 (EDT) 7, 27 -- From: "Joel Sheffield" {jbs-at-temple.edu} 7, 27 -- To: cammer-at-aecom.yu.edu, cammer-at-aecom.yu.edu, microscopy-at-microscopy.com 7, 27 -- Date: Mon, 08 May 2006 14:04:41 -0400 7, 27 -- MIME-Version: 1.0 7, 27 -- Subject: Re: [Microscopy] Re: Ethical question; investigation vs. illustration 7, 27 -- Reply-to: jbs-at-temple.edu 7, 27 -- Message-ID: {445F4FF9.1379.3AFE2A1D-at-jbs.temple.edu} 7, 27 -- Priority: normal 7, 27 -- In-reply-to: {200605081524.k48FOBBI025741-at-ns.microscopy.com} 7, 27 -- X-mailer: Pegasus Mail for Windows (4.30 public beta 1) 7, 27 -- Content-type: text/plain; charset=US-ASCII 7, 27 -- Content-transfer-encoding: 7BIT 7, 27 -- Content-description: Mail message body 7, 27 -- X-Junkmail-Status: score=10/50, host=po-smtp3.temple.edu 7, 27 -- X-Junkmail-SD-Raw: score=unknown, 7, 27 -- refid=str=0001.0A090209.445F8635.0065,ss=1,fgs=0, 7, 27 -- ip=155.247.98.40, 7, 27 -- so=2006-03-30 10:46:40, 7, 27 -- dmn=5.1.5/2006-04-27 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both dotys-at-hss.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dotys-at-hss.edu Name: Steve Doty
Organization: Hospital for Special Surgery
Title-Subject: [Filtered] Billing for core services.
Question: As director of a microscopy core, I am also responsible for sending bills/invoices for services provided by core. We have external as well as internal users. We provide TEM, SEM, LM, morphometric analysis, paraffin embedding and methacrylate embedding and sectioning, immuno and enzyme histochemistry. And we have a mix of clinical and basic research to accomodate. My problem is trying to keep the user and techniques together for billing purposes since any particular technique may take several days and different technician's support. Does anyone have a "system" that will help me keep track of everything that is happening around me! Many thanks, Steve Doty, PhD.
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 crnl_srbu-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Organization: Natl.Inst.for Materials Physics, Bucharest, Romania
Title-Subject: [Filtered] Zr70Ni10Pd20 metalic glasses - how can they be acid dissolved
Question: Dear colleagues,
Is there anybody who could give me a hint concerning the acid or acid mixture that would be effective in surface etching a piece of ternary metallic glass having the composition Zr70Ni10Pd20 ? I need to reveal the dimensions of grains which are most probably formed in the material after a short aannealing during which an icosahedral quasicrystalline phase was formed (it was revealed by X-ray diffraction). Any suggestion will be very welcome.
Thank you in advance.
Dr. Corneliu Sarbu National Institute for Materials Physics Magurele-Bucharest Romania
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 dainis-at-red5wood.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Although we are an academic lab in materials, our problems with billing are very similar. The data, of course, has to feed into the corporate financial management system (SAP, in our case).
After a long period of wondering what to do, and a false start with an almost institute-wide system that promised to be totally comprehensive but was in fact far too ambitious for our resources, we are currently developing an in-house system which will enable us to track usage by instrument, user, date, project, etc. etc., as well, of course, as providing the data for our billing.
It has a MySQL heart, with Visual Basic and Filemaker Pro front-ends (for historical reasons). The data collection part of the system is up and running, and in use for most of our systems. All the instruments should be using it by the end of next month. Then we have to develop the interface to the billing system, and the analysis tools (currently we extract the data from the database by manual SQL calls and generate a text file).
Tony
At 08:40 PM 5/8/2006, you wrote: Email: dotys-at-hss.edu Name: Steve Doty
Organization: Hospital for Special Surgery
Title-Subject: [Filtered] Billing for core services.
Question: As director of a microscopy core, I am also responsible for sending bills/invoices for services provided by core. We have external as well as internal users. We provide TEM, SEM, LM, morphometric analysis, paraffin embedding and methacrylate embedding and sectioning, immuno and enzyme histochemistry. And we have a mix of clinical and basic research to accomodate. My problem is trying to keep the user and techniques together for billing purposes since any particular technique may take several days and different technician's support. Does anyone have a "system" that will help me keep track of everything that is happening around me! Many thanks, Steve Doty, PhD.
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Good day, Corneliu, Considering Ti as chemically similar to Zr, I found some etchant solutions for Ti alloys in Metals Handbook from ASM. That these might work for Zr alloys is supported by comments in Cotton and Wilkinson, Adv. Inorg. Chem. Anyway, here are a couple that ASM suggests as general purpose etchants:
10 ml HF, 5 ml HNO3, 85ml water 1-3 ml HF, 2-6 ml HNO3, water to 1000 ml
You can also try contacting ATI Wah Chang in Oregon, USA. They've been producing Zr alloys for decades and have plenty of expertise in that area. phone: 1-541-926-4211 www.wahchang.com
Good luck, and BE CAREFUL if you use HF.
Rob Bowen
-- Robert C. Bowen Research Scientist Caddock Electronics, Inc rob.bowen-at-caddock.com http://www.caddock.com
} From: {crnl_srbu-at-yahoo.com} } Reply-To: {crnl_srbu-at-yahoo.com} } Date: Mon, 8 May 2006 19:37:44 -0500 } To: {rob.bowen-at-caddock.com} } Subject: [Microscopy] viaWWW: Zr70Ni10Pd20 metalic glasses - how can they be } acid } } } } } ---------------------------------------------------------------------------- } 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 crnl_srbu-at-yahoo.com as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: crnl_srbu-at-yahoo.com } Name: Corneliu Sarbu } } Organization: Natl.Inst.for Materials Physics, Bucharest, Romania } } Title-Subject: [Filtered] Zr70Ni10Pd20 metalic glasses - how can they be acid } dissolved } } Question: Dear colleagues, } } Is there anybody who could give me a hint concerning the acid or acid mixture } that would be effective in surface etching a piece of ternary metallic glass } having the composition Zr70Ni10Pd20 ? I need to reveal the dimensions of } grains which are most probably formed in the material after a short aannealing } during which an icosahedral quasicrystalline phase was formed (it was revealed } by X-ray diffraction). Any suggestion will be very welcome. } } Thank you in advance. } } Dr. Corneliu Sarbu } National Institute for Materials Physics } Magurele-Bucharest } Romania } } e-mail: crnl_srbu-at-yahoo.com } } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 11, 13 -- From zaluzec-at-microscopy.com Mon May 8 19:32:38 2006 } 11, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 11, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k490WbBA031859 } 11, 13 -- for {microscopy-at-microscopy.com} ; Mon, 8 May 2006 19:32:38 -0500 } 11, 13 -- Mime-Version: 1.0 } 11, 13 -- X-Sender: (Unverified) } 11, 13 -- Message-Id: {p06110403c0859396e5cc-at-[206.69.208.22]} } 11, 13 -- Date: Mon, 8 May 2006 19:32:36 -0500 } 11, 13 -- To: microscopy-at-microscopy.com } 11, 13 -- From: crnl_srbu-at-yahoo.com (by way of MicroscopyListserver) } 11, 13 -- Subject: viaWWW: Zr70Ni10Pd20 metalic glasses - how can they be acid } 11, 13 -- dissolved } 11, 13 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
==============================Original Headers============================== 11, 20 -- From Rob.Bowen-at-caddock.com Tue May 9 10:39:16 2006 11, 20 -- Received: from msg.caddock.com (69-29-3-221.stat.centurytel.net [69.29.3.221]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k49FdFhP022433 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 9 May 2006 10:39:16 -0500 11, 20 -- Received: from [10.1.2.107] ([10.1.2.107]) 11, 20 -- by msg.caddock.com (Merak 8.3.8) with ESMTP id NUW63907; 11, 20 -- Tue, 9 May 2006 08:39:07 -0700 11, 20 -- User-Agent: Microsoft-Entourage/11.0.0.040405 11, 20 -- Date: Tue, 09 May 2006 08:32:54 -0700 11, 20 -- Subject: Re: [Microscopy] viaWWW: Zr70Ni10Pd20 metalic glasses - how can 11, 20 -- they be acid 11, 20 -- From: Rob Bowen {Rob.Bowen-at-caddock.com} 11, 20 -- To: {crnl_srbu-at-yahoo.com} 11, 20 -- CC: {microscopy-at-microscopy.com} 11, 20 -- Message-ID: {C0860436.2FF3%Rob.Bowen-at-caddock.com} 11, 20 -- In-Reply-To: {200605090037.k490biw6016005-at-ns.microscopy.com} 11, 20 -- Mime-version: 1.0 11, 20 -- Content-type: text/plain; 11, 20 -- charset="US-ASCII" 11, 20 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
Hi, Might any of you know if polyurethane (Festo blue PU) tubing is compatible with P-10 gas. I currently have blue PU 3/16" tubing plumbed from the P-10 tank regulator to the first gfpc WDS on my uProbe. Is anyone 100% sure that this type of tubing should not be used to delivered P-10 gas to gas floow proportional counter detectors?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ******************************************************************** --
==============================Original Headers============================== 4, 12 -- From mmcheath-at-mailbox.syr.edu Tue May 9 16:45:07 2006 4, 12 -- Received: from mailer.syr.edu (mailer.syr.edu [128.230.18.29]) 4, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k49Lj7e5001406 4, 12 -- for {microscopy-at-ns.microscopy.com} ; Tue, 9 May 2006 16:45:07 -0500 4, 12 -- Received: from [128.230.24.90] (www.geochemistry.syr.edu) by mailer.syr.edu (LSMTP for Windows NT v1.1b) with SMTP id {0.1534C967-at-mailer.syr.edu} ; Tue, 9 May 2006 16:25:54 -0400 4, 12 -- Mime-Version: 1.0 4, 12 -- Message-Id: {a06230902c086aa40f639-at-[128.230.24.90]} 4, 12 -- Date: Tue, 9 May 2006 16:25:52 -0400 4, 12 -- To: microscopy-at-ns.microscopy.com 4, 12 -- From: Michael Cheatham {mmcheath-at-mailbox.syr.edu} 4, 12 -- Subject: [Microscopy] EPMA: P-10 gas and polyurethane tubing 4, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
CSIRO Plant Industry Black Mountain, Canberra Australia
Reference: 2006/411
We require an experienced microscopist with demonstrated knowledge of plant structure and developmental biology to assist in the Microscopy Centre in the Division of Plant Industry. Ideally, the appointee would have skills in preparation of plant material for light and electron microscopy, especially in cryo-scanning electron microscopy and x-ray microanalysis. The Microscopy Centre currently has light, fluorescence and confocal microscopes, a cryo-SEM with EDX, image analysis software and other ancilliary equipment.
The successful applicant will assist the Manager in training other staff in use of the instruments and in microscopy techniques, and in carrying out microscopy work for specific research projects. He/she will have at least a Bachelor's degree or equivalent training in plant structure and functional plant anatomy and experience in working in a research laboratory. Additional skills required are the ability to collaborate effectively with scientists and members of a research team and strong communication and computer skills.
This position is indefinite after a probationary period, salary $44K - $57K p.a plus Superannuation . To obtain selection documentation or details on how to apply visit http://recruitment.csiro.au/asp/job_details.asp?RefNo=2006%2F411. For further information contact Dr Rosemary White - rosemary.white-at-csiro.au . Responses to the selection criteria accompanied by a CV, must be received by close of business 4 June 2006.
Unfortunately, the powers-that-be have decided it's open to Australian residents only, largely so they don't have to fly people in for interviews. However, if you're interested, or know anyone who might be, contact me anyway.
cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au CSIRO Plant Industry ph. 02-6246 5475 GPO Box 1600 mob. 0402 835 973 Canberra, ACT 2601 fax. 02-6246 5334 Australia
==============================Original Headers============================== 6, 21 -- From Rosemary.White-at-csiro.au Tue May 9 22:16:44 2006 6, 21 -- Received: from act-MTAout4.csiro.au (act-MTAout4.csiro.au [150.229.7.41]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4A3Gfjj025723 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 9 May 2006 22:16:43 -0500 6, 21 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=bC6HYyfTUYh9CaPTvDUc3AjFXzW/zu8QZjKb+0k9l/CW8i5tvKSN+QHvOhErmGLrka3ACr9POWuOgf3jKrLizs3RXMNq5812A6CeLM2dzlUjk76BvvPASlMoyhkE+SAC; 6, 21 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 6, 21 -- by act-MTAout4.csiro.au with ESMTP; 10 May 2006 13:16:39 +1000 6, 21 -- X-IronPort-AV: i="4.05,107,1146405600"; 6, 21 -- d="scan'208"; a="93509110:sNHT29987620" 6, 21 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 6, 21 -- Wed, 10 May 2006 13:16:38 +1000 6, 21 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 21 -- Date: Wed, 10 May 2006 13:18:17 +1000 6, 21 -- Subject: position available 6, 21 -- From: Rosemary White {Rosemary.White-at-csiro.au} 6, 21 -- To: {Microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C0879899.16529%Rosemary.White-at-csiro.au} 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 10 May 2006 03:16:38.0982 (UTC) FILETIME=[26F0C660:01C673E0] ==============================End of - Headers==============================
We are considering buying EDX system integrated with EBSD for elemental and "structural" analysis of inorganic materials in SEM. I hope therefore that vendors of such equipment will contact me off the list. However, I would be grateful also for any comments from listers on the possibilities of EBSD as the method of structure identification of individual nanocrystals in composite materials.
Thank you,
Leszek
Leszek Kepinski Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland e-mail: L.Kepinski-at-int.pan.wroc.pl
==============================Original Headers============================== 13, 33 -- From L.Kepinski-at-int.pan.wroc.pl Wed May 10 05:31:44 2006 13, 33 -- Received: from mserv2.int.pan.wroc.pl (mserv2.int.pan.wroc.pl [156.17.85.6]) 13, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4AAVh7k009241 13, 33 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 05:31:43 -0500 13, 33 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 13, 33 -- by mserv2.int.pan.wroc.pl (INTiBS (1.0)) with ESMTP id 7143C134196 13, 33 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 12:43:45 +0200 (CEST) 13, 33 -- Received: from mserv2.int.pan.wroc.pl ([127.0.0.1]) 13, 33 -- by localhost (mserv2.int.pan.wroc.pl [127.0.0.1]) (amavisd-new, port 10025) 13, 33 -- with LMTP id 06540-02-6 for {Microscopy-at-microscopy.com} ; 13, 33 -- Wed, 10 May 2006 12:43:43 +0200 (CEST) 13, 33 -- X-Virus-Scanner: This message was checked by NOD32 Antivirus system 13, 33 -- NOD32 for Linux Mail Server. 13, 33 -- For more information on NOD32 Antivirus System, 13, 33 -- please, visit our website: http://www.nod32.com/. 13, 33 -- Received: from b8p020 (sqd.int.pan.wroc.pl [156.17.85.20]) 13, 33 -- by mserv2.int.pan.wroc.pl (INTiBS (1.0)) with SMTP id 2ED7B134276 13, 33 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 12:36:15 +0200 (CEST) 13, 33 -- Message-ID: {006101c6741b$d020fe40$7f01a8c0-at-b8p020} 13, 33 -- From: =?iso-8859-2?B?TC4gS+pwafFza2k=?= {L.Kepinski-at-int.pan.wroc.pl} 13, 33 -- To: {Microscopy-at-microscopy.com} 13, 33 -- Subject: EDS_EBSD system 13, 33 -- Date: Wed, 10 May 2006 12:23:43 +0200 13, 33 -- MIME-Version: 1.0 13, 33 -- Content-Type: text/plain; 13, 33 -- charset="iso-8859-2" 13, 33 -- Content-Transfer-Encoding: 7bit 13, 33 -- X-Priority: 3 13, 33 -- X-MSMail-Priority: Normal 13, 33 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 13, 33 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 13, 33 -- X-Scan-Module: SMTP[2006.04.28 (2006.01.25)] 13, 33 -- X-Virus-Scanned: by amavisd-new at int.pan.wroc.pl ==============================End of - Headers==============================
Hello, I'm using an older Pelco CPD 2 critical point drier. Anyone use this? It has these gnurled steel knobs for opening/closing the valves. Controlling the valves kills your fingers since the rough metal is hard on your skin. Anyone modified it with plastic knobs or rubber covers or something to make it less of a literal pain to do a critical point drying run?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Wed May 10 18:04:34 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 k4AN4YCx001108 2, 24 -- for {microscopy-at-microscopy.com} ; Wed, 10 May 2006 18:04:34 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 26521C1E40 2, 24 -- for {microscopy-at-microscopy.com} ; Wed, 10 May 2006 16:04:34 -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 02522-01 for {microscopy-at-microscopy.com} ; 2, 24 -- Wed, 10 May 2006 16:04:23 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id F2661C1E4F; Wed, 10 May 2006 16:04:22 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id EB150C1E43 2, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 16:04:22 -0700 (PDT) 2, 24 -- Date: Wed, 10 May 2006 16:04:22 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: Microscopy-at-microscopy.com 2, 24 -- Subject: cpd pains 2, 24 -- Message-ID: {Pine.SOC.4.64.0605101602510.1811-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==============================
When we had one of those we used pliers to turn the knobs.
gvrdolja-at-nature.berkeley.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello, } I'm using an older Pelco CPD 2 critical point drier. Anyone use this? It } has these gnurled steel knobs for opening/closing the valves. Controlling } the valves kills your fingers since the rough metal is hard on your skin. } Anyone modified it with plastic knobs or rubber covers or something to } make it less of a literal pain to do a critical point drying run? } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } 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 Wed May 10 18:04:34 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 k4AN4YCx001108 } 2, 24 -- for {microscopy-at-microscopy.com} ; Wed, 10 May 2006 18:04:34 -0500 } 2, 24 -- Received: from localhost (localhost [127.0.0.1]) } 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 26521C1E40 } 2, 24 -- for {microscopy-at-microscopy.com} ; Wed, 10 May 2006 16:04:34 -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 02522-01 for {microscopy-at-microscopy.com} ; } 2, 24 -- Wed, 10 May 2006 16:04:23 -0700 (PDT) } 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) } 2, 24 -- id F2661C1E4F; Wed, 10 May 2006 16:04:22 -0700 (PDT) } 2, 24 -- Received: from localhost (localhost [127.0.0.1]) } 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id EB150C1E43 } 2, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 16:04:22 -0700 (PDT) } 2, 24 -- Date: Wed, 10 May 2006 16:04:22 -0700 (PDT) } 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} } 2, 24 -- To: Microscopy-at-microscopy.com } 2, 24 -- Subject: cpd pains } 2, 24 -- Message-ID: {Pine.SOC.4.64.0605101602510.1811-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============================== }
-- Greg Erdos 5410 SE 185th Ave. Micanopy, FL 32667
==============================Original Headers============================== 3, 24 -- From gwe-at-ufl.edu Wed May 10 20:59:01 2006 3, 24 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4B1x0jY013536 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 10 May 2006 20:59:00 -0500 3, 24 -- Received: from [10.228.8.36] (ssrb-vpn2-8-36.vpn.ufl.edu [10.228.8.36]) 3, 24 -- (authenticated bits=0) 3, 24 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k4B1wvei508066; 3, 24 -- Wed, 10 May 2006 21:58:58 -0400 3, 24 -- Message-ID: {44629A6A.7050902-at-ufl.edu} 3, 24 -- Date: Wed, 10 May 2006 21:59:06 -0400 3, 24 -- From: greg erdos {gwe-at-ufl.edu} 3, 24 -- Reply-To: gwe-at-ufl.edu 3, 24 -- User-Agent: Thunderbird 1.5.0.2 (Windows/20060308) 3, 24 -- MIME-Version: 1.0 3, 24 -- To: gvrdolja-at-nature.berkeley.edu, microscopy-at-microscopy.com 3, 24 -- Subject: Re: [Microscopy] cpd pains 3, 24 -- References: {200605102306.k4AN68TT002400-at-ns.microscopy.com} 3, 24 -- In-Reply-To: {200605102306.k4AN68TT002400-at-ns.microscopy.com} 3, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 24 -- Content-Transfer-Encoding: 7bit 3, 24 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 24 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 24 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 24 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Sending to list--direct msgs bounce. If you have a better address, please advise and I will send any other material off-list.
-------
What SEM do you plan on using? What type of gun does it use?
What feature sizes are you interested in resolving? Ta and Cu seed layers are not possible to resolve even at .01u step size. I think that this is because no discernable grains have formed as yet. The electrodep Cu for damascene interconnects works well.
I've done single crystal Sapphire, Silicon and micro crystal Si. What would your nano crystals be made of? If the material is non-conductive, this may pose a charging issue. Since EBSD penetrates only about 50nm, this does not leave much depth for coating. The preferred coating is C. However, I rarely coat any insulating material even at 20KV.
There are basically two EBSD choices. TSL/EDAX or HKL/PGT. TSL and EDAX have been integrated for much longer than HKL and PGT (I think HKL connected just this year). TSL's camera/phosphor is round while HKL's camera end is square. This allows the HKL camera screen to get closer to the specimen. But I think that the TSL software complement is much better. AFAIK, only TSL has drift correction during data collection. At high mag and small step size, this is critical. However TSL's drift correction is not always repeatable and its own set of problems. But at least it is there and does usually work.
The SEM is going to be at issue too since higher frames per second need more probe current. However, this is at the expense of probe diameter and lattice resolution.
I'm using EDAX/TSL Pegasus with EDAX Genesis EDS and like it. The SEM is a Zeiss Supra 55VP, which has its own set of issues.
gary g.
At 03:34 AM 5/10/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 15, 19 -- From gary-at-gaugler.com Thu May 11 11:17:49 2006 15, 19 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 15, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4BGHmMP004357 15, 19 -- for {microscopy-at-microscopy.com} ; Thu, 11 May 2006 11:17:48 -0500 15, 19 -- Received: (qmail 26316 invoked from network); 11 May 2006 09:17:46 -0700 15, 19 -- Received: by simscan 1.1.0 ppid: 26311, pid: 26312, t: 0.1909s 15, 19 -- scanners: regex: 1.1.0 attach: 1.1.0 15, 19 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 15, 19 -- by qsmtp3 with SMTP; 11 May 2006 09:17:46 -0700 15, 19 -- Message-Id: {7.0.1.0.2.20060511090631.02476f70-at-gaugler.com} 15, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 19 -- Date: Thu, 11 May 2006 09:17:46 -0700 15, 19 -- To: MSA listserver {microscopy-at-microscopy.com} 15, 19 -- From: Gary Gaugler {gary-at-gaugler.com} 15, 19 -- Subject: Re: [Microscopy] EDS_EBSD system 15, 19 -- In-Reply-To: {200605101034.k4AAYo4n012418-at-ns.microscopy.com} 15, 19 -- References: {200605101034.k4AAYo4n012418-at-ns.microscopy.com} 15, 19 -- Mime-Version: 1.0 15, 19 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Hi Gordon - we have that same unit and have well developed calluses! Two simple tricks to keep your skin more or less intact: - I never adjust the coarse vent valve. Permanently leave it halfway open and control venting with the needle valve. - When going through the fill-drain cycles - open the fill valve one-half turn or so and leave it there until you're finished. Then you just open the drain valve to let fluid out, and close it to let fluid in. This cuts the number of knob-turning operations down by half. On our unit, the fill valve is the hardest to operate so this trick is very helpful.
I'd avoid pliers unless you absolutely can't get the knobs to turn. Pliers can strip the knurled texture and just make things harder in the future. And too much force can damage the valves. I do have one user that uses vise-grip pliers on the fill valve, but she is under stern warnings to pad the knob with cloth and not use force to close the valve.
Rick
gvrdolja-at-nature.berkeley.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello, } I'm using an older Pelco CPD 2 critical point drier. Anyone use this? It } has these gnurled steel knobs for opening/closing the valves. Controlling } the valves kills your fingers since the rough metal is hard on your skin. } Anyone modified it with plastic knobs or rubber covers or something to } make it less of a literal pain to do a critical point drying run? } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } 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 } }
-- {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Richard C. Hugo, Ph.D Geomicrobiology and Electron Microscopy Laboratory Portland State University Ph# 503-725-3356 FAX 503-725-3025 {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
==============================Original Headers============================== 6, 23 -- From hugo-at-pdx.edu Thu May 11 11:40:54 2006 6, 23 -- Received: from fafnir.oit.pdx.edu (fafnir.oit.pdx.edu [131.252.120.58]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4BGerPO014461 6, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 11 May 2006 11:40:53 -0500 6, 23 -- Received: from [131.252.193.7] (host-193-7.dhcp.pdx.edu [131.252.193.7]) 6, 23 -- (authenticated bits=0) 6, 23 -- by fafnir.oit.pdx.edu (8.13.3+/8.13.1) with ESMTP id k4BGeo6r025730 6, 23 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 6, 23 -- Thu, 11 May 2006 09:40:51 -0700 6, 23 -- X-Authentication-Warning: fafnir.oit.pdx.edu: Host host-193-7.dhcp.pdx.edu [131.252.193.7] claimed to be [131.252.193.7] 6, 23 -- Message-ID: {44636912.8020908-at-pdx.edu} 6, 23 -- Date: Thu, 11 May 2006 09:40:50 -0700 6, 23 -- From: Rick Hugo {hugo-at-pdx.edu} 6, 23 -- Organization: Portland State University 6, 23 -- User-Agent: Thunderbird 1.5.0.2 (Windows/20060308) 6, 23 -- MIME-Version: 1.0 6, 23 -- To: gvrdolja-at-nature.berkeley.edu, 6, 23 -- Microscopy List {Microscopy-at-MSA.Microscopy.Com} 6, 23 -- Subject: Re: [Microscopy] cpd pains 6, 23 -- References: {200605102307.k4AN7rwa004554-at-ns.microscopy.com} 6, 23 -- In-Reply-To: {200605102307.k4AN7rwa004554-at-ns.microscopy.com} 6, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
All this talk of forcing needle valves and pliers, etc. on CPD systems is scaring me. There used to be a homemade CPD in the Berkeley Microlab which sounds very similar to the unit currently being discussed. It was retired in favor of a Tousimis unit chiefly because of safety concerns. The pressures and explosive-release volumes on these systems are sufficient to cause serious injury in the event of a failed valve or fitting. Trust me - It's Not Worth It. If you find you are using hand tools to adjust needle valves on these systems, you DO have a safety problem.
-------- Original Message --------
Right, it is HKL+Oxford now. Thanks for the correction.
TSL uses a Digiview 1612 Firewire camera that does a good job.
Depending on the grain size you are examining, probe diameter will be critical. Too large and small grains will be missed. Probe current increases frames per second--which is good.
TSL will do multi-phase EBSD. Their expansion of this is their Delphi option. I do not have this. I figure it is more for those trying to discover new things than sorting out what is basically known but not quantified. The plain OIM system comes with the TSL database and accepts the AMCS database....a very huge set of materials indeed.
EDAX EDS has drift correction (maps). This is the same basic shell used by TSL. Without correction, long scans at high mag are IMO useless and impossible. These are easy to spot since the resulting scans are wavy rather than consistent. Coating with C pretty much eliminates that element from analysis and also reduces signal by about 20-30%. High Z coating is not in the cards--too much absorption to produce diffraction patterns. VP is workable. Since one is not doing EDS maps, drift correction during EBSD collection is the key. EDAX/TSL also offers off-line dataset analysis for EBSD like they do for EDS. I'm not sure if HKL has the same option. I would ask about this when shopping.
gary g.
At 10:11 AM 5/11/2006, you wrote: } Gary, } } We're also considering a new EBSD system for an old LaB6 JEOL 840 that } delivers lots of beam current. It's to replace an old TSL system with a } deteriorated SIT camera. We're interested in the phase ID capabilities as } well as grain orientation analysis. } } Your comments on drift correction are especially interesting. I don't know } that a PGT/HKL connection exists any more. (Is PGT still in business?) HKL } is now part of Oxford, and they have at least partially integrated the EBSD } operation with Oxford's EDS software (INCA). INCA does has drift } correction, but I don't believe the combined system does yet. } } Larry } } } Larry Thomas } Pacific Northwest National Laboratory } Richland, WA 99352 } } email: Larry.Thomas-at-pnl.gov } phone: 509 372-0793 } -- } } } } } On 5/11/06 9:23 AM, "gary-at-gaugler.com" {gary-at-gaugler.com} wrote: } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } Sending to list--direct msgs bounce. If you } } have a better address, please advise and I will } } send any other material off-list. } } } } ------- } } } } What SEM do you plan on using? What type of } } gun does it use? } } } } What feature sizes are you interested in resolving? } } Ta and Cu seed layers are not possible to resolve } } even at .01u step size. I think that this is because } } no discernable grains have formed as yet. The electrodep Cu for } } damascene interconnects works well. } } } } I've done single crystal Sapphire, Silicon and micro } } crystal Si. What would your nano crystals be made of? } } If the material is non-conductive, this may pose a } } charging issue. Since EBSD penetrates only about 50nm, } } this does not leave much depth for coating. The } } preferred coating is C. However, I rarely coat any } } insulating material even at 20KV. } } } } There are basically two EBSD choices. TSL/EDAX or } } HKL/PGT. TSL and EDAX have been integrated for much } } longer than HKL and PGT (I think HKL connected just this } } year). TSL's camera/phosphor is round while HKL's camera end } } is square. This allows the HKL camera screen to get } } closer to the specimen. But I think that the TSL } } software complement is much better. AFAIK, only TSL } } has drift correction during data collection. At high } } mag and small step size, this is critical. However TSL's } } drift correction is not always repeatable and its own } } set of problems. But at least it is there and does } } usually work. } } } } The SEM is going to be at issue too since higher frames } } per second need more probe current. However, this is } } at the expense of probe diameter and lattice resolution. } } } } I'm using EDAX/TSL Pegasus with EDAX Genesis EDS and like it. } } The SEM is a Zeiss Supra 55VP, which has its own set } } of issues. } } } } gary g. } } } } } } At 03:34 AM 5/10/2006, you wrote: } } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } ---------------------------------------------------------------------------- } } } } } } Hi, } } } } } } We are considering buying EDX system integrated with EBSD for } } } elemental and "structural" analysis of inorganic materials in SEM. I hope } } } therefore that vendors of such equipment will contact me off the list. } } } However, I would be grateful also for any comments from listers on the } } } possibilities of EBSD as the method of structure identification of } } } individual nanocrystals in composite materials. } } } } } } Thank you, } } } } } } } } } } } } Leszek } } } } } } } } } } } } } } } } } } Leszek Kepinski } } } Institute of Low Temperature and Structure Research, } } } Polish Academy of Sciences, } } } P.O.Box 1410, } } } 50-950 Wroclaw, Poland } } } e-mail: L.Kepinski-at-int.pan.wroc.pl } } } } } } } } } } } } ==============================Original } Headers============================== } } } 13, 33 -- From L.Kepinski-at-int.pan.wroc.pl Wed May 10 05:31:44 2006 } } } 13, 33 -- Received: from mserv2.int.pan.wroc.pl } } } (mserv2.int.pan.wroc.pl [156.17.85.6]) } } } 13, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } } ESMTP id k4AAVh7k009241 } } } 13, 33 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 } } } 05:31:43 -0500 } } } 13, 33 -- Received: from localhost (localhost.localdomain [127.0.0.1]) } } } 13, 33 -- by mserv2.int.pan.wroc.pl (INTiBS (1.0)) with ESMTP } } } id 7143C134196 } } } 13, 33 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 } } } 12:43:45 +0200 (CEST) } } } 13, 33 -- Received: from mserv2.int.pan.wroc.pl ([127.0.0.1]) } } } 13, 33 -- by localhost (mserv2.int.pan.wroc.pl [127.0.0.1]) } } } (amavisd-new, port 10025) } } } 13, 33 -- with LMTP id 06540-02-6 for {Microscopy-at-microscopy.com} ; } } } 13, 33 -- Wed, 10 May 2006 12:43:43 +0200 (CEST) } } } 13, 33 -- X-Virus-Scanner: This message was checked by NOD32 } Antivirus system } } } 13, 33 -- NOD32 for Linux Mail Server. } } } 13, 33 -- For more information on NOD32 Antivirus System, } } } 13, 33 -- please, visit our website: http://www.nod32.com/. } } } 13, 33 -- Received: from b8p020 (sqd.int.pan.wroc.pl [156.17.85.20]) } } } 13, 33 -- by mserv2.int.pan.wroc.pl (INTiBS (1.0)) with SMTP } } } id 2ED7B134276 } } } 13, 33 -- for {Microscopy-at-microscopy.com} ; Wed, 10 May 2006 } } } 12:36:15 +0200 (CEST) } } } 13, 33 -- Message-ID: {006101c6741b$d020fe40$7f01a8c0-at-b8p020} } } } 13, 33 -- From: =?iso-8859-2?B?TC4gS+pwafFza2k=?= } } } {L.Kepinski-at-int.pan.wroc.pl} } } } 13, 33 -- To: {Microscopy-at-microscopy.com} } } } 13, 33 -- Subject: EDS_EBSD system } } } 13, 33 -- Date: Wed, 10 May 2006 12:23:43 +0200 } } } 13, 33 -- MIME-Version: 1.0 } } } 13, 33 -- Content-Type: text/plain; } } } 13, 33 -- charset="iso-8859-2" } } } 13, 33 -- Content-Transfer-Encoding: 7bit } } } 13, 33 -- X-Priority: 3 } } } 13, 33 -- X-MSMail-Priority: Normal } } } 13, 33 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 } } } 13, 33 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 } } } 13, 33 -- X-Scan-Module: SMTP[2006.04.28 (2006.01.25)] } } } 13, 33 -- X-Virus-Scanned: by amavisd-new at int.pan.wroc.pl } } } ==============================End of - } Headers============================== } } } } } } ==============================Original } Headers============================== } } 15, 19 -- From gary-at-gaugler.com Thu May 11 11:17:49 2006 } } 15, 19 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net } } [66.60.130.145]) } } 15, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } } k4BGHmMP004357 } } 15, 19 -- for {microscopy-at-microscopy.com} ; Thu, 11 May 2006 11:17:48 -0500 } } 15, 19 -- Received: (qmail 26316 invoked from network); 11 May } 2006 09:17:46 } } -0700 } } 15, 19 -- Received: by simscan 1.1.0 ppid: 26311, pid: 26312, t: 0.1909s } } 15, 19 -- scanners: regex: 1.1.0 attach: 1.1.0 } } 15, 19 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } } 15, 19 -- by qsmtp3 with SMTP; 11 May 2006 09:17:46 -0700 } } 15, 19 -- Message-Id: {7.0.1.0.2.20060511090631.02476f70-at-gaugler.com} } } 15, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } } 15, 19 -- Date: Thu, 11 May 2006 09:17:46 -0700 } } 15, 19 -- To: MSA listserver {microscopy-at-microscopy.com} } } 15, 19 -- From: Gary Gaugler {gary-at-gaugler.com} } } 15, 19 -- Subject: Re: [Microscopy] EDS_EBSD system } } 15, 19 -- In-Reply-To: {200605101034.k4AAYo4n012418-at-ns.microscopy.com} } } 15, 19 -- References: {200605101034.k4AAYo4n012418-at-ns.microscopy.com} } } 15, 19 -- Mime-Version: 1.0 } } 15, 19 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } } ==============================End of - } Headers==============================
==============================Original Headers============================== 10, 21 -- From gary-at-gaugler.com Thu May 11 13:23:11 2006 10, 21 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4BINAVK003464 10, 21 -- for {microscopy-at-microscopy.com} ; Thu, 11 May 2006 13:23:10 -0500 10, 21 -- Received: (qmail 3963 invoked from network); 11 May 2006 11:23:10 -0700 10, 21 -- Received: by simscan 1.1.0 ppid: 3960, pid: 3961, t: 0.1827s 10, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 21 -- by qsmtp1 with SMTP; 11 May 2006 11:23:09 -0700 10, 21 -- Message-Id: {7.0.1.0.2.20060511110407.02424110-at-gaugler.com} 10, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 21 -- Date: Thu, 11 May 2006 11:23:09 -0700 10, 21 -- To: "Thomas, Larry (PNNL)" {Larry.Thomas-at-pnl.gov} 10, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 21 -- Subject: Re: [Microscopy] Re: EDS_EBSD system 10, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 21 -- In-Reply-To: {C088BE6F.7E5%Larry.Thomas-at-pnl.gov} 10, 21 -- References: {200605111623.k4BGN9cK010183-at-ns.microscopy.com} 10, 21 -- {C088BE6F.7E5%Larry.Thomas-at-pnl.gov} 10, 21 -- Mime-Version: 1.0 10, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
The first question I would ask: WHY are the knobs hard to turn?
Generally it's a couple reasons.
#1 If the threads are lubricated and the grease/oil and enough of the VOCs (even if they aren't very volatile) dissipate the lubricant can turn to glue. -Solution: Disassemble, clean (with WD-40, CRC or other solvating lubricants), and re-apply some lightweight grease (lithium or wheel bearing would be fine), or a touch of heavy oil.
#2 If the threads/valve are damaged, you probably should replace the valve.
These all are extremely simple devices. And as such parts are relatively easy to repair/replace or fix. Assuming you have the patience to source the proper valves. The pressures the CPDs the microscopy community utilizes are relatively insignificant to some of the devices folks in Physics and engineering play with daily, not to mention, McMaster-Carr has a great selection of valves and what not that are rated well above the typical 1500-3000 psi burst limit (safety) on the CPDs.
If the valve was always hard to turn that's one thing, if it is now, make it easy to turn.
Try even spraying the threads with some WD-40 or liquid Wrench and run a cycle or two and see if that helps anything.
I am a big fan of the Polaron type CPD. I've never been let down by one, and it is the best model for 'teaching' the principles... no black boxes and although manual, no sane minded individual should start a CPD run and let it go un-attended... so why not be manual? My apologies to those who sell automatic ones, this is just my opinion.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
Sent: Wednesday, May 10, 2006 7:10 PM To: Williams, Geoffrey
Hello, I'm using an older Pelco CPD 2 critical point drier. Anyone use this? It has these gnurled steel knobs for opening/closing the valves. Controlling the valves kills your fingers since the rough metal is hard on your skin. Anyone modified it with plastic knobs or rubber covers or something to make it less of a literal pain to do a critical point drying run?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \/\/\ 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============================== 19, 29 -- From Geoffrey_Williams-at-brown.edu Thu May 11 14:29:30 2006 19, 29 -- Received: from pyxis.services.brown.edu (pyxis.services.brown.edu [128.148.106.154]) 19, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4BJTUwW014301 19, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 11 May 2006 14:29:30 -0500 19, 29 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 19, 29 -- by pyxis.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k4BJTT1G029302 19, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 11 May 2006 15:29:30 -0400 (EDT) 19, 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); 19, 29 -- Thu, 11 May 2006 15:29:29 -0400 19, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 19, 29 -- Content-class: urn:content-classes:message 19, 29 -- MIME-Version: 1.0 19, 29 -- Content-Type: text/plain; 19, 29 -- charset="US-ASCII" 19, 29 -- Subject: RE: cpd pains 19, 29 -- Date: Thu, 11 May 2006 15:29:27 -0400 19, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F044FFAB6-at-MAIL1.AD.Brown.Edu} 19, 29 -- X-MS-Has-Attach: 19, 29 -- X-MS-TNEF-Correlator: 19, 29 -- Thread-Topic: cpd pains 19, 29 -- Thread-Index: AcZ0hu6v536bZuTJRHefGkmGyiABXgAp6KHg 19, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 19, 29 -- To: {Microscopy-at-microscopy.com} 19, 29 -- X-OriginalArrivalTime: 11 May 2006 19:29:29.0855 (UTC) FILETIME=[391AE8F0:01C67531] 19, 29 -- X-Brown-Proofpoint: Not Infected 19, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06042601 definitions=3.0.0-06051106 19, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06042601 definitions=3.0.0-06051106 19, 29 -- Content-Transfer-Encoding: 8bit 19, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4BJTUwW014301 ==============================End of - Headers==============================
I am writing to get information on web-pages which have compiled links to bio-imaging and microscopy facilities both within the US and internationally.
I have searched and located a few such web-sites, but they are not very comprehensive and many of the links are dead.
If anyone in the community may have web-addresses or information on sites of this nature, I would appreciate their input.
Thank you,
David Lowry School of Life Sciences Arizona State University Tempe, AZ 85287-4501 office: 480-727-0725 lab: 480-965-2463
==============================Original Headers============================== 10, 26 -- From dlowry-at-asu.edu Thu May 11 15:13:41 2006 10, 26 -- Received: from post5.inre.asu.edu (post5.inre.asu.edu [129.219.110.120]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4BKDeKs025039 10, 26 -- for {Microscopy-at-Microscopy.com} ; Thu, 11 May 2006 15:13:40 -0500 10, 26 -- Received: from conversion.post5.inre.asu.edu by asu.edu (PMDF V6.1-1X6 #30769) 10, 26 -- id {0IZ400A01AOYKN-at-asu.edu} for Microscopy-at-Microscopy.com; Thu, 10, 26 -- 11 May 2006 13:10:10 -0700 (MST) 10, 26 -- Received: from EX03.asurite.ad.asu.edu 10, 26 -- (excl1-a0.asurite.ad.asu.edu [129.219.12.199]) 10, 26 -- by asu.edu (PMDF V6.1-1X6 #30769) with ESMTP id {0IZ40094UAOXGF-at-asu.edu} for 10, 26 -- Microscopy-at-Microscopy.com; Thu, 11 May 2006 13:10:09 -0700 (MST) 10, 26 -- Date: Thu, 11 May 2006 13:10:10 -0700 10, 26 -- From: David Lowry {dlowry-at-asu.edu} 10, 26 -- Subject: 10, 26 -- To: Microscopy-at-Microscopy.com 10, 26 -- Cc: Douglas Chandler {d.chandler-at-asu.edu} , Robert Roberson {robby2-at-asu.edu} 10, 26 -- Message-id: {B9366E69DACF09458B402E4C4385012601188099-at-EX03.asurite.ad.asu.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 10, 26 -- Content-type: text/plain; charset=iso-8859-1 10, 26 -- Thread-Index: AcZ1NueqXPUlUZH9SzS8YvyHUcm0cw== 10, 26 -- Content-class: urn:content-classes:message 10, 26 -- X-MS-Has-Attach: 10, 26 -- X-MS-TNEF-Correlator: 10, 26 -- Content-Transfer-Encoding: 8bit 10, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4BKDeKs025039 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (cheetham.3-at-osu.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, May 11, 2006 at 16:42:15 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both cheetham.3-at-osu.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: cheetham.3-at-osu.edu Name: Sonia Cheetham
Organization: The Ohio State University
Education: Graduate College
Location: Wooster , Ohio
Title: question on photomicrophotography
Question: I have being trying to find out what photomicrophotography is about. I came across this term in a paper regarding confirmation of PMO getting into the cells. Can you give me a brief exlanation of what is this technique? Thanks
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Email: junhe-at-unmc.edu Name: Jun
Organization: unmc
Title-Subject: [Filtered] Freon113 in Dehydration and Critical Point Drying
Question: I have a question on the role of Froen 113 in the common route of: wet specimen---70% to 100% Ethanol----Freon113--Liduid CO2
CO2 is the common transitional fluid. Ethanol is the dehydration fluid. I know that in some cases people go from Ethanol to CO2 directely. But, Since the above route is also mentioned, I wonder what advantage Froen113 brings to the preparation process.
Ann Fook Yang EM Unit/ Unite EM AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 yanga-at-agr.gc.ca Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: dlowry-at-asu.edu [mailto:dlowry-at-asu.edu] Sent: Thursday, May 11, 2006 4:18 PM To: Yang, Ann-Fook
Colleagues,
I am writing to get information on web-pages which have compiled links to bio-imaging and microscopy facilities both within the US and internationally.
I have searched and located a few such web-sites, but they are not very comprehensive and many of the links are dead.
If anyone in the community may have web-addresses or information on sites of this nature, I would appreciate their input.
Thank you,
David Lowry School of Life Sciences Arizona State University Tempe, AZ 85287-4501 office: 480-727-0725 lab: 480-965-2463
==============================Original Headers============================== 10, 26 -- From dlowry-at-asu.edu Thu May 11 15:13:41 2006 10, 26 -- Received: from post5.inre.asu.edu (post5.inre.asu.edu [129.219.110.120]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4BKDeKs025039 10, 26 -- for {Microscopy-at-Microscopy.com} ; Thu, 11 May 2006 15:13:40 -0500 10, 26 -- Received: from conversion.post5.inre.asu.edu by asu.edu (PMDF V6.1-1X6 #30769) 10, 26 -- id {0IZ400A01AOYKN-at-asu.edu} for Microscopy-at-Microscopy.com; Thu, 10, 26 -- 11 May 2006 13:10:10 -0700 (MST) 10, 26 -- Received: from EX03.asurite.ad.asu.edu 10, 26 -- (excl1-a0.asurite.ad.asu.edu [129.219.12.199]) 10, 26 -- by asu.edu (PMDF V6.1-1X6 #30769) with ESMTP id {0IZ40094UAOXGF-at-asu.edu} for 10, 26 -- Microscopy-at-Microscopy.com; Thu, 11 May 2006 13:10:09 -0700 (MST) 10, 26 -- Date: Thu, 11 May 2006 13:10:10 -0700 10, 26 -- From: David Lowry {dlowry-at-asu.edu} 10, 26 -- Subject: 10, 26 -- To: Microscopy-at-Microscopy.com 10, 26 -- Cc: Douglas Chandler {d.chandler-at-asu.edu} , Robert Roberson {robby2-at-asu.edu} 10, 26 -- Message-id: {B9366E69DACF09458B402E4C4385012601188099-at-EX03.asurite.ad.asu.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 10, 26 -- Content-type: text/plain; charset=iso-8859-1 10, 26 -- Thread-Index: AcZ1NueqXPUlUZH9SzS8YvyHUcm0cw== 10, 26 -- Content-class: urn:content-classes:message 10, 26 -- X-MS-Has-Attach: 10, 26 -- X-MS-TNEF-Correlator: 10, 26 -- Content-Transfer-Encoding: 8bit 10, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4BKDeKs025039 ==============================End of - Headers==============================
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We're trying to investigate bacteria biofilms on a latex material (like surgical gloves) with TEM. We couldn't cut the latex material with our regular glass knives. Is there any suggestion about this problem?
Thanks for any comment...
Dr. Necat Yilmaz Mersin University Medical School Histology & Embryolgy Dept.
We are looking for a lab to do some contract work, embedding, sectioning and H&E staining of zebrafish using JB4/GMA plastics. We will provide fixed specimens. Ultimately we would like serial sections but for now we just want to see what sort of results we can expect. Please contact me directly. Nothe that I will be out of the office next week (15th-19th) so my response may be delayed. Thanks.
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 6, 33 -- From mcauliff-at-umdnj.edu Fri May 12 12:40:53 2006 6, 33 -- Received: from zix02.umdnj.edu (zix02.UMDNJ.EDU [130.219.34.125]) 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4CHeq10031341 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Fri, 12 May 2006 12:40:52 -0500 6, 33 -- Received: from zix02.umdnj.edu (ZixVPM [127.0.0.1]) 6, 33 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 9E1464BE1E 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Fri, 12 May 2006 12:40:51 -0500 (CDT) 6, 33 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 6, 33 -- by zix02.umdnj.edu (Proprietary) with ESMTP id C8ECC4BDAD 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Fri, 12 May 2006 12:40:49 -0500 (CDT) 6, 33 -- Received: from ([130.219.34.131]) 6, 33 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.13635540; 6, 33 -- Fri, 12 May 2006 13:40:20 -0400 6, 33 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 33 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 33 -- id {0IZ500F01XNDSY-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 33 -- for microscopy-at-msa.microscopy.com; Fri, 12 May 2006 13:40:20 -0400 (EDT) 6, 33 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 33 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 33 -- 2004)) with ESMTP id {0IZ5006KPYF739-at-Polaris.umdnj.edu} ; Fri, 6, 33 -- 12 May 2006 13:40:20 -0400 (EDT) 6, 33 -- Date: Fri, 12 May 2006 13:41:17 -0400 6, 33 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 33 -- Subject: GMA/JB4 Contract work 6, 33 -- To: Histonet {histonet-at-pathology.swmed.edu} , 6, 33 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 6, 33 -- Message-id: {4464C8BD.7090000-at-umdnj.edu} 6, 33 -- MIME-version: 1.0 6, 33 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 33 -- Content-transfer-encoding: 7BIT 6, 33 -- X-Accept-Language: en-us, en 6, 33 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 33 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
At the risk of putting my foot in my oversized mouth - - - You need a cryo-microtome and work around -40. This gets you below the glass transistion temperature for most elastomers and the latex will no longer behave like an elastic material, but a hard brittle glass. You may need to fiddle with temperature and pick-up technique. I've sectioned polymer and and used both glycerin/water, mineral spirits/xylene and DMSO/water depending on the temperature and my end goal (Light, SEM or TEM). A diamond knife and boat would be my prefered method. I like to pick up with a "perfect loop" and place on carbon coated grid.
good luck and have fun........
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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nyilmaz-at-mersin.ed u.tr To: frank.karl-at-degussa.com cc: 05/12/2006 08:44 Subject: [Microscopy] viaWWW: Cutting latex for TEM AM Please respond to nyilmaz
Title-Subject: [Filtered] Cutting latex for TEM
Question: Hello Everybody...
We're trying to investigate bacteria biofilms on a latex material (like surgical gloves) with TEM. We couldn't cut the latex material with our regular glass knives. Is there any suggestion about this problem?
Thanks for any comment...
Dr. Necat Yilmaz Mersin University Medical School Histology & Embryolgy Dept.
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 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] Locust brain
Question: Hello ListServers,
Does anyone have a good reference or two for images of the locust brain? It would be great if I could find both LM and TEM images.
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Email: dgmorgan-at-ucdavis.edu Name: David Morgan
Organization: University of California, Davis
Title-Subject: [Filtered] polymer microtomy
Question: I am forwarding two different questions regarding the microtomy of polymers from associates, and if additional information would be useful, please let me know and I will find out what else I can.
1) polymer embedded ZnO nano-rods. I do not know what type of polymer this specimen contains, which is probably critical to any advice that might be offered. I suspect that we will need to use a cryo-microtome and simply experiment to determine the best temperature to use. Any and all suggestions would be most welcome. Also, does anyone have experience with ZnO nano-rods, especially with regard to whether this material is capable of damaging a diamond knife?
2) poly-styrene beads. Another colleague is potentially interested in sectioning ~6 micron beads. Suggestions about the best way to handle such a sample would be welcome - the proper embedding material, sectioning temperature, etc.
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 AMCGroup2-at-aol.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: AMCGroup2-at-aol.com Name: James Glossinger
Organization: AMC Group
Title-Subject: TEM Contractor
We are currently seeking an independent TEM analyst, academic/non-profit TEM facility or, commercial TEM lab for contracting ongoing advanced TEM imaging and AEM projects, involving semiconductor materials and devices.
If interested and qualified, please contact me off-line via email.
Thanks, Jim
--------------------- James Glossinger, Ph.D. Principal Scientist AMC Group
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Email: palladineus-at-yahoo.com Name: Randy Khoo
Title-Subject: [Filtered] Masson-Fontana staining for melanin
Question: I am wondering if anybody can help me out on a poser that I have.
Melanin is able to reduce the ammoniacal silver nitrate solution without the addition of an external reducing agent. However, as I understand it, melanin exists in 2 forms, both in oxidized form and reduced form. So, is the reduced form only responsible for reducing the silver nitrate? If so, can the addition of a reducing agent result in both forms reducing the silver nitrate and show a better staining profile? Can this method be used to semi-quantify both the reduced and oxidized forms?
I have a project where I need to positively identify human cells that may have mouse cells mixed with them in a tumor. Does anyone have a suggestion for a protocol that will positively identify a cell as mouse or as human. Antibodies that I have tried so far have not worked, post-embedding.
Thanks, Greg -- Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 2, 23 -- From gwe-at-ufl.edu Mon May 15 08:18:13 2006 2, 23 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 2, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FDID2j018130 2, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 15 May 2006 08:18:13 -0500 2, 23 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 2, 23 -- (authenticated bits=0) 2, 23 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k4FDIB1j1122394 2, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 15 May 2006 09:18:12 -0400 2, 23 -- Message-ID: {44687F92.8060003-at-ufl.edu} 2, 23 -- Date: Mon, 15 May 2006 09:18:10 -0400 2, 23 -- From: greg {gwe-at-ufl.edu} 2, 23 -- Reply-To: gwe-at-ufl.edu 2, 23 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 2, 23 -- X-Accept-Language: en-us, en 2, 23 -- MIME-Version: 1.0 2, 23 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 2, 23 -- Subject: Mouse -vs-Human 2, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 23 -- Content-Transfer-Encoding: 7bit 2, 23 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 2, 23 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 2, 23 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 2, 23 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Ah... I'm disappointed. I thought we were going to have a nice discussion about the transition from human adjustments (knobs) to using the mouse (bar sliders)....
I was already to pull out the link to http://www.griffintechnology.com/products/powermate/ ... suggesting that maybe it is the answer we old school knob aficionados have been waiting for.
Alas I have no suggestions relating to the topic. So please accept or excuse this mild attempt at being funny on a Monday morning.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: gwe-at-ufl.edu [mailto:gwe-at-ufl.edu] Sent: Monday, May 15, 2006 9:22 AM To: Williams, Geoffrey
I have a project where I need to positively identify human cells that may have mouse cells mixed with them in a tumor. Does anyone have a suggestion for a protocol that will positively identify a cell as mouse or as human. Antibodies that I have tried so far have not worked, post-embedding.
Thanks, Greg -- Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Director, EM Core Lab P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu Phone: 352-392-1295 Fax: 352-846-0251
==============================Original Headers============================== 2, 23 -- From gwe-at-ufl.edu Mon May 15 08:18:13 2006 2, 23 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 2, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FDID2j018130 2, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 15 May 2006 08:18:13 -0500 2, 23 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 2, 23 -- (authenticated bits=0) 2, 23 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with ESMTP id k4FDIB1j1122394 2, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 15 May 2006 09:18:12 -0400 2, 23 -- Message-ID: {44687F92.8060003-at-ufl.edu} 2, 23 -- Date: Mon, 15 May 2006 09:18:10 -0400 2, 23 -- From: greg {gwe-at-ufl.edu} 2, 23 -- Reply-To: gwe-at-ufl.edu 2, 23 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 2, 23 -- X-Accept-Language: en-us, en 2, 23 -- MIME-Version: 1.0 2, 23 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 2, 23 -- Subject: Mouse -vs-Human 2, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 23 -- Content-Transfer-Encoding: 7bit 2, 23 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 2, 23 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 2, 23 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 2, 23 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
==============================Original Headers============================== 12, 29 -- From Geoffrey_Williams-at-brown.edu Mon May 15 08:29:50 2006 12, 29 -- Received: from pyxis.services.brown.edu (pyxis.services.brown.edu [128.148.106.154]) 12, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FDToxx028190 12, 29 -- for {Microscopy-at-microscopy.com} ; Mon, 15 May 2006 08:29:50 -0500 12, 29 -- Received: from ex-gateway1-out.AD.Brown.Edu (ex-gateway1.ad.brown.edu [128.148.21.51]) 12, 29 -- by pyxis.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k4FDTdO1011626; 12, 29 -- Mon, 15 May 2006 09:29:49 -0400 (EDT) 12, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.31]) by ex-gateway1-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 12, 29 -- Mon, 15 May 2006 09:29:47 -0400 12, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 12, 29 -- Content-class: urn:content-classes:message 12, 29 -- MIME-Version: 1.0 12, 29 -- Content-Type: text/plain; 12, 29 -- charset="US-ASCII" 12, 29 -- Subject: RE: [Microscopy] Mouse -vs-Human 12, 29 -- Date: Mon, 15 May 2006 09:29:47 -0400 12, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F044FFC0C-at-MAIL1.AD.Brown.Edu} 12, 29 -- X-MS-Has-Attach: 12, 29 -- X-MS-TNEF-Correlator: 12, 29 -- Thread-Topic: [Microscopy] Mouse -vs-Human 12, 29 -- Thread-Index: AcZ4IoOHi5Mh71SqR4y2kC4rjGtdoAAALxag 12, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 12, 29 -- To: {gwe-at-ufl.edu} , {Microscopy-at-microscopy.com} 12, 29 -- X-OriginalArrivalTime: 15 May 2006 13:29:47.0713 (UTC) FILETIME=[A2CC6310:01C67823] 12, 29 -- X-Brown-Proofpoint: Not Infected 12, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06042601 definitions=3.0.0-06051209 12, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06042601 definitions=3.0.0-06051209 12, 29 -- Content-Transfer-Encoding: 8bit 12, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4FDToxx028190 ==============================End of - Headers==============================
Does anyone where to find one of the spatial calibration slides that used to be sold by a company called Richardson Technologies? The company appears to be gone but I'm trying to get hold of one the slides.
Thank you!
--David Hitrys QImaging Corporation
==============================Original Headers============================== 1, 20 -- From dhitrys-at-qimaging.com Mon May 15 08:39:54 2006 1, 20 -- Received: from smtp01.lnh.mail.rcn.net (smtp01.lnh.mail.rcn.net [207.172.4.11]) 1, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FDdsFA005775 1, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 15 May 2006 08:39:54 -0500 1, 20 -- Received: from 209-6-80-225.c3-0.frm-ubr2.sbo-frm.ma.cable.rcn.com (HELO DHPC) ([209.6.80.225]) 1, 20 -- by smtp01.lnh.mail.rcn.net with ESMTP; 15 May 2006 09:40:40 -0400 1, 20 -- X-IronPort-AV: i="4.05,130,1146456000"; 1, 20 -- d="scan'208"; a="203995933:sNHT1448836834" 1, 20 -- From: "David Hitrys" {dhitrys-at-qimaging.com} 1, 20 -- To: {Microscopy-at-microscopy.com} 1, 20 -- Subject: Spatial Calibration Slide from Richardson Technologies 1, 20 -- Date: Mon, 15 May 2006 09:39:39 -0400 1, 20 -- Message-ID: {003401c67825$03e01ce0$0302a8c0-at-DHPC} 1, 20 -- MIME-Version: 1.0 1, 20 -- Content-Type: text/plain; 1, 20 -- charset="us-ascii" 1, 20 -- Content-Transfer-Encoding: 7bit 1, 20 -- X-Mailer: Microsoft Office Outlook 11 1, 20 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 1, 20 -- Thread-index: AcZ4JQOdwz3Z0tRTQpK666aP0DxGaQ== ==============================End of - Headers==============================
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 08:42 AM 5/15/2006, dhitrys-at-qimaging.com wrote:
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==============================Original Headers============================== 14, 18 -- From bfoster-at-mme1.com Mon May 15 11:08:05 2006 14, 18 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 14, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FG85cl022948 14, 18 -- for {microscopy-at-microscopy.com} ; Mon, 15 May 2006 11:08:05 -0500 14, 18 -- Received: (qmail 12441 invoked by uid 2020); 15 May 2006 11:24:05 -0500 14, 18 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 14, 18 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 15 May 2006 11:24:05 -0500 14, 18 -- Message-Id: {7.0.1.0.0.20060515110734.02067758-at-mme1.com} 14, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 18 -- Date: Mon, 15 May 2006 11:08:15 -0500 14, 18 -- To: dhitrys-at-qimaging.com, microscopy Listserver {microscopy-at-microscopy.com} 14, 18 -- From: Barbara Foster {bfoster-at-mme1.com} 14, 18 -- Subject: Re: [Microscopy] Spatial Calibration Slide from Richardson 14, 18 -- Technologies 14, 18 -- In-Reply-To: {200605151342.k4FDgKaa009921-at-ns.microscopy.com} 14, 18 -- References: {200605151342.k4FDgKaa009921-at-ns.microscopy.com} 14, 18 -- Mime-Version: 1.0 14, 18 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
In response to a recent question on the ListServer which, in part, asked about the status of PGT Microanalysis, I would like to reply:
On Nov 17, 2005 Bruker AXS announced the acquisition of Princeton Gamma-Tech Microanalysis and Röntec AG. These two organizations were merged to form Bruker AXS Microanalysis. Bruker AXS develops and manufactures a broad range of analytical X-ray systems including XRF, XRD and now, X-ray Microanalysis.
Customer service and support for both PGT Microanalysis and Röntec have been expanded as part of the Bruker AXS worldwide operation. PGT and Röntec customers should contact Bruker AXS for continued product and applications support.
Bruker AXS Microanalysis is a proud Sustaining Member of MSA and MAS, carrying on the long time support given by PGT Microanalysis and Röntec to these fine professional organizations.
Doug Skinner Assistant Vice President Bruker-AXS Microanalysis 609-771-4400 Doug.Skinner-at-Bruker-AXS.com www.bruker-axs-ma.com
==============================Original Headers============================== 10, 23 -- From Doug.Skinner-at-bruker-axs.com Mon May 15 12:34:01 2006 10, 23 -- Received: from isa1.bruker-axs.com (68-22-68-158.ded.ameritech.net [68.22.68.158]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FHY0Gf001851 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 15 May 2006 12:34:01 -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 -- Mon, 15 May 2006 12:34:42 -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="iso-8859-1" 10, 23 -- Subject: Status of Princeton Gamma-Tech Microanalysis (PGT) 10, 23 -- Date: Mon, 15 May 2006 12:34:42 -0500 10, 23 -- Message-ID: {235D0F9F0400B3449E8C229D9D24CC57024C2EE3-at-mail1.bruker-axs.com} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Status of Princeton Gamma-Tech Microanalysis (PGT) 10, 23 -- Thread-Index: AcZ4RdBgdQwFFMgTR4CD5qIjFruOCw== 10, 23 -- From: "Skinner, Doug" {Doug.Skinner-at-bruker-axs.com} 10, 23 -- To: {Microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 15 May 2006 17:34:42.0631 (UTC) FILETIME=[D9A6ED70:01C67845] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4FHY0Gf001851 ==============================End of - Headers==============================
Does anybody have any experience with this digital microscope camera? I don't see on the webpage I found that the chip is cooled. How opinions about the image quality? Reliability? The price is attractive. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Mon May 15 16:14:10 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FLE9wk016011 3, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 15 May 2006 16:14:09 -0500 3, 20 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k4FLo84l006919 3, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 15 May 2006 17:50:08 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 3, 20 -- Mon, 15 May 2006 17:14:05 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f06230916c08e9f8899eb-at-[141.209.160.132]} 3, 20 -- Date: Mon, 15 May 2006 17:14:07 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: Polaroid DMC2 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 15 May 2006 21:14:05.0814 (UTC) FILETIME=[7F854160:01C67864] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -4 () L_EXCH_MF 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================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 lewryan-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: lewryan-at-gmail.com Name: Ryan
Organization: Dalhousie University
Title-Subject: [Filtered] SEM imaging on sample with magnetic substrate
Question: I have a saple with deposits of an alloy of Sn-Co-C on a magnetic stainless steel substrate (430 stainless steel). I'm using a cold field emission microscope and am having trouble getting high resolution images. Could you suggest what setting I should use to get started?
(honest, that's the page URL, I got it from Googling polaroid dmc2)
is says that it's no longer manufactured.
seems a pity
cheers
rtch
On 15 May 2006 at 16:16, oshel1pe-at-cmich.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 } ---------------------------------------------------------------------------- } } Listers, } } Does anybody have any experience with this digital microscope camera? } I don't see on the webpage I found that the chip is cooled. } How opinions about the image quality? Reliability? The price is attractive. } Thanks. } } Phil } -- } Philip Oshel } Microscopy Facility Supervisor } Department of Biology } Central Michigan University } 024C Brooks Hall } Mt. Pleasant, MI 48859 } (989) 774-3576 }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 16, 27 -- From r.sims-at-auckland.ac.nz Mon May 15 17:29:41 2006 16, 27 -- Received: from zeppo.itss.auckland.ac.nz (zeppo.itss.auckland.ac.nz [130.216.190.14]) 16, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FMTeeX004396 16, 27 -- for {Microscopy-at-microscopy.com} ; Mon, 15 May 2006 17:29:41 -0500 16, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 16, 27 -- by zeppo.itss.auckland.ac.nz (Postfix) with ESMTP id CF93A3546E; 16, 27 -- Tue, 16 May 2006 10:29:39 +1200 (NZST) 16, 27 -- Received: from zeppo.itss.auckland.ac.nz ([127.0.0.1]) 16, 27 -- by localhost (smtpd.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 16, 27 -- with ESMTP id 21284-30; Tue, 16 May 2006 10:29:39 +1200 (NZST) 16, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 16, 27 -- by zeppo.itss.auckland.ac.nz (Postfix) with ESMTP id B15BD353A5; 16, 27 -- Tue, 16 May 2006 10:29:39 +1200 (NZST) 16, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 16, 27 -- To: oshel1pe-at-cmich.edu 16, 27 -- Date: Tue, 16 May 2006 10:33:26 +1200 16, 27 -- MIME-Version: 1.0 16, 27 -- Subject: Re: [Microscopy] Polaroid DMC2 16, 27 -- Cc: Microscopy-at-microscopy.com 16, 27 -- Message-ID: {4469AA76.24160.7DE2DE-at-localhost} 16, 27 -- Priority: normal 16, 27 -- In-reply-to: {200605152116.k4FLGVBG017899-at-ns.microscopy.com} 16, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 16, 27 -- Content-type: text/plain; charset=US-ASCII 16, 27 -- Content-transfer-encoding: 7BIT 16, 27 -- Content-description: Mail message body 16, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
If your purpose is just to identify mouse cells in a human culture, you don't need EM. Actually it would be much easier in LM. Any way, you could think about using HLA marker, they are external (it's just an idea I have no experience with that).
regards,
Stephane
--- gwe-at-ufl.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a project where I need to positively identify } human cells that } may have mouse cells mixed with them in a tumor. } Does anyone have a } suggestion for a protocol that will positively } identify a cell as mouse } or as human. Antibodies that I have tried so far } have not worked, } post-embedding. } } Thanks, Greg } -- } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } Scientific Director, EM Core Lab } P.O. Box 118525 } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } Phone: 352-392-1295 } Fax: 352-846-0251 } } ==============================Original } Headers============================== } 2, 23 -- From gwe-at-ufl.edu Mon May 15 08:18:13 2006 } 2, 23 -- Received: from smtp.ufl.edu } (smtp02.osg.ufl.edu [128.227.74.165]) } 2, 23 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k4FDID2j018130 } 2, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, } 15 May 2006 08:18:13 -0500 } 2, 23 -- Received: from [10.227.60.63] } (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) } 2, 23 -- (authenticated bits=0) } 2, 23 -- by smtp.ufl.edu (8.13.6/8.13.6/2.5.1) with } ESMTP id k4FDIB1j1122394 } 2, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, } 15 May 2006 09:18:12 -0400 } 2, 23 -- Message-ID: {44687F92.8060003-at-ufl.edu} } 2, 23 -- Date: Mon, 15 May 2006 09:18:10 -0400 } 2, 23 -- From: greg {gwe-at-ufl.edu} } 2, 23 -- Reply-To: gwe-at-ufl.edu } 2, 23 -- User-Agent: Mozilla Thunderbird 1.0.7 } (Windows/20050923) } 2, 23 -- X-Accept-Language: en-us, en } 2, 23 -- MIME-Version: 1.0 } 2, 23 -- To: "Microscopy-at-sparc5.microscopy.com" } {Microscopy-at-MSA.Microscopy.Com} } 2, 23 -- Subject: Mouse -vs-Human } 2, 23 -- Content-Type: text/plain; } charset=ISO-8859-1; format=flowed } 2, 23 -- Content-Transfer-Encoding: 7bit } 2, 23 -- X-Spam-Status: hits=-1.44, required=5, } tests=ALL_TRUSTED } 2, 23 -- X-UFL-Spam-Status: hits=-1.44, required=5, } tests=ALL_TRUSTED } 2, 23 -- X-Scanned-By: CNS Open Systems Group } (http://open-systems.ufl.edu/services/smtp-relay/) } 2, 23 -- X-UFL-Scanned-By: CNS Open Systems Group } (http://open-systems.ufl.edu/services/smtp-relay/) } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Tue May 16 03:56:59 2006 9, 20 -- Received: from web37409.mail.mud.yahoo.com (web37409.mail.mud.yahoo.com [209.191.87.62]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4G8uxm1001022 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 03:56:59 -0500 9, 20 -- Received: (qmail 62761 invoked by uid 60001); 16 May 2006 08:56:58 -0000 9, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 20 -- s=s1024; d=yahoo.com; 9, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 20 -- b=vkjIObXA9f/aAyD94BBUxAzib2BM5EVypr+zAyrZbcJXw3DbTY9XHamLv+w8EwXUZoBveEPbQFuW/7B/ntgqlD4+vmHjziXn81Df70FmpPXvsfoVxU2WCHF7S12cpP3LehOC+8BkcXw+b1m1HCfeSsveM+1eEKv6JU7u7TmFRtc= ; 9, 20 -- Message-ID: {20060516085658.62759.qmail-at-web37409.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37409.mail.mud.yahoo.com via HTTP; Tue, 16 May 2006 01:56:58 PDT 9, 20 -- Date: Tue, 16 May 2006 01:56:58 -0700 (PDT) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] Mouse -vs-Human 9, 20 -- To: gwe-at-ufl.edu 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200605151322.k4FDMVbw027060-at-ns.microscopy.com} 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; charset=iso-8859-1 9, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
We are considering Fovea Pro as a set of quantitation plug-ins for Photoshop. Can I ask those who use FP, and who also teach IA, to contact me off list? I'd like to get an idea of the user base, and accumulate any thoughts on re-establishing an FP forum.
Several years ago many (most?) of the EM supply vendors switched the formula for carbon tabs to a "stiff", non-flexible formula. I was told at that time that the old formula used typewriter tape as a component.
These new stiff tabs are unsuitable for many of the preps we do. I was able to find the old formula at Fullam and now they have switched too.
Does anyone know where to find truly flexible carbon tabs. The ones Fullam had were from Nisshin EM, very expensive but quite high quality.
Thanks in advance.
Owen Mills MTU, Houghton MI
==============================Original Headers============================== 5, 31 -- From opmills-at-mtu.edu Tue May 16 07:48:57 2006 5, 31 -- Received: from node7.mtu.edu (node7.mtu.edu [141.219.69.7]) 5, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GCmvf9024068 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 07:48:57 -0500 5, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 5, 31 -- by node7.mtu.edu (8.13.1/8.13.1) with ESMTP id k4GCmu39024410 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:56 -0400 5, 31 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 5, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k4GCmuco022544 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:56 -0400 5, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) 5, 31 -- by node34.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k4GCmu6b023709 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:56 -0400 5, 31 -- (envelope-from opmills-at-mtu.edu) 5, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 5, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k4GCmtjx010574 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:55 -0400 (EDT) 5, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 5, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k4GCmtRx016397 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:55 -0400 5, 31 -- Mime-Version: 1.0 (Apple Message framework v749.3) 5, 31 -- Content-Transfer-Encoding: 7bit 5, 31 -- Message-Id: {E9327AEB-33E3-4E6C-9DA6-F2AA87734FF8-at-mtu.edu} 5, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 5, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 5, 31 -- Subject: dbl stick carbon tab blues 5, 31 -- Date: Tue, 16 May 2006 08:49:15 -0400 5, 31 -- X-Mailer: Apple Mail (2.749.3) 5, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.5.16.53106 5, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__CP_MEDIA_2_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
Have you looked at Ted Pella's assortment (Pelco)? Some answers to your questions may be found in his letter above. Best wishes,
Jim
-----Original Message----- X-from: opmills-at-mtu.edu [mailto:opmills-at-mtu.edu] Sent: Tuesday, May 16, 2006 2:56 PM To: James Chalcroft
Several years ago many (most?) of the EM supply vendors switched the formula for carbon tabs to a "stiff", non-flexible formula. I was told at that time that the old formula used typewriter tape as a component.
These new stiff tabs are unsuitable for many of the preps we do. I was able to find the old formula at Fullam and now they have switched too.
Does anyone know where to find truly flexible carbon tabs. The ones Fullam had were from Nisshin EM, very expensive but quite high quality.
Thanks in advance.
Owen Mills MTU, Houghton MI
==============================Original Headers============================== 16, 23 -- From jchalcro-at-neuro.mpg.de Tue May 16 08:26:15 2006 16, 23 -- Received: from neuro.mpg.de (mail.neuro.mpg.de [130.183.250.1]) 16, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GDQEre001944 16, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:26:15 -0500 16, 23 -- Content-class: urn:content-classes:message 16, 23 -- Return-Receipt-To: "James Chalcroft" {jchalcro-at-neuro.mpg.de} 16, 23 -- MIME-Version: 1.0 16, 23 -- Content-Type: text/plain; 16, 23 -- charset="US-ASCII" 16, 23 -- Disposition-Notification-To: "James Chalcroft" {jchalcro-at-neuro.mpg.de} 16, 23 -- Subject: RE: [Microscopy] dbl stick carbon tab blues 16, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 23 -- Date: Tue, 16 May 2006 15:26:13 +0200 16, 23 -- Message-ID: {6BBC089D4E1E87459875FB80D8030031BE21D2-at-s6.neuro.mpg.de} 16, 23 -- X-MS-Has-Attach: 16, 23 -- X-MS-TNEF-Correlator: 16, 23 -- Thread-Topic: [Microscopy] dbl stick carbon tab blues 16, 23 -- Thread-Index: AcZ46A6qIDfGYibgSqi1YHWKtSO+WwAA5z8g 16, 23 -- From: "James Chalcroft" {jchalcro-at-neuro.mpg.de} 16, 23 -- To: {opmills-at-mtu.edu} 16, 23 -- Cc: {Microscopy-at-microscopy.com} 16, 23 -- Content-Transfer-Encoding: 8bit 16, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4GDQEre001944 ==============================End of - Headers==============================
-----Original Message----- X-from: opmills-at-mtu.edu [mailto:opmills-at-mtu.edu] Sent: Tuesday, May 16, 2006 2:50 PM To: j.bilde-at-risoe.dk
Several years ago many (most?) of the EM supply vendors switched the formula for carbon tabs to a "stiff", non-flexible formula. I was told at that time that the old formula used typewriter tape as a component.
These new stiff tabs are unsuitable for many of the preps we do. I was able to find the old formula at Fullam and now they have switched too.
Does anyone know where to find truly flexible carbon tabs. The ones Fullam had were from Nisshin EM, very expensive but quite high quality.
Thanks in advance.
Owen Mills MTU, Houghton MI
==============================Original Headers============================== 5, 31 -- From opmills-at-mtu.edu Tue May 16 07:48:57 2006 5, 31 -- Received: from node7.mtu.edu (node7.mtu.edu [141.219.69.7]) 5, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GCmvf9024068 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 07:48:57 -0500 5, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 5, 31 -- by node7.mtu.edu (8.13.1/8.13.1) with ESMTP id k4GCmu39024410 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:56 -0400 5, 31 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 5, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k4GCmuco022544 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:56 -0400 5, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) 5, 31 -- by node34.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k4GCmu6b023709 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:56 -0400 5, 31 -- (envelope-from opmills-at-mtu.edu) 5, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 5, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k4GCmtjx010574 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:55 -0400 (EDT) 5, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 5, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k4GCmtRx016397 5, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:48:55 -0400 5, 31 -- Mime-Version: 1.0 (Apple Message framework v749.3) 5, 31 -- Content-Transfer-Encoding: 7bit 5, 31 -- Message-Id: {E9327AEB-33E3-4E6C-9DA6-F2AA87734FF8-at-mtu.edu} 5, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 5, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 5, 31 -- Subject: dbl stick carbon tab blues 5, 31 -- Date: Tue, 16 May 2006 08:49:15 -0400 5, 31 -- X-Mailer: Apple Mail (2.749.3) 5, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.5.16.53106 5, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__CP_MEDIA_2_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
==============================Original Headers============================== 18, 24 -- From j.bilde-at-risoe.dk Tue May 16 08:29:28 2006 18, 24 -- Received: from mail.risoe.dk (mail.risoe.dk [130.226.48.21]) 18, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GDTS2l007866 18, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:29:28 -0500 18, 24 -- Received: from EXCHG-VS1.risoe.dk (exchg-01.risoe.dk [10.0.0.26]) 18, 24 -- by risoe.dk (PMDF V6.2-X26 #31094) with ESMTP id {ZHVS08QY0HQOW604L8-at-risoe.dk} 18, 24 -- for Microscopy-at-microscopy.com; Tue, 18, 24 -- 16 May 2006 15:29:05 +0200 (Romance Daylight Time) 18, 24 -- Date: Tue, 16 May 2006 15:28:16 +0200 18, 24 -- From: j.bilde-at-risoe.dk 18, 24 -- Subject: RE: [Microscopy] dbl stick carbon tab blues 18, 24 -- To: opmills-at-mtu.edu 18, 24 -- Cc: Microscopy-at-microscopy.com 18, 24 -- Message-id: {6463F256A85DC14CAB07F087A247997232D4E0-at-EXCHG-VS1.risoe.dk} 18, 24 -- MIME-version: 1.0 18, 24 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 18, 24 -- Content-type: text/plain; charset=iso-8859-1 18, 24 -- Thread-Topic: [Microscopy] dbl stick carbon tab blues 18, 24 -- Thread-Index: AcZ454gbgA//L9RHSESEcXhV+9r4twAA62Ug 18, 24 -- Content-class: urn:content-classes:message 18, 24 -- X-MS-Has-Attach: 18, 24 -- X-MS-TNEF-Correlator: 18, 24 -- Content-Transfer-Encoding: 8bit 18, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4GDTS2l007866 ==============================End of - Headers==============================
I suppose that you have not only a cold FEG SEM, but a so called "semi-in-lens" type of OL on it too. In fact the cold or schottky type of FEG doesn't play a role in the problem. It's only a OL question. That type of OL has a strong magnetic field coming out of it, which enveloppe the sample at short working distance, alouding to work virtuusely at WD0 and to get nice high res images of ..... non magnetic materials, with the "trough the lens" SE detector. The field coming out of the OL can be very strong ; I have measured values such as 3kG at WD2 and 10keV primary energie. With lower beam energy the field decreases and at 3keV, I measured a value of 1.2kG at WD3 mm, and 0.3kG at WD8 mm.
Pratically, you have a few solutions, but all are half solutions.
First, you must avoid to work with short WD. Look at the shortes WD possible without too much astigmatisme. Ask the SEM manufacturer at which WD the sample is quite out of the field. If your shortest WD is 2mm or so, it will bee something like 6-8mm.
Second, you must find a primary energie not to low, which would give a poor resolution, due to the perturbation of the primary beam by the field lignes in the sample, and not too high, to minimase the filed coming out of the OL, wich increases with increasing primary energy. In most cases, you 'll find a good compromise between 5 and 10 keV.
Third, you must play very very much with the astigmatism corrections, at replay again if you move your sample a little bit. And depending of the electronic, you may touch the range limits of the astigmatism corrections. In that case, if you have a CL astigmatism correction setting, you can play on it a little bit, puting astigmatisme at the Cl, in the opposit direction, to gain some margin of the OL astigmatism settings. It's not a clean way to work, but in may help.
Fourth, take the smalest sample possible, not too thick, not too wide, and fix it very securly on the holder. It may land from it and stick to the OL !
Fifth solutions : buy an other SEM for that kind of samples !!!
Don't wait as well resolved images than with ..... gold particles on silicon. If you have a nice picture at x50000, you can be satisfied ! More depends on your particular situation, sample and SEM.
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
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==============================Original Headers============================== 17, 30 -- From jacques.faerber-at-ipcms.u-strasbg.fr Tue May 16 08:50:33 2006 17, 30 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.157]) 17, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GDoX8l022053 17, 30 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 08:50:33 -0500 17, 30 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 17, 30 -- by mailhost.u-strasbg.fr (8.13.4/jtpda-5.5pre1) with ESMTP id k4GDoRKQ074518 17, 30 -- ; Tue, 16 May 2006 15:50:27 +0200 (CEST) 17, 30 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr [130.79.54.3]) 17, 30 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id CA41810000E6; 17, 30 -- Tue, 16 May 2006 15:50:15 +0200 (CEST) 17, 30 -- Message-ID: {4469D88A.7030305-at-ipcms.u-strasbg.fr} 17, 30 -- Date: Tue, 16 May 2006 15:50:02 +0200 17, 30 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 17, 30 -- User-Agent: Mozilla Thunderbird 1.0.8 (X11/20060503) 17, 30 -- X-Accept-Language: fr, en 17, 30 -- MIME-Version: 1.0 17, 30 -- To: lewryan-at-gmail.com, Microscopy-at-microscopy.com 17, 30 -- Subject: Re: [Microscopy] viaWWW: SEM imaging on sample with magnetic substrate 17, 30 -- References: {200605152208.k4FM88SO002253-at-ns.microscopy.com} 17, 30 -- In-Reply-To: {200605152208.k4FM88SO002253-at-ns.microscopy.com} 17, 30 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 17, 30 -- Content-Transfer-Encoding: 8bit 17, 30 -- X-IPCMS-MailScanner: Found to be clean 17, 30 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 17, 30 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.157]); Tue, 16 May 2006 15:50:27 +0200 (CEST) 17, 30 -- X-Virus-Scanned: ClamAV 0.88.1/1464/Tue May 16 11:36:19 2006 on mr7.u-strasbg.fr 17, 30 -- X-Virus-Status: Clean 17, 30 -- X-Spam-Status: No, score=-1.4 required=5.0 tests=ALL_TRUSTED,AWL 17, 30 -- autolearn=disabled version=3.1.1 17, 30 -- X-Spam-Checker-Version: SpamAssassin 3.1.1 (2006-03-10) on mr7.u-strasbg.fr ==============================End of - Headers==============================
I have a faculty member at UIC who is looking for a one off image of a purified protein. She has seen this done by rotary shadowing and while we have the instrumentation at the university we do not have the necessary experience. As it is a one off experiment (she needs an image showing to confirm if there is bend in the middle of a mutant) I was wondering if there was a lab out there that regularly uses rotary shadowing who could help us?
Alan
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
We have used the AFM to image DNA conformation and think that it might also work for your protein, without all the elaborate preparation.
If you are interested in having us give it a try, contact Dr. Kim Kangasniemi (just call him "Kim") at (972)954-8014 or kim-at-nt-america.com
Hope this was helpful,
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through 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 08:54 AM 5/16/2006, nicholls-at-uic.edu wrote:
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==============================Original Headers============================== 15, 17 -- From bfoster-at-mme1.com Tue May 16 10:18:21 2006 15, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 15, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GFIKxw011149 15, 17 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 10:18:20 -0500 15, 17 -- Received: (qmail 9713 invoked by uid 2020); 16 May 2006 10:34:25 -0500 15, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 15, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 16 May 2006 10:34:24 -0500 15, 17 -- Message-Id: {7.0.1.0.0.20060516101501.020b2278-at-mme1.com} 15, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 17 -- Date: Tue, 16 May 2006 10:18:26 -0500 15, 17 -- To: nicholls-at-uic.edu, microscopy Listserver {microscopy-at-microscopy.com} 15, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 15, 17 -- Subject: Re: [Microscopy] Rotary Shadowing 15, 17 -- In-Reply-To: {200605161354.k4GDssi4001193-at-ns.microscopy.com} 15, 17 -- References: {200605161354.k4GDssi4001193-at-ns.microscopy.com} 15, 17 -- Mime-Version: 1.0 15, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Ted Pella has good ones as does EMS. No experience with SPI tabs.
gary g.
At 05:51 AM 5/16/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Tue May 16 11:27:57 2006 10, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4GGRu3l022232 10, 20 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 11:27:57 -0500 10, 20 -- Received: (qmail 704 invoked from network); 16 May 2006 09:27:56 -0700 10, 20 -- Received: by simscan 1.1.0 ppid: 701, pid: 702, t: 0.1669s 10, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 20 -- by qsmtp3 with SMTP; 16 May 2006 09:27:56 -0700 10, 20 -- Message-Id: {7.0.1.0.2.20060516092606.023c7328-at-gaugler.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 20 -- Date: Tue, 16 May 2006 09:27:33 -0700 10, 20 -- To: opmills-at-mtu.edu 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] dbl stick carbon tab blues 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200605161251.k4GCpEVf027234-at-ns.microscopy.com} 10, 20 -- References: {200605161251.k4GCpEVf027234-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
We have been doing our pre-embedding immunogold localizations incubations for animal tissue in PBS, 0.2% BSA and 10mM Na Azide.
I am working now on some em pre-embedding localizations on plant tissue, for which light microscopy localizations have been done using MTSB buffer (50mM PIPES, 5mM MgSO4, 5mM EGTA, pH~7.0). Is there any reason for which I should not be using this same MTSB buffer for the em work?
Thank you very much for your suggestions. They will definitely save me trial time.
Thanks.
Tea
-- *************************************** Tea Meulia, PhD Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
tel.: 330-263-3836 or -3828 fax: 330-202-3563
http://www.oardc.ohio-state.edu/mcic
*****************************************
==============================Original Headers============================== 11, 18 -- From meulia.1-at-osu.edu Tue May 16 13:17:06 2006 11, 18 -- Received: from defang9.net.ohio-state.edu (defang9.net.ohio-state.edu [128.146.216.78]) 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GIH5xS001081 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 13:17:05 -0500 11, 18 -- Received: from [164.107.85.164] (dhcp-107-85-164.oardc.ohio-state.edu [164.107.85.164]) 11, 18 -- by defang9.net.ohio-state.edu (8.13.1/8.13.1) with ESMTP id k4GIH5AC031731 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 14:17:05 -0400 11, 18 -- Mime-Version: 1.0 11, 18 -- X-Sender: meulia.1-at-pop.service.ohio-state.edu 11, 18 -- Message-Id: {p05200f03c08fc56f761a-at-[164.107.85.164]} 11, 18 -- Date: Tue, 16 May 2006 14:17:12 -0400 11, 18 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 11, 18 -- From: Tea Meulia {meulia.1-at-osu.edu} 11, 18 -- Subject: em immunolocalizations 11, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 11, 18 -- X-Spam-Score: undef - spam scanning disabled 11, 18 -- X-CanItPRO-Stream: outbound 11, 18 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 128.146.216.12 ==============================End of - Headers==============================
Hi, In dealing with plant tissue, EGTA is included in buffers because calcium chelation removes calcium cross bridges and probably some pectin from the cell wall and allows antiboody access. Although protocols for doing this originally called for having EGTA in the fixation buffer, in our hands, we get better preservation if the EGTA is included after the fixation as a separate incubation. The removal of the calcium by the same token makes the cell wall weaker. You may find quite distorted tissue. It may be possible to minimize this distortion by including an incubation in mM CaCl2 after the 2nd antibody and before dehydration.
Note that buffers with pipes, magnesium, and EGTA are not microtubule stablizing (if that is what you mean by MTSB). They are the standard buffers for studying microtubule dynamics in vitro.
Hope this helps.
Tobias } } } We have been doing our pre-embedding immunogold localizations } incubations for animal tissue in PBS, 0.2% BSA and 10mM Na Azide. } } I am working now on some em pre-embedding localizations on plant } tissue, for which light microscopy localizations have been done using } MTSB buffer (50mM PIPES, 5mM MgSO4, 5mM EGTA, pH~7.0). Is there any } reason for which I should not be using this same MTSB buffer for the } em work? } } Thank you very much for your suggestions. They will definitely save } me trial time. } } Thanks. } } Tea } } } } -- } *************************************** } Tea Meulia, PhD } Director } Molecular and Cellular Imaging Center } Ohio State University/OARDC } 1680 Madison Ave. } Wooster OH 44691 } } tel.: 330-263-3836 or -3828 } fax: 330-202-3563 } } http://www.oardc.ohio-state.edu/mcic } } ***************************************** } } ==============================Original Headers============================== } 11, 18 -- From meulia.1-at-osu.edu Tue May 16 13:17:06 2006 } 11, 18 -- Received: from defang9.net.ohio-state.edu } (defang9.net.ohio-state.edu [128.146.216.78]) } 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k4GIH5xS001081 } 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 } 13:17:05 -0500 } 11, 18 -- Received: from [164.107.85.164] } (dhcp-107-85-164.oardc.ohio-state.edu [164.107.85.164]) } 11, 18 -- by defang9.net.ohio-state.edu (8.13.1/8.13.1) with } ESMTP id k4GIH5AC031731 } 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 } 14:17:05 -0400 } 11, 18 -- Mime-Version: 1.0 } 11, 18 -- X-Sender: meulia.1-at-pop.service.ohio-state.edu } 11, 18 -- Message-Id: {p05200f03c08fc56f761a-at-[164.107.85.164]} } 11, 18 -- Date: Tue, 16 May 2006 14:17:12 -0400 } 11, 18 -- To: Microscopy Listserver {microscopy-at-microscopy.com} } 11, 18 -- From: Tea Meulia {meulia.1-at-osu.edu} } 11, 18 -- Subject: em immunolocalizations } 11, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 11, 18 -- X-Spam-Score: undef - spam scanning disabled } 11, 18 -- X-CanItPRO-Stream: outbound } 11, 18 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 128.146.216.12 } ==============================End of - Headers==============================
I would suggest setting the specimen at a working distance of at least 15mm so as to be outside the field of the lens. What may also help is to lower the kV as this will also lower the lens field. Of course the kV level will depend upon what you are asking of the specimen?
If you wish to examine the TRUE surface {5kV will be ideal. If you wish to investigate the sub surface detail 15kV backscatter would probably be a good starting point.
Do not try to use an upper detector if fitted tot he instrument as this will require you being close to the lens. Lower detectors in a twin detector system require a higher probe current (weaker C1) than that used for an upper detector.
Good luck
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {lewryan-at-gmail.com} To: {protrain-at-emcourses.com} Sent: Monday, May 15, 2006 11:04 PM
I need to look at some RBC's using SEM. anyone have a favorite protocol for preparing them? Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
We've used poly-l-lysine cover slips and standard fixation/dehydration procedures with good success. I believe some people have used HMDS successfully, too.
Randy
-----Original Message----- X-from: phillipst-at-missouri.edu [mailto:phillipst-at-missouri.edu] Sent: Tuesday, May 16, 2006 1:55 PM To: Tindall, Randy D.
I need to look at some RBC's using SEM. anyone have a favorite protocol for preparing them? Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Someone wants to send me isolated, fixed rhinovirus on a grid for negative staining and imaging. I have not worked with rhinovirus before. Does anyone have a favorite protocol for fixation and staining? Glut or PFA? Uranyl acetate or PTA?
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============================== 5, 19 -- From tina-at-pbrc.hawaii.edu Tue May 16 14:32:01 2006 5, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GJVxtS019267 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 May 2006 14:32:00 -0500 5, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 5, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k4GJVs2a020635 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 May 2006 09:31:55 -1000 (HST) 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k4GJVrsk020630 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 May 2006 09:31:53 -1000 (HST) 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 5, 19 -- Date: Tue, 16 May 2006 09:31:52 -1000 (HST) 5, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 5, 19 -- X-Sender: tina-at-halia 5, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 5, 19 -- Subject: Rhinovirus negative stain 5, 19 -- Message-ID: {Pine.GSO.4.21.0605160929390.20461-100000-at-halia} 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Sorry to bring this topic up again. I know it has been discussed:
What are the additives that can go in a water recirculator for a TEM? I use to use a product called cool-prep and someone said ethylene glycol in the same ratio. Any thoughts on that?
thanks, 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)
I've got a user who wants to characterize 60-atom Carbon Buckyballs. Now He wants to start by determining if they clump, form larger groupsing etc. when mixed into ultra pure water (Q- water), and then other quality waters. So, any one out there have any idea on how to determine simple sizing? In a hydrated state? In suspension? At potentially nanoscale? Obviously, going EM requires drying them down - which artifically modifys the samples. Cryo- might work but we don't have the capability here. SPM might work but how to adhere to a surface for imaging? Diffraction of some sort?
Any ideas?
Thanks in advance!
Richard E. Edelmann, Ph.D. Associate Editor of EXPO, Microscopy and Microanalysis Supplement Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
==============================Original Headers============================== 5, 23 -- From edelmare-at-muohio.edu Tue May 16 15:32:16 2006 5, 23 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GKWG5v013777 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 16 May 2006 15:32:16 -0500 5, 23 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 5, 23 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k4GKWEox022092 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 16 May 2006 16:32:14 -0400 5, 23 -- Received: from emf03 ([134.53.14.97]) 5, 23 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.6) with ESMTP id k4GKWDBu018999 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 16 May 2006 16:32:13 -0400 5, 23 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 5, 23 -- To: microscopy-at-Microscopy.com 5, 23 -- Date: Tue, 16 May 2006 16:39:28 -0400 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-type: text/plain; charset=US-ASCII 5, 23 -- Content-transfer-encoding: 7BIT 5, 23 -- Subject: Sizing 60-atom bucky balls 5, 23 -- Message-ID: {446A0040.18005.1D72926-at-localhost} 5, 23 -- Priority: normal 5, 23 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 5, 23 -- X-Real-ConnectIP: 134.53.14.97 5, 23 -- X-Scanned-By: MIMEDefang 2.45 5, 23 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 ==============================End of - Headers==============================
Nestor, Finally a question I can answer for someone! RBC = Red Blood Cell (~7 microns or so in size)
Not to be confused with TLA (three letter acronym) or DAP (Parents against Dyslexia) ;o)
Paul
---------------------------------------------------------------- Though the boys throw stones at the frogs in sport, the frogs do not die in sport, but in earnest. - Plutarch
==============================Original Headers============================== 5, 21 -- From pgrover-at-bilbo.bio.purdue.edu Tue May 16 15:33:15 2006 5, 21 -- Received: from mailhub129.itcs.purdue.edu (mailhub129.itcs.purdue.edu [128.210.5.129]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GKXFi0016952 5, 21 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 15:33:15 -0500 5, 21 -- Received: from paklabpgrover (dhcp155-122.bio.purdue.edu [128.210.155.122]) 5, 21 -- by mailhub129.itcs.purdue.edu (8.13.6/8.13.4/internal-smtp) with ESMTP id k4GKXErl018860 5, 21 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 16:33:15 -0400 5, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} 5, 21 -- To: {microscopy-at-microscopy.com} 5, 21 -- Subject: re: RBC 5, 21 -- Date: Tue, 16 May 2006 16:33:18 -0400 5, 21 -- Message-ID: {000001c67927$f7957350$7a9bd280-at-paklabpgrover} 5, 21 -- MIME-Version: 1.0 5, 21 -- Content-Type: text/plain; 5, 21 -- charset="us-ascii" 5, 21 -- Content-Transfer-Encoding: 7bit 5, 21 -- X-Mailer: Microsoft Office Outlook 11 5, 21 -- Thread-Index: AcZ5J/dsq3a+geObQAWpSodYpvluUQ== 5, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 5, 21 -- X-PMX-Version: 5.1.2.240295 5, 21 -- X-PerlMx-Virus-Scanned: Yes ==============================End of - Headers==============================
I use deionized water only (no additives) and have had no problems over the past twenty years. An important note is that we prevent the growth of algae by plumbing all our chillers and microscopes with opaque water hose. We never have algae, or any other, growth in our chillers.
Regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
beth-at-plantbio. uga.edu To gary.m.brown-at-exxonmobil.com 05/16/06 03:24 cc PM Subject [Microscopy] TEM water recirculator Please respond additives to beth-at-plantbio. uga.edu
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Sorry to bring this topic up again. I know it has been discussed:
What are the additives that can go in a water recirculator for a TEM? I use to use a product called cool-prep and someone said ethylene glycol in the same ratio. Any thoughts on that?
thanks, 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)
I would suggest small-angle x-ray scattering (SAXS). Can be done hydrated.
See, for example:
http://www.uni.aps.anl.gov/usaxs/
Jeffrey A. Fortner, Ph.D. Chemical Engineering Division Argonne National Laboratory Argonne, IL 60439 fortner(at)cmt.anl.gov
-----Original Message----- X-from: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] Sent: Tuesday, May 16, 2006 3:33 PM To: Fortner, Jeffrey A.
O.k., folks here's an odd problem:
I've got a user who wants to characterize 60-atom Carbon Buckyballs. Now He wants to start by determining if they clump, form larger groupsing etc. when mixed into ultra pure water (Q- water), and then other quality waters. So, any one out there have any idea on how to determine simple sizing? In a hydrated state? In suspension? At potentially nanoscale? Obviously, going EM requires drying them down - which artifically modifys the samples. Cryo- might work but we don't have the capability here. SPM might work but how to adhere to a surface for imaging? Diffraction of some sort?
Any ideas?
Thanks in advance!
Richard E. Edelmann, Ph.D. Associate Editor of EXPO, Microscopy and Microanalysis Supplement Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
==============================Original Headers============================== 5, 23 -- From edelmare-at-muohio.edu Tue May 16 15:32:16 2006 5, 23 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GKWG5v013777 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 16 May 2006 15:32:16 -0500 5, 23 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 5, 23 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k4GKWEox022092 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 16 May 2006 16:32:14 -0400 5, 23 -- Received: from emf03 ([134.53.14.97]) 5, 23 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.6) with ESMTP id k4GKWDBu018999 5, 23 -- for {microscopy-at-Microscopy.com} ; Tue, 16 May 2006 16:32:13 -0400 5, 23 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 5, 23 -- To: microscopy-at-Microscopy.com 5, 23 -- Date: Tue, 16 May 2006 16:39:28 -0400 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-type: text/plain; charset=US-ASCII 5, 23 -- Content-transfer-encoding: 7BIT 5, 23 --
Nester,
RBC is the acronym for "red blood cells".
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
zaluzec-at-aaem.a mc.anl.gov To gary.m.brown-at-exxonmobil.com 05/16/06 03:31 cc PM Subject [Microscopy] Hmmm... what is RBC Please respond to zaluzec-at-aaem.a mc.anl.gov
Human red blood cells respond nicely to fairly routine fixatives. We use cacodylate with Ca, Mg, and NaCl added as buffer system but biological buffers such as PIPES should work also.
However, be aware the RBC's from other animals have different osmotic requirements and may crenellate easily when the H-RBCs are fine. We ran into real problems with mouse RBC's a while back. Only sure way to avoid problems is to check and balance osmolarity of fixatives for the specific host RBCs.
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: {TindallR-at-missouri.edu} } Reply-To: {TindallR-at-missouri.edu} } Date: Tue, 16 May 2006 14:11:02 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] RE: SEM of RBC's } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Tom, } } We've used poly-l-lysine cover slips and standard fixation/dehydration } procedures with good success. I believe some people have used HMDS } successfully, too. } } Randy } } -----Original Message----- } X-from: phillipst-at-missouri.edu [mailto:phillipst-at-missouri.edu] } Sent: Tuesday, May 16, 2006 1:55 PM } To: Tindall, Randy D. } Subject: [Microscopy] SEM of RBC's } } } } } ------------------------------------------------------------------------ } ---- } 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 need to look at some RBC's using SEM. anyone have a favorite protocol } for preparing them? Thanks, Tom } } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 2 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } } } ==============================Original } Headers============================== } 7, 18 -- From PhillipsT-at-missouri.edu Tue May 16 13:54:11 2006 } 7, 18 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k4GIsAWG019726 } 7, 18 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 16 May 2006 } 13:54:10 -0500 } 7, 18 -- Received: from um-exproto9.um.umsystem.edu ([207.160.151.148]) } by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 7, 18 -- Tue, 16 May 2006 13:54:07 -0500 } 7, 18 -- Received: from phillips-dell.missouri.edu ([128.206.81.132]) by } um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft } SMTPSVC(6.0.3790.1830); } 7, 18 -- Tue, 16 May 2006 13:54:06 -0500 } 7, 18 -- Message-Id: } {6.0.0.22.2.20060516135251.01cc6e50-at-pop.missouri.edu} } 7, 18 -- X-Sender: phillipst-at-pop.missouri.edu } 7, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 6.0.0.22 } 7, 18 -- Date: Tue, 16 May 2006 13:54:06 -0500 } 7, 18 -- To: Microscopy-at-msa.microscopy.com } 7, 18 -- From: Tom Phillips {phillipst-at-missouri.edu} } 7, 18 -- Subject: SEM of RBC's } 7, 18 -- Mime-Version: 1.0 } 7, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } 7, 18 -- X-OriginalArrivalTime: 16 May 2006 18:54:06.0810 (UTC) } FILETIME=[1BBCB3A0:01C6791A] } ==============================End of - } Headers============================== } } } ==============================Original Headers============================== } 16, 24 -- From TindallR-at-missouri.edu Tue May 16 14:08:17 2006 } 16, 24 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4GJ8GxK008815 } 16, 24 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 14:08:17 -0500 } 16, 24 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by } um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 16, 24 -- Tue, 16 May 2006 14:08:16 -0500 } 16, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 16, 24 -- Content-class: urn:content-classes:message } 16, 24 -- MIME-Version: 1.0 } 16, 24 -- Content-Type: text/plain; } 16, 24 -- charset="US-ASCII" } 16, 24 -- Subject: RE: [Microscopy] SEM of RBC's } 16, 24 -- Date: Tue, 16 May 2006 14:08:16 -0500 } 16, 24 -- Message-ID: } {BA876152E8653240BE8572E897083EE701849FC4-at-UM-EMAIL09.um.umsystem.edu} } 16, 24 -- X-MS-Has-Attach: } 16, 24 -- X-MS-TNEF-Correlator: } 16, 24 -- Thread-Topic: [Microscopy] SEM of RBC's } 16, 24 -- thread-index: AcZ5Gj/IXaM4jxnBT36YosCHI/gn+wAAa8yQ } 16, 24 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 16, 24 -- To: "Phillips, Thomas E." {PhillipsT-at-missouri.edu} } 16, 24 -- Cc: {microscopy-at-microscopy.com} } 16, 24 -- X-OriginalArrivalTime: 16 May 2006 19:08:16.0800 (UTC) } FILETIME=[165EE200:01C6791C] } 16, 24 -- Content-Transfer-Encoding: 8bit } 16, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k4GJ8GxK008815 } ==============================End of - Headers==============================
==============================Original Headers============================== 7, 23 -- From dsherman-at-purdue.edu Tue May 16 15:54:16 2006 7, 23 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GKsGb8002076 7, 23 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 15:54:16 -0500 7, 23 -- Received: from exchange.purdue.edu ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 23 -- Tue, 16 May 2006 16:54:15 -0400 7, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 7, 23 -- Tue, 16 May 2006 20:54:15 +0000 7, 23 -- User-Agent: Microsoft-Entourage/11.2.3.060209 7, 23 -- Date: Tue, 16 May 2006 16:54:14 -0400 7, 23 -- Subject: Re: [Microscopy] RE: SEM of RBC's 7, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 23 -- To: Randy Tindall {TindallR-at-missouri.edu} , 7, 23 -- "message to: MSA list" {microscopy-at-microscopy.com} 7, 23 -- Message-ID: {C08FB436.10893%dsherman-at-purdue.edu} 7, 23 -- Thread-Topic: [Microscopy] RE: SEM of RBC's 7, 23 -- Thread-Index: AcZ5KuOOIlbu4uUeEdq+igARJN08Mg== 7, 23 -- In-Reply-To: {200605161911.k4GJB2I3012913-at-ns.microscopy.com} 7, 23 -- Mime-version: 1.0 7, 23 -- Content-type: text/plain; 7, 23 -- charset="US-ASCII" 7, 23 -- Content-transfer-encoding: 7bit 7, 23 -- X-OriginalArrivalTime: 16 May 2006 20:54:15.0879 (UTC) FILETIME=[E4AD5570:01C6792A] ==============================End of - Headers==============================
On May 16, 2006, at 1:22 PM, beth-at-plantbio.uga.edu wrote:
} Sorry to bring this topic up again. I know it has been discussed: } } What are the additives that can go in a water recirculator for a TEM? } I use to use a product called cool-prep and someone said ethylene } glycol in the same ratio. } Any thoughts on that? } Dear Beth, I don't know what is in cool-prep, but if ethylene glycol is a good substitute, it sounds like anti-freeze. The important things for a TEM water additive are to prevent corrosion and growth of bacteria and algae, but the cooling water is not so cold that anti-freeze is necessary. I have added a molybdenum-based corrosion inhibitor and the chemical 2,2'-Methylenebis(4-chloro-phenol), AKA dichlorophene, to inhibit growth with good effect. It is also important to keep the pH at ~8 to prevent Cu from dissolving. 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 May 16 15:59:24 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 k4GKxOnP011992 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 16 May 2006 15:59:24 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id C0BF736587 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 16 May 2006 13:59:23 -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 636B510B473 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 16 May 2006 13:59:01 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200605162022.k4GKMSWF030299-at-ns.microscopy.com} 4, 22 -- References: {200605162022.k4GKMSWF030299-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {9b9f76870a81061eedc92a3f407cecae-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] TEM water recirculator additives 4, 22 -- Date: Tue, 16 May 2006 14:09:49 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Thanks, all, for the suggestions about negative staining of rhinovirus.
With one notorious exception, I have never had to fix virus before staining, but these people want to send me fixed rhinovirus on commercial non-glow-discharged grids (sigh). And it looks like I'll try my usual stable of stains; UrAc, PTA, and NH4M.
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 Tue May 16 17:28:49 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 k4GMSmTE000510 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 May 2006 17:28:49 -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 k4GMSjGY022033 6, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 May 2006 12:28:46 -1000 (HST) 6, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k4GMSjrc022030 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 May 2006 12:28:45 -1000 (HST) 6, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 6, 19 -- Date: Tue, 16 May 2006 12:28:44 -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: Rhinovirus - thanks 6, 19 -- Message-ID: {Pine.GSO.4.21.0605161225040.21962-100000-at-halia} 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
On May 16, 2006, at 2:50 PM, Yang, Ann-Fook wrote:
} Please let us know how much you put in. Thank you. } Hi Ann-Fook, I aim for 100 ppm of the Mo, as determined by a test kit, K1805, from Taylor Technonogies, Inc. This is somewhat complicated due to not knowing how much water is in the Haskris and how much is in the tubing, lenses, etc., so I add a few hundred ml of the corrosion inhibitor, check it monthly, and add more when necessary. After a few months, you will get a pretty good idea of how much inhibitor it takes to increase the ppm Mo by a given amount. I just float a small amount of the dichlorophene on top of the water in the Haskris--it's not very soluble--and add more when little solid remains. Yours, Bill
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue May 16 17:32:34 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 k4GMWY3i006767 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 16 May 2006 17:32:34 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP 4, 22 -- id C3ABC34F3D; Tue, 16 May 2006 15:32:33 -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 D0245354CB; Tue, 16 May 2006 15:24:46 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {E035A9C87303AE4AB9BA10FD8324DFB1024334EC-at-onncrxms3.agr.gc.ca} 4, 22 -- References: {E035A9C87303AE4AB9BA10FD8324DFB1024334EC-at-onncrxms3.agr.gc.ca} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {c8fa41ce51653c0128aaf93b4f011bed-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Re: TEM water recirculator additives 4, 22 -- Date: Tue, 16 May 2006 15:35:35 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com, "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Just wanted to thank everyone for the responses to my posting about the TEM water chiller additives. I'll try just using distilled water and monitoring the system more frequently. Thank you! 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)
Quite a while ago now I did some pre-embedding immunogold labelling, and fixed in phosphate rather than Pipes buffer, using 2 mM rather than 5 mM Mg and EGTA. I think use the fixative that works best for what you're after, I'd suggest using the same fixative/buffer combination as for the LM work, taking on board Tobias' comments about how EGTA softens the walls causing some tissue distortion if you're not careful.
When I did this, I then cut frozen sections, rinsed in PBS then labelled the sections on slides with antibodies in PBS, after the usual blocking in BSA or gelatin. Then embedded the sections in Spurr's (messy) before sectioning for TEM. I did this to get greater penetration of label into tissue, while avoiding the original cut surface of the tissue block.
good luck, cheers, Rosemary
Rosemary White rosemary.white-at-csiro.au CSIRO Plant Industry ph. 02-6246 5475 GPO Box 1600 mob. 0402 835 973 Canberra, ACT 2601 fax. 02-6246 5334 Australia
} From: meulia.1-at-osu.edu } Reply-To: meulia.1-at-osu.edu } Date: Tue, 16 May 2006 13:19:55 -0500 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] em immunolocalizations } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } We have been doing our pre-embedding immunogold localizations } incubations for animal tissue in PBS, 0.2% BSA and 10mM Na Azide. } } I am working now on some em pre-embedding localizations on plant } tissue, for which light microscopy localizations have been done using } MTSB buffer (50mM PIPES, 5mM MgSO4, 5mM EGTA, pH~7.0). Is there any } reason for which I should not be using this same MTSB buffer for the } em work? } } Thank you very much for your suggestions. They will definitely save } me trial time. } } Thanks. } } Tea } } } } -- } *************************************** } Tea Meulia, PhD } Director } Molecular and Cellular Imaging Center } Ohio State University/OARDC } 1680 Madison Ave. } Wooster OH 44691 } } tel.: 330-263-3836 or -3828 } fax: 330-202-3563 } } http://www.oardc.ohio-state.edu/mcic } } ***************************************** } } ==============================Original Headers============================== } 11, 18 -- From meulia.1-at-osu.edu Tue May 16 13:17:06 2006 } 11, 18 -- Received: from defang9.net.ohio-state.edu } (defang9.net.ohio-state.edu [128.146.216.78]) } 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4GIH5xS001081 } 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 13:17:05 -0500 } 11, 18 -- Received: from [164.107.85.164] } (dhcp-107-85-164.oardc.ohio-state.edu [164.107.85.164]) } 11, 18 -- by defang9.net.ohio-state.edu (8.13.1/8.13.1) with ESMTP id } k4GIH5AC031731 } 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 14:17:05 -0400 } 11, 18 -- Mime-Version: 1.0 } 11, 18 -- X-Sender: meulia.1-at-pop.service.ohio-state.edu } 11, 18 -- Message-Id: {p05200f03c08fc56f761a-at-[164.107.85.164]} } 11, 18 -- Date: Tue, 16 May 2006 14:17:12 -0400 } 11, 18 -- To: Microscopy Listserver {microscopy-at-microscopy.com} } 11, 18 -- From: Tea Meulia {meulia.1-at-osu.edu} } 11, 18 -- Subject: em immunolocalizations } 11, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 11, 18 -- X-Spam-Score: undef - spam scanning disabled } 11, 18 -- X-CanItPRO-Stream: outbound } 11, 18 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 128.146.216.12 } ==============================End of - Headers============================== }
==============================Original Headers============================== 7, 22 -- From Rosemary.White-at-csiro.au Tue May 16 17:51:17 2006 7, 22 -- Received: from act-MTAout6.csiro.au (act-MTAout6.csiro.au [150.229.7.43]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4GMpGIO030218 7, 22 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 16 May 2006 17:51:17 -0500 7, 22 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=CtH5eq0n7NhwH/zu+AOFn0CNv9fLVrTMaTXnfBEZo4kYpLhxiFeD2RgI+/wLp0iJUo8yaVMHLXIZLatlQlztjp/7BsIH8dZDFC/GmX6TQ2f9X94vVHhYppLtYxShzvdG; 7, 22 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 7, 22 -- by act-MTAout6.csiro.au with ESMTP; 17 May 2006 08:51:15 +1000 7, 22 -- X-IronPort-AV: i="4.05,135,1146405600"; 7, 22 -- d="scan'208"; a="94577057:sNHT25900424" 7, 22 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 7, 22 -- Wed, 17 May 2006 08:51:14 +1000 7, 22 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 7, 22 -- Date: Wed, 17 May 2006 08:53:23 +1000 7, 22 -- Subject: Re: [Microscopy] em immunolocalizations 7, 22 -- From: Rosemary White {Rosemary.White-at-csiro.au} 7, 22 -- To: {Microscopy-at-msa.microscopy.com} 7, 22 -- Message-ID: {C0909503.167AA%Rosemary.White-at-csiro.au} 7, 22 -- In-Reply-To: {200605161819.k4GIJtIV005765-at-ns.microscopy.com} 7, 22 -- Mime-version: 1.0 7, 22 -- Content-type: text/plain; charset="US-ASCII" 7, 22 -- Content-transfer-encoding: 7bit 7, 22 -- X-OriginalArrivalTime: 16 May 2006 22:51:14.0869 (UTC) FILETIME=[3C523650:01C6793B] ==============================End of - Headers==============================
The buffer you indicated may be fine, although a word of warning may be in place: be careful when using divalent cations like Mg2+ . We've never actually tested this for gold conjugates but such ions cause aggregates of gold particles even at very low concentrations. The coating proteins should help preventing that but it may still happen. If that buffer, as Tobias Baskin indicates, is used to open cell walls to antibodies, then it may be sufficient if it is applied only to obtain that effect, i.e. fix, treat with the permeabilising buffer and then wash with PBS a few times before proceeding using the same protocol that was used for your animal tissue.
Good luck
Jan
On May 17, 2006, at 6:17 AM, meulia.1-at-osu.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 } ----------------------------------------------------------------------- } ----- } } We have been doing our pre-embedding immunogold localizations } incubations for animal tissue in PBS, 0.2% BSA and 10mM Na Azide. } } I am working now on some em pre-embedding localizations on plant } tissue, for which light microscopy localizations have been done using } MTSB buffer (50mM PIPES, 5mM MgSO4, 5mM EGTA, pH~7.0). Is there any } reason for which I should not be using this same MTSB buffer for the } em work? } } Thank you very much for your suggestions. They will definitely save } me trial time. } } Thanks. } } Tea } } } } -- } *************************************** } Tea Meulia, PhD } Director } Molecular and Cellular Imaging Center } Ohio State University/OARDC } 1680 Madison Ave. } Wooster OH 44691 } } tel.: 330-263-3836 or -3828 } fax: 330-202-3563 } } http://www.oardc.ohio-state.edu/mcic } } ***************************************** } } ==============================Original } Headers============================== } 11, 18 -- From meulia.1-at-osu.edu Tue May 16 13:17:06 2006 } 11, 18 -- Received: from defang9.net.ohio-state.edu } (defang9.net.ohio-state.edu [128.146.216.78]) } 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k4GIH5xS001081 } 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 13:17:05 } -0500 } 11, 18 -- Received: from [164.107.85.164] } (dhcp-107-85-164.oardc.ohio-state.edu [164.107.85.164]) } 11, 18 -- by defang9.net.ohio-state.edu (8.13.1/8.13.1) with ESMTP id } k4GIH5AC031731 } 11, 18 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 14:17:05 } -0400 } 11, 18 -- Mime-Version: 1.0 } 11, 18 -- X-Sender: meulia.1-at-pop.service.ohio-state.edu } 11, 18 -- Message-Id: {p05200f03c08fc56f761a-at-[164.107.85.164]} } 11, 18 -- Date: Tue, 16 May 2006 14:17:12 -0400 } 11, 18 -- To: Microscopy Listserver {microscopy-at-microscopy.com} } 11, 18 -- From: Tea Meulia {meulia.1-at-osu.edu} } 11, 18 -- Subject: em immunolocalizations } 11, 18 -- Content-Type: text/plain; charset="us-ascii" ; } format="flowed" } 11, 18 -- X-Spam-Score: undef - spam scanning disabled } 11, 18 -- X-CanItPRO-Stream: outbound } 11, 18 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on } 128.146.216.12 } ==============================End of - } Headers============================== }
==============================Original Headers============================== 7, 22 -- From leunissen-at-aurion.nl Tue May 16 19:23:02 2006 7, 22 -- Received: from mailhub1.otago.ac.nz (mailhub1.otago.ac.nz [139.80.64.218]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4H0N0Hw009205 7, 22 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 19:23:02 -0500 7, 22 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 7, 22 -- by mailhub1.otago.ac.nz (8.13.6/8.13.6) with ESMTP id k4H0MvS7004469; 7, 22 -- Wed, 17 May 2006 12:22:58 +1200 7, 22 -- Received: from ou034063.otago.ac.nz ([139.80.34.63]) 7, 22 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 7, 22 -- id 1Fg9oW-0001BU-P9; Wed, 17 May 2006 12:22:56 +1200 7, 22 -- In-Reply-To: {200605161817.k4GIHShJ001598-at-ns.microscopy.com} 7, 22 -- References: {200605161817.k4GIHShJ001598-at-ns.microscopy.com} 7, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 7, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 22 -- Message-Id: {af5cfb1e829a31cdaf5d162318f247f3-at-aurion.nl} 7, 22 -- Content-Transfer-Encoding: 7bit 7, 22 -- Cc: microscopy-at-microscopy.com 7, 22 -- From: Jan Leunissen {leunissen-at-aurion.nl} 7, 22 -- Subject: Re: [Microscopy] em immunolocalizations 7, 22 -- Date: Wed, 17 May 2006 12:22:57 +1200 7, 22 -- To: meulia.1-at-osu.edu 7, 22 -- X-Mailer: Apple Mail (2.624) ==============================End of - Headers==============================
This should be another interesting AFM experiment, done with liquid cell. It would be helpful, however, to get them to adhere to a substrate first, so that they don't roll around. Contact me off-line and we can probably put together a protocol for you. Also, if you can get samples, we can try to run them here. We have an AFM that can run in liquid.
Hope this is helpful,
Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through 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 03:35 PM 5/16/2006, edelmare-at-muohio.edu wrote:
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==============================Original Headers============================== 18, 17 -- From bfoster-at-mme1.com Tue May 16 21:27:49 2006 18, 17 -- Received: from smtp103.sbc.mail.mud.yahoo.com (smtp103.sbc.mail.mud.yahoo.com [68.142.198.202]) 18, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4H2RnQ9021264 18, 17 -- for {microscopy-at-microscopy.com} ; Tue, 16 May 2006 21:27:49 -0500 18, 17 -- Received: (qmail 16839 invoked from network); 17 May 2006 02:27:47 -0000 18, 17 -- Received: from unknown (HELO barbsd505.mme1.com) (bfostermme-at-sbcglobal.net-at-68.94.44.10 with login) 18, 17 -- by smtp103.sbc.mail.mud.yahoo.com with SMTP; 17 May 2006 02:27:47 -0000 18, 17 -- Message-Id: {7.0.1.0.0.20060516212508.0206a450-at-mme1.com} 18, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 18, 17 -- Date: Tue, 16 May 2006 21:27:55 -0500 18, 17 -- To: edelmare-at-muohio.edu, microscopy Listserver {microscopy-at-microscopy.com} 18, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 18, 17 -- Subject: Re: [Microscopy] Sizing 60-atom bucky balls 18, 17 -- In-Reply-To: {200605162035.k4GKZYsq025773-at-ns.microscopy.com} 18, 17 -- References: {200605162035.k4GKZYsq025773-at-ns.microscopy.com} 18, 17 -- Mime-Version: 1.0 18, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (Dstekl-at-frontiernet.net) from on Tuesday, May 16, 2006 at 19:19:11 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both Dstekl-at-frontiernet.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: Dstekl-at-frontiernet.net Name: Dave Steklenski
Organization: Catherine McAuley School, Rochester NY
Education: K-8 Grade Grammar School
Location: Rochester NY
Question: We are planning an elementary microscopy lab using a variety of natural and synthetic fibers. I would like to locat some photomicrographs of sample fibers so the kids could compare their observations to know appearance samples. I have been unable to find a good set of micrographs on the web.......are there any common references that might have some useable pictures.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (milindphd-at-gmail.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, May 17, 2006 at 00:52:35 ---------------------------------------------------------------------------
Email: milindphd-at-gmail.com Name: Milind Redkar
Organization: University institute of chemical technology
Education: Graduate College
Location: mumbai,India
Question: please let me know, why multilamellar vesicle show maltese crosses when veiwed with crosspolarizing microscopy.?
what is the difference between conoscopy and microscopy with resoect to multilamellar vesicles?
I have a small box of new JEOL 35C electronic parts. Potentiometers, lamps, IC's, transistors,push-button switches, etc. I'll send it to anyone that wants it. Email me at directly.
Owen
Owen P. Mills Director, Materials Characterization & Fabrication Facilities Electron Optics Engineer, Applied Chemical & Morphological Analysis Laboratory
Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
==============================Original Headers============================== 8, 31 -- From opmills-at-mtu.edu Wed May 17 08:10:59 2006 8, 31 -- Received: from node7.mtu.edu (node7.mtu.edu [141.219.69.7]) 8, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HDAxhK028058 8, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 08:10:59 -0500 8, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 8, 31 -- by node7.mtu.edu (8.13.1/8.13.1) with ESMTP id k4HDAwQA028733 8, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:10:58 -0400 8, 31 -- Received: from node2.edge.dcsint.mtu.edu (node2.mtu.edu [141.219.69.2]) 8, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k4HDAwd8013241 8, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:10:58 -0400 8, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) 8, 31 -- by node2.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k4HDAv7g012056 8, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:10:58 -0400 8, 31 -- (envelope-from opmills-at-mtu.edu) 8, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 8, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k4HDAvBG010647 8, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:10:57 -0400 (EDT) 8, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 8, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k4HDAvkp013219 8, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:10:57 -0400 8, 31 -- Mime-Version: 1.0 (Apple Message framework v749.3) 8, 31 -- Content-Transfer-Encoding: 7bit 8, 31 -- Message-Id: {6444BECD-22E3-48E5-9098-8B73F0C2589E-at-mtu.edu} 8, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 8, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 8, 31 -- Subject: free 35C parts 8, 31 -- Date: Wed, 17 May 2006 09:10:55 -0400 8, 31 -- X-Mailer: Apple Mail (2.749.3) 8, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.5.17.55109 8, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
I have 2 Jeol M3 cathodes I'll give anyone that wants them. They were bought for a 100CX TEM. Both are new, but have been on a shelf for 20yrs. Let me know if you want them.
Owen
==============================Original Headers============================== 2, 31 -- From opmills-at-mtu.edu Wed May 17 08:15:42 2006 2, 31 -- Received: from node7.mtu.edu (node7.mtu.edu [141.219.69.7]) 2, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HDFfxX001183 2, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 08:15:41 -0500 2, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 2, 31 -- by node7.mtu.edu (8.13.1/8.13.1) with ESMTP id k4HDFfhr029034 2, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:15:41 -0400 2, 31 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 2, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k4HDFfs3005269 2, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:15:41 -0400 2, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) 2, 31 -- by node34.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k4HDFeF4020104 2, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:15:40 -0400 2, 31 -- (envelope-from opmills-at-mtu.edu) 2, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 2, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k4HDFeVh014345 2, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:15:40 -0400 (EDT) 2, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 2, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k4HDFdDw005259 2, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:15:39 -0400 2, 31 -- Mime-Version: 1.0 (Apple Message framework v749.3) 2, 31 -- Content-Transfer-Encoding: 7bit 2, 31 -- Message-Id: {E872D0DE-5441-4783-AD47-788034ED7801-at-mtu.edu} 2, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 2, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 2, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 2, 31 -- Subject: Free JEOL Denka LaB6 cathodes 2, 31 -- Date: Wed, 17 May 2006 09:15:38 -0400 2, 31 -- X-Mailer: Apple Mail (2.749.3) 2, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.5.17.55109 2, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
Hello TEM community: I have LEO 912AB TEM and need to order a new LaB6 filament. I can't for the life of me remember or find in my files what I ordered the last time except that it was a Denka. If anyone has this information and would pass it on I would be truly grateful. bob harris
Guelph Regional Imaging Facility Dept.of Molecular and Cellular Biology New Science Complex 488 Gordon St. Univ of Guelph Guelph,On, Canada N1G 2W1 ph: 519-824-4120 ext. 56409/58962 Fax: 519-837-1802
==============================Original Headers============================== 2, 27 -- From bharris-at-uoguelph.ca Wed May 17 08:39:32 2006 2, 27 -- Received: from mailhub.cs.uoguelph.ca (mailhub.cs.uoguelph.ca [131.104.94.205]) 2, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HDdWnD015600 2, 27 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 08:39:32 -0500 2, 27 -- Received: from pippin.cs.uoguelph.ca ([131.104.93.20]) 2, 27 -- by mailhub.cs.uoguelph.ca (8.13.1/8.13.1) with ESMTP id k4HDdVX5007779 2, 27 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:39:31 -0400 2, 27 -- Received: from pippin.cs.uoguelph.ca (localhost.localdomain [127.0.0.1]) 2, 27 -- by pippin.cs.uoguelph.ca (8.12.11/8.12.11) with ESMTP id k4HDdVlS018349 2, 27 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:39:31 -0400 2, 27 -- Received: (from nobody-at-localhost) 2, 27 -- by pippin.cs.uoguelph.ca (8.12.11/8.12.11/Submit) id k4HDdVGU018347 2, 27 -- for microscopy-at-microscopy.com; Wed, 17 May 2006 09:39:31 -0400 2, 27 -- X-Authentication-Warning: pippin.cs.uoguelph.ca: nobody set sender to bharris-at-uoguelph.ca using -f 2, 27 -- Received: from 131.104.80.123 ([131.104.80.123]) 2, 27 -- by webmail.uoguelph.ca (IMP) with HTTP 2, 27 -- for {bharris-at-staff.mail.uoguelph.ca} ; Wed, 17 May 2006 09:39:31 -0400 2, 27 -- Message-ID: {1147873171.446b2793c3a31-at-webmail.uoguelph.ca} 2, 27 -- Date: Wed, 17 May 2006 09:39:31 -0400 2, 27 -- From: Bob Harris {bharris-at-uoguelph.ca} 2, 27 -- To: MSA listserver {microscopy-at-microscopy.com} 2, 27 -- Subject: LaB6 filament for a LEO 912AB 2, 27 -- MIME-Version: 1.0 2, 27 -- Content-Type: text/plain; charset=ISO-8859-1 2, 27 -- Content-Transfer-Encoding: 8bit 2, 27 -- User-Agent: Internet Messaging Program (IMP) 3.2.4 2, 27 -- X-Scanned-By: MIMEDefang 2.52 on 131.104.94.205 ==============================End of - Headers==============================
Have a look at http://www.microscopy-uk.org.uk/index.html under MENU
It's a bit of struggle with all the links, but under Best Image Galleries & Collections have a look at the SEM (scanning electron microscope) - the first one. They aren't light microscopy but SEMs are great for fibre morphology surface details.
I found things like Cellulose fibers (fibres) in toilet paper, paper towels, T shirt cotton with dirt, asbestos (best to view that on-line), woven silk etc.. [In 'Dennis Kunkel Microscopy, Inc.' link.]
Also search through the microscopy primer site, they have loads of stock images
I found rabbit hair, silk, nylon (all under polarised light microscopy so they appear brightly coloured - rather unlike standard microscope images).
The microscopy primer also tells you loads about microscopes (and you can operate them virtually).
Plus try the hobby site http://www.btinternet.com/~stephen.durr/ as it has links that may provide something (but most sites are interested in things like plants, animals and pond life).
You may be able to get a library loan of an old book that has suitable pictures for scanning, e.g.
Textile fiber atlas : A collection of photomicrographs of old and new textile fibers, by Werner Von Bergen (Jan 1, 1949).
The microscopy of animal textile fibres,: Including methods for the complete analysis of fibre blends. 235 half-tone and ll colour photomicrographs, 88 line drawings by Alec Blakey Wildman (Jan 1, 1954)
I found the book links on amazon.com (both are out of print).
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: Dstekl-at-frontiernet.net [mailto:Dstekl-at-frontiernet.net] Sent: 17 May 2006 13:03 To: keith.morris-at-ucl.ac.uk
This Question was submitted to Ask-A-Microscopist by (Dstekl-at-frontiernet.net) from on Tuesday, May 16, 2006 at 19:19:11 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both Dstekl-at-frontiernet.net as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: Dstekl-at-frontiernet.net Name: Dave Steklenski
Organization: Catherine McAuley School, Rochester NY
Education: K-8 Grade Grammar School
Location: Rochester NY
Question: We are planning an elementary microscopy lab using a variety of natural and synthetic fibers. I would like to locat some photomicrographs of sample fibers so the kids could compare their observations to know appearance samples. I have been unable to find a good set of micrographs on the web.......are there any common references that might have some useable pictures.
THANK you for the help!!!
==============================Original Headers============================== 31, 26 -- From keith.morris-at-ucl.ac.uk Wed May 17 09:12:43 2006 31, 26 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 31, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HEChiI025888 31, 26 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:12:43 -0500 31, 26 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 31, 26 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 31, 26 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 31, 26 -- id 1FgMlV-00042h-2D; Wed, 17 May 2006 15:12:41 +0100 31, 26 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 31, 26 -- To: {Dstekl-at-frontiernet.net} 31, 26 -- Cc: {Microscopy-at-microscopy.com} 31, 26 -- Subject: RE: [Microscopy] [Filtered] AskAMicroscopist: elementary microscopy lab 31, 26 -- Date: Wed, 17 May 2006 15:11:46 +0100 31, 26 -- Message-ID: {000601c679bb$d5356790$7b865290-at-keithhigrade} 31, 26 -- MIME-Version: 1.0 31, 26 -- Content-Type: text/plain; 31, 26 -- charset="us-ascii" 31, 26 -- Content-Transfer-Encoding: 7bit 31, 26 -- X-Mailer: Microsoft Office Outlook 11 31, 26 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 31, 26 -- Thread-Index: AcZ5qdZktnXyK/AgTe+ppRP1nSzVngACzwsA 31, 26 -- In-Reply-To: {200605171202.k4HC2nPb021701-at-ns.microscopy.com} 31, 26 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 31, 26 -- X-UCL-MailScanner: Found to be clean 31, 26 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 31, 26 -- X-Spam-Status: No ==============================End of - Headers==============================
Ryan, In addition to all the other suggestions about small sample size, out of the lens field, etc., also try running your sample through a degausser just before putting it in the SEM. Any residual magnetism is going to adversely affect a high mag image. I can remember (too many years ago) swearing at the SEM I operated (and didn't particularly like) because my resolution was terrible, then remembering that my sample was a piece of carbon steel. After degaussing, the resolution was fine.
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: lewryan-at-gmail.com [mailto:lewryan-at-gmail.com] Sent: Monday, May 15, 2006 6:07 PM To: kenconverse-at-qualityimages.biz
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Email: lewryan-at-gmail.com Name: Ryan
Organization: Dalhousie University
Title-Subject: [Filtered] SEM imaging on sample with magnetic substrate
Question: I have a saple with deposits of an alloy of Sn-Co-C on a magnetic stainless steel substrate (430 stainless steel). I'm using a cold field emission microscope and am having trouble getting high resolution images. Could you suggest what setting I should use to get started?
==============================Original Headers============================== 6, 12 -- From zaluzec-at-microscopy.com Mon May 15 17:03:45 2006 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4FM3ipN026657 6, 12 -- for {microscopy-at-microscopy.com} ; Mon, 15 May 2006 17:03:44 -0500 6, 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- Message-Id: {p06110401c08eab2f530f-at-[206.69.208.22]} 6, 12 -- Date: Mon, 15 May 2006 17:03:44 -0500 6, 12 -- To: microscopy-at-microscopy.com 6, 12 -- From: lewryan-at-gmail.com (by way of MicroscopyListserver) 6, 12 -- Subject: viaWWW: SEM imaging on sample with magnetic substrate 6, 12 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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==============================Original Headers============================== 23, 30 -- From kenconverse-at-qualityimages.biz Wed May 17 09:45:19 2006 23, 30 -- Received: from dpmailmta01.doteasy.com (dpmailmta01.doteasy.com [65.61.209.91]) 23, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HEjJYC003869 23, 30 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 17 May 2006 09:45:19 -0500 23, 30 -- Received: from qualityimages.biz (unverified [192.168.101.16]) 23, 30 -- by dpmta01.doteasy.com (DEO) with ESMTP id 4857839 23, 30 -- for multiple; Wed, 17 May 2006 07:58:13 -0700 23, 30 -- Received: from Ken [24.53.5.245] by qualityimages.biz with ESMTP 23, 30 -- (SMTPD32-8.05) id A6F25693008A; Wed, 17 May 2006 07:45:06 -0700 23, 30 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 23, 30 -- To: {lewryan-at-gmail.com} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 23, 30 -- Subject: RE: [Microscopy] viaWWW: SEM imaging on sample with magnetic substrate 23, 30 -- Date: Wed, 17 May 2006 10:46:18 -0400 23, 30 -- Message-ID: {005101c679c0$ac184030$6501a8c0-at-Ken} 23, 30 -- MIME-Version: 1.0 23, 30 -- Content-Type: text/plain; 23, 30 -- charset="us-ascii" 23, 30 -- X-Priority: 3 (Normal) 23, 30 -- X-MSMail-Priority: Normal 23, 30 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 23, 30 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 23, 30 -- Importance: Normal 23, 30 -- In-Reply-To: {200605152206.k4FM6uej031789-at-ns.microscopy.com} 23, 30 -- X-IMSTrailer: __IMail_7__ 23, 30 -- X-Server: High Performance Mail Server - http://surgemail.com r=34189668 23, 30 -- X-NotAscii: charset=us-ascii 23, 30 -- X-IP-stats: Incoming Last 0, First 22, in=226356, out=0, spam=0 23, 30 -- X-External-IP: 192.168.101.16 23, 30 -- Content-Transfer-Encoding: 8bit 23, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4HEjJYC003869 ==============================End of - Headers==============================
I forwarded your request to Scott Stoeffler, a member of our Optical Microscopy group, and he provided these suggestions:
Any good basic book on textile science will have fiber photomicrographs. Margery Joseph's Introductory Textile Science is a good one and available pretty cheaply.
There is also a CD available with a lot of fiber pictures, but it's pretty expensive:
http://www.atexinc.com/digital_textiles_(cd).htm
Natural fibers such as cotton, linen, wool and silk are easier to distinguish based on morphology alone. Without polarized light, synthetics aren't easily distinguishable, although you can look at cross-sections.
You might also take a look at our on-line Atlas of Microscopic Particles at www.mccroneatlas.com. If you do a basic search on fibers, you'll get some examples of cloth and paper fiber images with some background information.
Hope this helps.
Regards,
Elaine
********************************************************************* Elaine F.Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited. *********************************************************************
-----Original Message----- X-from: Dstekl-at-frontiernet.net [mailto:Dstekl-at-frontiernet.net] Sent: Wednesday, May 17, 2006 6:55 AM To: Elaine F. Schumacher
This Question was submitted to Ask-A-Microscopist by (Dstekl-at-frontiernet.net) from on Tuesday, May 16, 2006 at 19:19:11 Remember to consider the Grade/Age of the student when considering the Question ------------------------------------------------------------------------ --- Please reply to both Dstekl-at-frontiernet.net as well as to the Microscopy Listserver ------------------------------------------------------------------------ ---
Email: Dstekl-at-frontiernet.net Name: Dave Steklenski
Organization: Catherine McAuley School, Rochester NY
Education: K-8 Grade Grammar School
Location: Rochester NY
Question: We are planning an elementary microscopy lab using a variety of natural and synthetic fibers. I would like to locat some photomicrographs of sample fibers so the kids could compare their observations to know appearance samples. I have been unable to find a good set of micrographs on the web.......are there any common references that might have some useable pictures.
The standard configuration of Denka cathode for Leo Microscopes is the M3-CA, which has a 15 micron round tip. For greater brightness, especially in TEM applications, many users prefer the M3-CA sharp 60/10 or M3-CA sharp 60/5. Please feel free to contact me off line if you would like to discuss the differences.
Sincerely, Mike Nesta
Michael R. Nesta Managing Director Energy Beam Sciences, Inc. Tel: 860 653-0411 Fax: 860 653-0422 MNesta-at-ebsciences.com www.ebsciences.com “ADDING BRILLIANCE TO YOUR VISION”
bharris-at-uoguelph.ca wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello TEM community: I have LEO 912AB TEM and need to order a new LaB6 } filament. I can't for the life of me remember or find in my files what I } ordered the last time except that it was a Denka. If anyone has this } information and would pass it on I would be truly grateful. bob harris } } Guelph Regional Imaging Facility } Dept.of Molecular and Cellular } Biology } New Science Complex } 488 Gordon St. } Univ of Guelph } Guelph,On, Canada } N1G 2W1 } ph: 519-824-4120 ext. 56409/58962 } Fax: 519-837-1802 } } ==============================Original Headers============================== } 2, 27 -- From bharris-at-uoguelph.ca Wed May 17 08:39:32 2006 } 2, 27 -- Received: from mailhub.cs.uoguelph.ca (mailhub.cs.uoguelph.ca [131.104.94.205]) } 2, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HDdWnD015600 } 2, 27 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 08:39:32 -0500 } 2, 27 -- Received: from pippin.cs.uoguelph.ca ([131.104.93.20]) } 2, 27 -- by mailhub.cs.uoguelph.ca (8.13.1/8.13.1) with ESMTP id k4HDdVX5007779 } 2, 27 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:39:31 -0400 } 2, 27 -- Received: from pippin.cs.uoguelph.ca (localhost.localdomain [127.0.0.1]) } 2, 27 -- by pippin.cs.uoguelph.ca (8.12.11/8.12.11) with ESMTP id k4HDdVlS018349 } 2, 27 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 09:39:31 -0400 } 2, 27 -- Received: (from nobody-at-localhost) } 2, 27 -- by pippin.cs.uoguelph.ca (8.12.11/8.12.11/Submit) id k4HDdVGU018347 } 2, 27 -- for microscopy-at-microscopy.com; Wed, 17 May 2006 09:39:31 -0400 } 2, 27 -- X-Authentication-Warning: pippin.cs.uoguelph.ca: nobody set sender to bharris-at-uoguelph.ca using -f } 2, 27 -- Received: from 131.104.80.123 ([131.104.80.123]) } 2, 27 -- by webmail.uoguelph.ca (IMP) with HTTP } 2, 27 -- for {bharris-at-staff.mail.uoguelph.ca} ; Wed, 17 May 2006 09:39:31 -0400 } 2, 27 -- Message-ID: {1147873171.446b2793c3a31-at-webmail.uoguelph.ca} } 2, 27 -- Date: Wed, 17 May 2006 09:39:31 -0400 } 2, 27 -- From: Bob Harris {bharris-at-uoguelph.ca} } 2, 27 -- To: MSA listserver {microscopy-at-microscopy.com} } 2, 27 -- Subject: LaB6 filament for a LEO 912AB } 2, 27 -- MIME-Version: 1.0 } 2, 27 -- Content-Type: text/plain; charset=ISO-8859-1 } 2, 27 -- Content-Transfer-Encoding: 8bit } 2, 27 -- User-Agent: Internet Messaging Program (IMP) 3.2.4 } 2, 27 -- X-Scanned-By: MIMEDefang 2.52 on 131.104.94.205 } ==============================End of - Headers============================== }
==============================Original Headers============================== 9, 28 -- From mnesta-at-ebsciences.com Wed May 17 10:33:21 2006 9, 28 -- Received: from ylpvm43.prodigy.net (ylpvm43-ext.prodigy.net [207.115.57.74]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HFXKhp024690 9, 28 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 10:33:21 -0500 9, 28 -- Received: from pimout6-ext.prodigy.net (pimout6-int.prodigy.net [207.115.4.22]) 9, 28 -- by ylpvm43.prodigy.net (8.12.10 outbound/8.12.10) with ESMTP id k4HFXMWH030373 9, 28 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 11:33:22 -0400 9, 28 -- X-ORBL: [69.182.224.2] 9, 28 -- Received: from mail.ebsciences.com ([69.182.224.2]) 9, 28 -- by pimout6-ext.prodigy.net (8.13.6 out.dk/8.13.6) with ESMTP id k4HFXJvT180922; 9, 28 -- Wed, 17 May 2006 11:33:19 -0400 9, 28 -- Received: from dhcp-206-53-66-197.myeastern.com ([206.53.66.197] helo=[192.168.0.15]) 9, 28 -- by mail.ebsciences.com with esmtpsa (TLSv1:AES256-SHA:256) 9, 28 -- (Exim 4.54) 9, 28 -- id 1FgO1W-0005sO-RY; Wed, 17 May 2006 11:33:19 -0400 9, 28 -- Message-ID: {446B4245.7050903-at-ebsciences.com} 9, 28 -- Date: Wed, 17 May 2006 11:33:25 -0400 9, 28 -- From: Mike Nesta {mnesta-at-ebsciences.com} 9, 28 -- Organization: Energy Beam Sciences 9, 28 -- User-Agent: Thunderbird 1.5.0.2 (Windows/20060308) 9, 28 -- MIME-Version: 1.0 9, 28 -- To: bharris-at-uoguelph.ca 9, 28 -- CC: microscopy-at-microscopy.com 9, 28 -- Subject: Re: [Microscopy] LaB6 filament for a LEO 912AB 9, 28 -- References: {200605171342.k4HDgb4g020997-at-ns.microscopy.com} 9, 28 -- In-Reply-To: {200605171342.k4HDgb4g020997-at-ns.microscopy.com} 9, 28 -- Content-Type: text/plain; charset=windows-1252; format=flowed 9, 28 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
A question to whose who have made some recent tests on FEG-SEM.
I would have advices about the differences which can be practically seen between the Zeiss Supra 40 et the Supra 55. Resolutions annonced are quite different. I want to know if it's only a spec difference, or if practically it's easy to see it. We have made tests on the 55, but the budget doesn't follow... All other advices are welcome.
Thanks
-- 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
Seeing at the Nanoscale IV, July 17-20, University of Pennsylvania, Philadelphia.
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Someone had said that distilled water will cause copper tubing to ionize, and correction will be faster than tap water. Has anybody heard this?
Ann Fook Yang EM Unit/ Unite EM AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 yanga-at-agr.gc.ca Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu] Sent: Tuesday, May 16, 2006 6:50 PM To: Yang, Ann-Fook
Just wanted to thank everyone for the responses to my posting about the TEM water chiller additives. I'll try just using distilled water and monitoring the system more frequently. Thank you! 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)
The only additive I use is colloidal silver, sold as a health food item by Natural Immunogenics 888-328-8840. This method, suggested to me by Vitaly Feingold has worked very well. I also add a quart or so of tap water to the DI water so it is not so aggressive on the copper.
I have no connection with the silver supplier.
David Rothbard
beth-at-plantbio.uga.edu wrote:
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==============================Original Headers============================== 8, 20 -- From rothbardd-at-netscape.net Wed May 17 11:14:37 2006 8, 20 -- Received: from imo-d01.mx.aol.com (imo-d01.mx.aol.com [205.188.157.33]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HGEb2W032643 8, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 11:14:37 -0500 8, 20 -- Received: from rothbardd-at-netscape.net 8, 20 -- by imo-d01.mx.aol.com (mail_out_v38_r7.5.) id w.1b1.11dc389c (16239) 8, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 17 May 2006 12:14:28 -0400 (EDT) 8, 20 -- Received: from netscape.net (mow-d20.webmail.aol.com [205.188.139.161]) by air-in03.mx.aol.com (v109.12) with ESMTP id MAILININ33-3f6f446b4be43d1; Wed, 17 May 2006 12:14:28 -0400 8, 20 -- Date: Wed, 17 May 2006 12:14:28 -0400 8, 20 -- From: rothbardd-at-netscape.net 8, 20 -- To: Microscopy-at-microscopy.com 8, 20 -- Subject: RE: TEM water recirculator 8, 20 -- MIME-Version: 1.0 8, 20 -- Message-ID: {30E4C578.63394E02.034D9A6A-at-netscape.net} 8, 20 -- X-Mailer: Atlas Mailer 2.0 8, 20 -- X-AOL-IP: 199.196.144.13 8, 20 -- X-AOL-Language: english 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit 8, 20 -- X-Spam-Flag: NO ==============================End of - Headers==============================
Distilled water is fine. It is the deionized water that cause problems to the copper tubing due to the lack of ions in it.
Zhaojie Zhang Microscopy Core Facility University of Wyoming Laramie, WY 82071
-----Original Message----- X-from: YANGA-at-AGR.GC.CA [mailto:YANGA-at-AGR.GC.CA] Sent: Wednesday, May 17, 2006 8:47 AM To: Z.J. Zhang
Hi Beth,
Someone had said that distilled water will cause copper tubing to ionize, and correction will be faster than tap water. Has anybody heard this?
Ann Fook Yang EM Unit/ Unite EM AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 yanga-at-agr.gc.ca Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu] Sent: Tuesday, May 16, 2006 6:50 PM To: Yang, Ann-Fook
Just wanted to thank everyone for the responses to my posting about the TEM water chiller additives. I'll try just using distilled water and monitoring the system more frequently. Thank you! 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)
That is a difficult situation. First off, the Supra 55 was discontinued for some unknown reason, leaving the Supra 55VP still viable. IMO, the 55 is superior to the 55VP. Why the 40 has lower resolution than the 55 is unknown to me. At 200KX-350KX, I rather doubt that one could tell the difference in resolution between the 40 and 55VP. But one probably could see a difference between the 40 and 55. this assumes identical conditions (low current, 30u center aperture, 3mm WD, 20KV, in-lens detector).
gary g.
At 08:37 AM 5/17/2006, you wrote:
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The questions you have asked require a whole day lecture on polarized light, but here are the fundamentals.
Materials that respond to polarized light have different refractive indices along different axes. These axes are always perpendicular to each other, but not necessarily to the crystal edges or material directions (although they typically lie along the long and short edges of a fiber and along the direction of draw and perpendicular in a film.. but more about that later). The refractive index is a measurement of the interaction of the electric field in light with the electric field in matter: the greater the interaction, the higher the value. Mathematically, you can calculate RI by dividing the velocity of light in vacuum (300,000 km/s) with the velocity of light in the material.
I can't send diagrams in this email (will do so, if you contact me off line), but imagine a rectangle as representing your object, with the short side having lower RI and the long side having higher RI (Step 1) . (This is highly simplified, but works). Next, imagine the crossed polars, with direction of vibration transmitted through the first polar as vibrating East-West and the permitted direction of vibration through the second polar (the "analyzer") as North-South (Step 2).
Step 3: Cross the polars and put the object in between. Imagine that you are looking down on this "sandwich". a. When either the short side or long side of the rectangle is parallel to the polarizer, only that direction receives energy from the polarized light. Since that light is vibrating E-W, it will not pass the analyzer. Rather, it will be absorbed, so the crystal appears dark in these two orientations. b. Imagine rotating the crystal into any 45 degree position. In this orientation, both RI directions receive energy from the polarizer and, if you complete the rectangle so that you have a resultant vector, you will see that the vector has components that will pass through the analyzer. In this position, the crystal will appear bright between crossed polars.
Now for your multi-lamellar vesicle. The fact that it has areas that appear bright indicates that its internal structure generates differences in electrical field with direction. In other words, it is anisotropic. If you consider how the lamellae are laid down in the vesicle, this makes perfect sense. The maltese cross that you are seeing indicate the two directions of the primary RI's. In each of these positions, only one RI is visible to the incoming polarized light; the same situation as described above in which the short or long side of the crystal is parallel to the direction of the polarizer or analyzer.
Actually, if you removed the analyzer and did RI tests with just the polarizer in place, then rotated the polarizer 90 degrees and did another RI test, you could actually determine the RI in each of those directions.
I don't have direct experience with this type of vesicle, but I would assume that, like starch and polymer spherulites, if you rotate your sample between XPol, the cross will stay in the same orientation indicating that your lamellae are sitting radially within the vesicle.
Hope this was helpful.
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through 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 06:59 AM 5/17/2006, milindphd-at-gmail.com wrote:
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==============================Original Headers============================== 22, 17 -- From bfoster-at-mme1.com Wed May 17 11:22:05 2006 22, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 22, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4HGM5Yq014511 22, 17 -- for {microscopy-at-microscopy.com} ; Wed, 17 May 2006 11:22:05 -0500 22, 17 -- Received: (qmail 5501 invoked by uid 2020); 17 May 2006 11:38:14 -0500 22, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 22, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 17 May 2006 11:38:14 -0500 22, 17 -- Message-Id: {7.0.1.0.0.20060517105231.020eaa40-at-mme1.com} 22, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 22, 17 -- Date: Wed, 17 May 2006 11:22:11 -0500 22, 17 -- To: milindphd-at-gmail.com, microscopy Listserver {microscopy-at-microscopy.com} 22, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 22, 17 -- Subject: Re: [Microscopy] AskAMicroscopist: multilamellar vesicle 22, 17 -- In-Reply-To: {200605171159.k4HBx9Nh015503-at-ns.microscopy.com} 22, 17 -- References: {200605171159.k4HBx9Nh015503-at-ns.microscopy.com} 22, 17 -- Mime-Version: 1.0 22, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Yes, this can be the case. Distilled water can often be more corrosive to piping than regular tap water, it does depend upon the water chemistry of the tap water. What you really want is de-oxygenated water.
Beth, if you'd like more info, I can send you a synopsis of a recent discussion on this subject from a while back. Or if you prefer, you may email me directly and we could talk more about this, as you wish.
dj
On Wed, 17 May 2006 YANGA-at-AGR.GC.CA wrote:
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I am looking for an older Oxford Pulse Processor so I can do some testing of EDX detectors. The model number can go back to #2020 and forward. I don't need the complete analyzer system. If anyone has one of these pulse processors that is available for sale, please contact me directly at my email below. Kind Regards, Jim Nicolino
PulseTor/AAT 1816 St. Johns Bluff Road Suite 305 Jacksonville, Florida 32246 904.646.3069 FAX 904.646.3131 JNicolino-at-comcast.net
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As the President, Officers, and only member of the Half-Norwegian (on my Mother's side) Electron Microscopy Society, I would like to wish all you listers a Happy Norwegian Independence Day!!
Also, thank you for all your recent help on nano-particles in agarose and the fascinating string on ethics.
Uff Da!
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
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On May 17, 2006, at 8:38 AM, YANGA-at-AGR.GC.CA wrote:
} Someone had said that distilled water will cause copper tubing to } ionize, and correction will be faster than tap water. Has anybody } heard this? } Dear Ann Fook, I also have heard this. I have not done the experiment, but if [Cu+] = [Cu++] = 0, then any dissolved oxidizing agent will cause Cu to be ionized, so there is good reason to believe that distilled H2O will be more corrosive. I would not advise using it in an EM. 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
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On May 17, 2006, at 9:20 AM, ZZhang-at-uwyo.edu wrote:
} Distilled water is fine. It is the deionized water that cause problems } to the copper tubing due to the lack of ions in it.
Dear Zhaojie, Distillation will remove non-volatile ions, so unless the tap water has something like NH4HCO3 in it, it will not have many ions after distillation. 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
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This Question was submitted to Ask-A-Microscopist by (Mayya_Saab-at-dps-consulting.com) from http://microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, May 17, 2006 at 13:52:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both Mayya_Saab-at-dps-consulting.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: Mayya_Saab-at-dps-consulting.com Name: Mayya Saab
Education: Undergraduate College
Location: Herndon, VA, Fairfax
Title: darkfield microscopy
Question: What is this field? How can it help an individual?
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Email: xxu-at-ngimat.com Name: Xiao Xu
Organization: nGimat Co
Title-Subject: [Filtered] Replace inner bake heater on Hitachi S-800 SEM
Question: Dear MSA Listserver Colleagues,
Does anyone have experience of replacing the inner bake heater on Hitachi S-800 SEM or recommend an independent contractor to do the job in Atlanta, GA area? Thank you very mcuh!
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Question: I am working on a breaded food and I have been trying to establish protocol for staining my sample to obtain images from Confocal laser microscope. I know I can use FITC for the protein phase, but then the procedure is not really clear. The fat phase staining protocol is still not clear either. Can anybody provide useful information.Thank you
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Email: kempfsc-at-auburn.edu Name: Steve Kempf
Organization: Auburn University
Title-Subject: [Filtered] Running away sections
Question: I've had the strangest thing happen this past weekend and since and no one who I've talked to (some very experienced with thin sectioning) has been able to help.
I had been sectioning a block of tissue embedded in Poly/Bed 812 at 50 nm without difficulty using a Diatome diamond knife on a Leica Ultracut microtome, getting beautiful silver sections, spredding then by wafting a toothpick dipped in chloroform over them, and picking them up on formvar coated slot grids. All has been well until this weekend. Everything was set-up as it had been the previous day. Same knife, same boat water, same toothpicks, same choroform, same block, etc. As was the case previously, I got beautiful silver sections, but when I when to spread them with chloroform on a toothpick, they ran away as I moved the toothpick close to them, lickety-split. I could chase them around the boat with the toothpick. When I tried to pick them up on a grid, I couldn't. They would just slide off, back onto the water surface in the boat. Arrrrgh! Naturally, this is happening just as I reach the critical spot in the tissue I'm sectioning.
Not to be deterred, I cleaned the boat by washing it with clean water, got new beakers to hold the water I use, and tried again. The same thing happened. At that point I called it quits for the day and hoped it was some strange environmental effect that would disappear the following week. No Joy! When I sat down to section today, the same thing happened. I tried cleaning the knife boat with 0.2% ethanol in water, got new glassware, new chloroform, new toothpicks, etc. Still no joy?
So, my question is, can anyone give me some guidance as to what might be going on? I'm at my wits end.
Oh, one other thing, if I let the chloroform evaporate off the dipped toothpick, the sections don't run away from it.
It sounds to me like a static electricity problem. Is your air suddenly much drier? i.e. did they turn on the air conditioner? We found many years ago that we could block this with those old darkroom "dustfree" brushes with polonium strips in them. I don't know if those are still available.
Joel
Date sent: Wed, 17 May 2006 19:39:32 -0500 To: jbs-at-temple.edu X-from: kempfsc-at-auburn.edu Send reply to: kempfsc-at-auburn.edu
I would like to test 3D reconstruction from tilted images (jpeg, tiff or bmp) from an Hitachi S4000 SEM. Does anyone knows a software that could do such thing ? It has to be free, because it is just a test (at the moment) and running under MS-Windows. I know the easiest way yo do that is to paint 2 tilted images (with a tilt difference of about 8 degrees) in red and green with a photoshop-style software and use special glasses, but I would like to display something that looks like an AFM map on the screen. If anyone has experienced that type of job, I would be glad to have advices.
Ludovic Pinier
==============================Original Headers============================== 3, 28 -- From ludovic.pinier-at-thalesgroup.com Thu May 18 02:10:14 2006 3, 28 -- Received: from thsmsgxrt12p.thalesgroup.com (thsmsgxrt12p.thalesgroup.com [192.54.144.135]) 3, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4I7AC2X010005 3, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 02:10:14 -0500 3, 28 -- Received: from thsmsgirt22p.corp.thales (unknown [10.33.231.6]) 3, 28 -- by thsmsgxrt12p.thalesgroup.com (Postfix) with ESMTP id 5F29444088 3, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:10:09 +0200 (CEST) 3, 28 -- Received: from thsmsgiav13p.corp.thales (10.33.231.33) by thsmsgirt22p.corp.thales (7.2.055.4) 3, 28 -- id 44447C950051D4FD for Microscopy-at-microscopy.com; Thu, 18 May 2006 09:10:08 +0200 3, 28 -- Received: from (10.33.231.2) by thsmsgiav13p.corp.thales via smtp 3, 28 -- id 4904_57585078_e63d_11da_9ecd_0014221d46b3; 3, 28 -- Thu, 18 May 2006 07:10:08 +0000 3, 28 -- Received: from frontalorsay.fr.trt.thales (unknown [10.33.5.100]) 3, 28 -- by thsmsgirt12p.corp.thales (Postfix) with ESMTP id 529F33C019 3, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:10:09 +0200 (CEST) 3, 28 -- Received: from frontalorsay.fr.trt.thales (52.21.47.10) by frontalorsay.fr.trt.thales (NPlex 6.5.026) 3, 28 -- id 446025F0000134BC for Microscopy-at-microscopy.com; Thu, 18 May 2006 09:17:24 +0200 3, 28 -- Received: from thalesgroup.com ([52.21.23.226]) by frontalorsay with InterScan Messaging Security Suite; Thu, 18 May 2006 09:17:23 +0200 3, 28 -- Message-ID: {446C1DCE.AFD1785B-at-thalesgroup.com} 3, 28 -- Date: Thu, 18 May 2006 09:10:06 +0200 3, 28 -- From: ludovic pinier {ludovic.pinier-at-thalesgroup.com} 3, 28 -- X-Mailer: Mozilla 4.78 [fr] (Windows NT 5.0; U) 3, 28 -- X-Accept-Language: fr 3, 28 -- MIME-Version: 1.0 3, 28 -- To: Microscopy-at-microscopy.com 3, 28 -- Subject: looking for 3D reconstruction software 3, 28 -- Content-Type: text/plain; charset=us-ascii 3, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
For a free program for 3D surface topography have a look at
Measuring Surface Topography with Scanning Electron Microscopy. I. EZEImage: A Program to Obtain 3D Surface Data Ezequiel Ponz,1 Juan Luis Ladaga,2 and Rita Dominga Bonetto1*
which appeared in Microscopy and Microanalysis 12 (2006) 170-177.
Best regards, Jřrgen.
= = = = = = = = = = = = = = = = =
Joergen B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
-----Original Message----- X-from: ludovic.pinier-at-thalesgroup.com [mailto:ludovic.pinier-at-thalesgroup.com] Sent: Thursday, May 18, 2006 9:12 AM To: j.bilde-at-risoe.dk
I would like to test 3D reconstruction from tilted images (jpeg, tiff or bmp) from an Hitachi S4000 SEM. Does anyone knows a software that could do such thing ? It has to be free, because it is just a test (at the moment) and running under MS-Windows. I know the easiest way yo do that is to paint 2 tilted images (with a tilt difference of about 8 degrees) in red and green with a photoshop-style software and use special glasses, but I would like to display something that looks like an AFM map on the screen. If anyone has experienced that type of job, I would be glad to have advices.
Ludovic Pinier
==============================Original Headers============================== 3, 28 -- From ludovic.pinier-at-thalesgroup.com Thu May 18 02:10:14 2006 3, 28 -- Received: from thsmsgxrt12p.thalesgroup.com (thsmsgxrt12p.thalesgroup.com [192.54.144.135]) 3, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4I7AC2X010005 3, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 02:10:14 -0500 3, 28 -- Received: from thsmsgirt22p.corp.thales (unknown [10.33.231.6]) 3, 28 -- by thsmsgxrt12p.thalesgroup.com (Postfix) with ESMTP id 5F29444088 3, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:10:09 +0200 (CEST) 3, 28 -- Received: from thsmsgiav13p.corp.thales (10.33.231.33) by thsmsgirt22p.corp.thales (7.2.055.4) 3, 28 -- id 44447C950051D4FD for Microscopy-at-microscopy.com; Thu, 18 May 2006 09:10:08 +0200 3, 28 -- Received: from (10.33.231.2) by thsmsgiav13p.corp.thales via smtp 3, 28 -- id 4904_57585078_e63d_11da_9ecd_0014221d46b3; 3, 28 -- Thu, 18 May 2006 07:10:08 +0000 3, 28 -- Received: from frontalorsay.fr.trt.thales (unknown [10.33.5.100]) 3, 28 -- by thsmsgirt12p.corp.thales (Postfix) with ESMTP id 529F33C019 3, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:10:09 +0200 (CEST) 3, 28 -- Received: from frontalorsay.fr.trt.thales (52.21.47.10) by frontalorsay.fr.trt.thales (NPlex 6.5.026) 3, 28 -- id 446025F0000134BC for Microscopy-at-microscopy.com; Thu, 18 May 2006 09:17:24 +0200 3, 28 -- Received: from thalesgroup.com ([52.21.23.226]) by frontalorsay with InterScan Messaging Security Suite; Thu, 18 May 2006 09:17:23 +0200 3, 28 -- Message-ID: {446C1DCE.AFD1785B-at-thalesgroup.com} 3, 28 -- Date: Thu, 18 May 2006 09:10:06 +0200 3, 28 -- From: ludovic pinier {ludovic.pinier-at-thalesgroup.com} 3, 28 -- X-Mailer: Mozilla 4.78 [fr] (Windows NT 5.0; U) 3, 28 -- X-Accept-Language: fr 3, 28 -- MIME-Version: 1.0 3, 28 -- To: Microscopy-at-microscopy.com 3, 28 -- Subject: looking for 3D reconstruction software 3, 28 -- Content-Type: text/plain; charset=us-ascii 3, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 18, 24 -- From j.bilde-at-risoe.dk Thu May 18 02:52:51 2006 18, 24 -- Received: from mail.risoe.dk (mail.risoe.dk [130.226.48.21]) 18, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4I7qpYh020811 18, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 02:52:51 -0500 18, 24 -- Received: from EXCHG-VS1.risoe.dk (exchg-01.risoe.dk [10.0.0.26]) 18, 24 -- by risoe.dk (PMDF V6.2-X26 #31094) with ESMTP id {USJ508Q11SQCWB013K-at-risoe.dk} 18, 24 -- for Microscopy-at-microscopy.com; Thu, 18, 24 -- 18 May 2006 09:52:58 +0200 (Romance Daylight Time) 18, 24 -- Date: Thu, 18 May 2006 09:52:06 +0200 18, 24 -- From: j.bilde-at-risoe.dk 18, 24 -- Subject: RE: [Microscopy] looking for 3D reconstruction software 18, 24 -- To: ludovic.pinier-at-thalesgroup.com 18, 24 -- Cc: Microscopy-at-microscopy.com 18, 24 -- Message-id: {6463F256A85DC14CAB07F087A247997232D881-at-EXCHG-VS1.risoe.dk} 18, 24 -- MIME-version: 1.0 18, 24 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 18, 24 -- Content-type: text/plain; charset=iso-8859-1 18, 24 -- Thread-Topic: [Microscopy] looking for 3D reconstruction software 18, 24 -- Thread-Index: AcZ6SoQPEdPyl/6WTPGN/T6uXN/ZHQABKOKg 18, 24 -- Content-class: urn:content-classes:message 18, 24 -- X-MS-Has-Attach: 18, 24 -- X-MS-TNEF-Correlator: 18, 24 -- Content-Transfer-Encoding: 8bit 18, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4I7qpYh020811 ==============================End of - Headers==============================
I'll second the notion that your problem is static. In my lab here in Houston, I have the same problem when there is low humidity. On days of high humidity, no "run around" sections.
Mannie Steglich UTMD Anderson Cancer Center
kempfsc-at-auburn.edu
05/17/2006 07:41 PM Please respond to kempfsc
To: msteglic-at-mdanderson.org cc:
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Email: kempfsc-at-auburn.edu Name: Steve Kempf
Organization: Auburn University
Title-Subject: [Filtered] Running away sections
Question: I've had the strangest thing happen this past weekend and since and no one who I've talked to (some very experienced with thin sectioning) has been able to help.
I had been sectioning a block of tissue embedded in Poly/Bed 812 at 50 nm without difficulty using a Diatome diamond knife on a Leica Ultracut microtome, getting beautiful silver sections, spredding then by wafting a toothpick dipped in chloroform over them, and picking them up on formvar coated slot grids. All has been well until this weekend. Everything was set-up as it had been the previous day. Same knife, same boat water, same toothpicks, same choroform, same block, etc. As was the case previously, I got beautiful silver sections, but when I when to spread them with chloroform on a toothpick, they ran away as I moved the toothpick close to them, lickety-split. I could chase them around the boat with the toothpick. When I tried to pick them up on a grid, I couldn't. They would just slide off, back onto the water surface in the boat. Arrrrgh! Naturally, this is happening just as I reach the critical spot in the tissue I'm sectioning.
Not to be deterred, I cleaned the boat by washing it with clean water, got new beakers to hold the water I use, and tried again. The same thing happened. At that point I called it quits for the day and hoped it was some strange environmental effect that would disappear the following week. No Joy! When I sat down to section today, the same thing happened. I tried cleaning the knife boat with 0.2% ethanol in water, got new glassware, new chloroform, new toothpicks, etc. Still no joy?
So, my question is, can anyone give me some guidance as to what might be going on? I'm at my wits end.
Oh, one other thing, if I let the chloroform evaporate off the dipped toothpick, the sections don't run away from it.
Yesterday was the first time that I tried to cut a "turbinate" nose bone, and I found it extremely hard, and even the poor razor blades were instantly dull when I was trying to trim the block.
Does anyone have any good decalcification schemes for processing of specimens which contain bone for electron microscopy?
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==============================Original Headers============================== 4, 20 -- From GBurgess-at-exchange.hsc.mb.ca Thu May 18 08:17:32 2006 4, 20 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IDHVhQ024807 4, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 08:17:32 -0500 4, 20 -- Received: from hscxntmx0007.hsc.mb.ca (unverified [172.16.6.179]) by 4, 20 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 4, 20 -- {B0020730836-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 4, 20 -- 18 May 2006 08:17:35 -0500 4, 20 -- Received: by hscxntmx0007.hsc.mb.ca with Internet Mail Service 4, 20 -- (5.5.2653.19)id {DXXN75Q9} ; Thu, 18 May 2006 08:18:54 -0500 4, 20 -- Message-ID: 4, 20 -- {00A937989100304A83A058F6C45873FF32A3F8-at-hscxntmx0005.hsc.mb.ca} 4, 20 -- Date: Thu, 18 May 2006 08:12:39 -0500 4, 20 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 4, 20 -- Subject: Decalcification of bone 4, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 4, 20 -- X-Mailer: Internet Mail Service (5.5.2653.19) 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: text/plain; 4, 20 -- charset="iso-8859-1" ==============================End of - Headers==============================
Steve, We gave up smelling chloroform a while ago and switched to using a hot wire. The supply houses sell these, very nice units, but with a little care you could probably make one. They deliver a controlled amount of heat to a thin wire loop. You wave this near the sections and just like chloroform, the heat flattens the sections right out. I realize this isn't a fix for your problem right now, but something to consider for the future. Your liver may thank you!
As ever, Tobias Baskin } } Email: kempfsc-at-auburn.edu } Name: Steve Kempf } } Organization: Auburn University } } Title-Subject: [Filtered] Running away sections } } Question: I've had the strangest thing happen this past weekend and } since and no one who I've talked to (some very experienced with thin } sectioning) has been able to help. } } I had been sectioning a block of tissue embedded in Poly/Bed 812 at } 50 nm without difficulty using a Diatome diamond knife on a Leica } Ultracut microtome, getting beautiful silver sections, spredding } then by wafting a toothpick dipped in chloroform over them, and } picking them up on formvar coated slot grids. All has been well } until this weekend. Everything was set-up as it had been the } previous day. Same knife, same boat water, same toothpicks, same } choroform, same block, etc. As was the case previously, I got } beautiful silver sections, but when I when to spread them with } chloroform on a toothpick, they ran away as I moved the toothpick } close to them, lickety-split. I could chase them around the boat } with the toothpick. When I tried to pick them up on a grid, I } couldn't. They would just slide off, back onto the water surface in } the boat. Arrrrgh! Naturally, this is happening just as I reach the } critical spot in the tissue I'm sectioning. } } Not to be deterred, I cleaned the boat by washing it with clean } water, got new beakers to hold the water I use, and tried again. The } same thing happened. At that point I called it quits for the day and } hoped it was some strange environmental effect that would disappear } the following week. No Joy! When I sat down to section today, the } same thing happened. I tried cleaning the knife boat with 0.2% } ethanol in water, got new glassware, new chloroform, new toothpicks, } etc. Still no joy? } } So, my question is, can anyone give me some guidance as to what } might be going on? I'm at my wits end. } } Oh, one other thing, if I let the chloroform evaporate off the } dipped toothpick, the sections don't run away from it. } } Thanks for any advice, } } Steve } } -
This is a perfect question for Histonet: www.histonet.org to subscribe to the mailing list. There are several "boneheads" on the list who can help you.
Phil
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 4, 23 -- From oshel1pe-at-cmich.edu Thu May 18 08:39:03 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 k4IDd3HH012474 4, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 08:39:03 -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 k4IE8X5X020057; 4, 23 -- Thu, 18 May 2006 10:14:08 -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 -- Thu, 18 May 2006 09:33:27 -0400 4, 23 -- Mime-Version: 1.0 4, 23 -- Message-Id: {f06230904c09227ede4c3-at-[141.209.160.132]} 4, 23 -- In-Reply-To: {200605181321.k4IDLYSe032565-at-ns.microscopy.com} 4, 23 -- References: {200605181321.k4IDLYSe032565-at-ns.microscopy.com} 4, 23 -- Date: Thu, 18 May 2006 09:33:27 -0400 4, 23 -- To: GBurgess-at-exchange.hsc.mb.ca 4, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 23 -- Subject: Re: [Microscopy] Decalcification of bone 4, 23 -- Cc: Microscopy-at-microscopy.com 4, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 23 -- X-OriginalArrivalTime: 18 May 2006 13:33:27.0876 (UTC) FILETIME=[A543D840:01C67A7F] 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==============================
Although Histo folks use formic acid, it is murder on ultrastructure.
I have better luck with small bone by 1st fixing for an hour or two in regular fixative, then soaking in a buffered EDTA solution overnight, then refixing in Glut, then embed as normal or in a hard epoxy.
Still... Don't use your BEST knife
Lou Ann
On May 18, 2006, at 8:18 AM, GBurgess-at-exchange.hsc.mb.ca wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } } Yesterday was the first time that I tried to cut a "turbinate" nose } bone, } and I found it extremely hard, and even the poor razor blades were } instantly } dull when I was trying to trim the block. } } Does anyone have any good decalcification schemes for processing of } specimens which contain bone for electron microscopy? } } This e-mail and/or any documents in this transmission is intended for } the address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying } or dissemination is strictly prohibited. If you receive this } transmission in error, please notify the sender immediately and return } the original. } } ==============================Original } Headers============================== } 4, 20 -- From GBurgess-at-exchange.hsc.mb.ca Thu May 18 08:17:32 2006 } 4, 20 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca } [142.233.100.122]) } 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4IDHVhQ024807 } 4, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 08:17:32 } -0500 } 4, 20 -- Received: from hscxntmx0007.hsc.mb.ca (unverified } [172.16.6.179]) by } 4, 20 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP } id } 4, 20 -- {B0020730836-at-hscxntmx0003.hsc.mb.ca} for } {Microscopy-at-microscopy.com} ;Thu, } 4, 20 -- 18 May 2006 08:17:35 -0500 } 4, 20 -- Received: by hscxntmx0007.hsc.mb.ca with Internet Mail Service } 4, 20 -- (5.5.2653.19)id {DXXN75Q9} ; Thu, 18 May 2006 08:18:54 -0500 } 4, 20 -- Message-ID: } 4, 20 -- } {00A937989100304A83A058F6C45873FF32A3F8-at-hscxntmx0005.hsc.mb.ca} } 4, 20 -- Date: Thu, 18 May 2006 08:12:39 -0500 } 4, 20 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} } 4, 20 -- Subject: Decalcification of bone } 4, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} } 4, 20 -- X-Mailer: Internet Mail Service (5.5.2653.19) } 4, 20 -- MIME-Version: 1.0 } 4, 20 -- Content-Type: text/plain; } 4, 20 -- charset="iso-8859-1" } ==============================End of - } Headers============================== } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Lou Ann Miller, MT(ASCP) Service Supervisor Center for Microscopic Imaging College of Veterinary Medicine Rm 1204 VMBSB 2001 S Lincoln Ave Urbana, IL 61802
217/244-1567
==============================Original Headers============================== 9, 19 -- From lamiller-at-uiuc.edu Thu May 18 09:27:24 2006 9, 19 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu [128.174.5.96]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IEROn5023775 9, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:27:24 -0500 9, 19 -- Received: from [130.126.19.3] (tortoise.cvm.uiuc.edu [130.126.19.3]) 9, 19 -- by expredir5.cites.uiuc.edu (8.13.6/8.13.6) with ESMTP id k4IERJJY019090; 9, 19 -- Thu, 18 May 2006 09:27:19 -0500 (CDT) 9, 19 -- In-Reply-To: {200605181318.k4IDIAhw025985-at-ns.microscopy.com} 9, 19 -- References: {200605181318.k4IDIAhw025985-at-ns.microscopy.com} 9, 19 -- Mime-Version: 1.0 (Apple Message framework v624) 9, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 19 -- Message-Id: {5877281a06fcac3df10353e871658681-at-uiuc.edu} 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- Cc: Microscopy-at-microscopy.com 9, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} 9, 19 -- Subject: Re: [Microscopy] Decalcification of bone 9, 19 -- Date: Thu, 18 May 2006 09:27:19 -0500 9, 19 -- To: GBurgess-at-exchange.hsc.mb.ca 9, 19 -- X-Mailer: Apple Mail (2.624) ==============================End of - Headers==============================
I'm seeking advice from those who have had success corralling small specimens into dialysis tubing for EM processing. We recently tried some samples in 2 different types of tubing that were convenient just because we had them around. The samples were high pressure frozen and freeze substituted but in the end one type of tube just dissolved in the acetone and the other shrunk greatly, crushing the contents. Does anyone have a recommendation for a brand of tubing that is robust in this kind of treatment?
Our thought has been to embed the entire tube and contents for sectioning. Is there any reason that might not work?
Thanks in advance, Jay
Jay Campbell Research Specialist University of Wisconsin Laboratory of Molecular Biology R.M. Bock Labs 1525 Linden Drive Madison, WI 53706 jmcampbe-at-wisc.edu 608 263 8481
==============================Original Headers============================== 7, 23 -- From microtomy-at-gmail.com Thu May 18 09:38:51 2006 7, 23 -- Received: from py-out-1112.google.com (py-out-1112.google.com [64.233.166.178]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IEcoPK001433 7, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:38:51 -0500 7, 23 -- Received: by py-out-1112.google.com with SMTP id 39so594352pyu 7, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 07:38:49 -0700 (PDT) 7, 23 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 23 -- s=beta; d=gmail.com; 7, 23 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 7, 23 -- b=qNXPebIo1zwv4ZNEkldjDyh372J3FFcw3983dnRLfSL4tuvxzmZMUGUCx1SI8YpBuiaaQN/7XC5kWf2t1LrJgeoD8UZj024dF+YeGykKLUUuLegHJqvLVIIdsua3CLJpYGS6TVZrI1yMuC3xsEctloWrMVjlUx7xebDjzj8IRUY= 7, 23 -- Received: by 10.35.127.7 with SMTP id e7mr631604pyn; 7, 23 -- Thu, 18 May 2006 07:38:49 -0700 (PDT) 7, 23 -- Received: by 10.35.93.2 with HTTP; Thu, 18 May 2006 07:38:48 -0700 (PDT) 7, 23 -- Message-ID: {ef6bda5c0605180738h2d95c6depae41cd14f56523fe-at-mail.gmail.com} 7, 23 -- Date: Thu, 18 May 2006 09:38:48 -0500 7, 23 -- From: "Jay Campbell" {microtomy-at-gmail.com} 7, 23 -- To: Microscopy-at-microscopy.com 7, 23 -- Subject: Dialysis tubing for processing 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 23 -- Content-Disposition: inline 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4IEcoPK001433 ==============================End of - Headers==============================
Following 3 days of incubation with a substance, I can observe an important decrease in cell viability by ELISA (resazurin assay, BTW I recommand this test since it is very good and costs nothing). Now I would like to know if the cells die by apopotosis. Unfortunately the commercial kits are very expensive and there seems to be no "home made" solution. I thought about observing the cells in fluorescence using DAPI. I know DNA compaction is a late event of apoptosis, and that microcroscopy does not give quantitative measurements (easily) but it could at least show a clear effect, like Y/N apoptosis is involved in the reduction of cell viability. If I see no sign of apoptosis by fluorescence I can search in another direction and save some money in the process.
I would like to read your comments about my idea.
Regards,
Stéphane (without-the-i)
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==============================Original Headers============================== 8, 18 -- From nizets2-at-yahoo.com Thu May 18 09:53:17 2006 8, 18 -- Received: from web37404.mail.mud.yahoo.com (web37404.mail.mud.yahoo.com [209.191.87.57]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4IErFJt011500 8, 18 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 09:53:16 -0500 8, 18 -- Received: (qmail 96776 invoked by uid 60001); 18 May 2006 14:53:13 -0000 8, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 18 -- s=s1024; d=yahoo.com; 8, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 18 -- b=U9UbHsrECgn8UeEPj1p3NVEWyPNYkks74nv/xPtx7g9omymUD6lbxNdiwgOQJn/OL0I2iMVb7YNCfccr1aX0X2e32YSd+SNjKs6/Ig13OP9PuQ+mrsBpHKC3xBymgIQlXwgRKSY9xQ0ruxUClxR4XwxphUrtXh9MtRvjvzPPxx4= ; 8, 18 -- Message-ID: {20060518145313.96774.qmail-at-web37404.mail.mud.yahoo.com} 8, 18 -- Received: from [80.122.101.102] by web37404.mail.mud.yahoo.com via HTTP; Thu, 18 May 2006 07:53:13 PDT 8, 18 -- Date: Thu, 18 May 2006 07:53:13 -0700 (PDT) 8, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 18 -- Subject: apoptosis, DAPI and fluorescence microscopy 8, 18 -- To: microscopy-at-microscopy.com 8, 18 -- MIME-Version: 1.0 8, 18 -- Content-Type: text/plain; charset=iso-8859-1 8, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
We use a Diatome Static LineII (which is an anti static device) with the tip hanging close to where the sections are cut. I use it for room temperature slicing even though it was designed for Cryo slicing. We bought ours through EMS at www.emsdiasum.com. Hope this is helpful. shawn
==============================Original Headers============================== 2, 27 -- From trent-at-ornl.gov Thu May 18 11:39:24 2006 2, 27 -- Received: from emroute1.ornl.gov (emroute1.ornl.gov [160.91.4.119]) 2, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IGdNY2024207 2, 27 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 11:39:24 -0500 2, 27 -- Received: from emroute1.ornl.gov (localhost [127.0.0.1]) 2, 27 -- by emroute1.ornl.gov (PMDF V6.2-1x9 #31038) 2, 27 -- with ESMTP id {0IZG0038GZLLX8-at-emroute1.ornl.gov} for 2, 27 -- microscopy-at-microscopy.com; Thu, 18 May 2006 12:39:21 -0400 (EDT) 2, 27 -- Received: from ORNLEXCHANGE.ornl.gov (ornlexchange1.ornl.gov [160.91.1.20]) 2, 27 -- by emroute1.ornl.gov (PMDF V6.2-1x9 #31038) 2, 27 -- with ESMTP id {0IZG00564ZLLIJ-at-emroute1.ornl.gov} for 2, 27 -- microscopy-at-microscopy.com; Thu, 18 May 2006 12:39:21 -0400 (EDT) 2, 27 -- Received: from 160.91.157.36 ([160.91.157.36]) 2, 27 -- by ORNLEXCHANGE.ornl.gov ([160.91.1.32]) via Exchange Front-End Server 2, 27 -- mail.ornl.gov ([160.91.4.25]) with Microsoft Exchange Server HTTP-DAV ; Thu, 2, 27 -- 18 May 2006 16:39:20 +0000 2, 27 -- Date: Thu, 18 May 2006 12:39:20 -0400 2, 27 -- From: Shawn Reeves {trent-at-ornl.gov} 2, 27 -- Subject: Re: viaWWW: Running away sections 2, 27 -- To: microscopy-at-microscopy.com 2, 27 -- Message-id: {C0921B78.86E%trent-at-ornl.gov} 2, 27 -- MIME-version: 1.0 2, 27 -- Content-type: text/plain; charset=US-ASCII 2, 27 -- Content-transfer-encoding: 7bit 2, 27 -- User-Agent: Microsoft-Entourage/11.2.3.060209 2, 27 -- Thread-Topic: viaWWW: Running away sections 2, 27 -- Thread-Index: AcZ6mZxy2xQI2OaMEdqD4gAKlebEPA== ==============================End of - Headers==============================
On May 17, 2006, at 5:38 PM, Mayya_Saab-at-dps-consulting.com wrote:
} Question: What is this field? How can it help an individual? } Dear Mayya, Dark field imaging is a technique where the unscattered beam (of either light or electrons) is removed from the image, so that the image consists only of the scattered particles. With this technique the most strongly scattering parts of the specimen appear brightest. It is a help for specimens of low contrast, and it can be used to highlight those parts of a specimen that scatter into well-defined areas; e.g., crystals having a particular orientation or heavy atoms. 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 May 18 11:59:53 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 k4IGxqhx002064 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 18 May 2006 11:59:53 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP 4, 22 -- id 18FC73650A; Thu, 18 May 2006 09:59:51 -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 93F21349E8; Thu, 18 May 2006 09:59:47 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200605180038.k4I0c55P018887-at-ns.microscopy.com} 4, 22 -- References: {200605180038.k4I0c55P018887-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {fd687a5a9069f6113fbd898f56217b75-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] AskAMicroscopist: what is dark field ? 4, 22 -- Date: Thu, 18 May 2006 10:10:37 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com, Mayya_Saab-at-dps-consulting.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Here in Britain we have a gov't TV advertisement aimed at recruiting teachers, showing young people with "active minds" asking lots of awkward questions. One is:
"What's the difference between dust and fluff?"
WE know the answer - there should be one in every classroom!
*** from the neutrino who always pulls his weight ***,
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
==============================Original Headers============================== 7, 21 -- From hinmeigeng-at-hotmail.com Thu May 18 12:02:40 2006 7, 21 -- Received: from hotmail.com (bay101-f8.bay101.hotmail.com [64.4.56.18]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IH2eM3006754 7, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 12:02:40 -0500 7, 21 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 7, 21 -- Thu, 18 May 2006 10:02:39 -0700 7, 21 -- Message-ID: {BAY101-F865EAC18D58EF4D9E5967CAA60-at-phx.gbl} 7, 21 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 7, 21 -- Thu, 18 May 2006 17:02:36 GMT 7, 21 -- X-Originating-IP: [86.128.212.236] 7, 21 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 7, 21 -- X-Sender: hinmeigeng-at-hotmail.com 7, 21 -- Reply-To: R.H.Olley-at-reading.ac.uk 7, 21 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 7, 21 -- To: Microscopy-at-MSA.Microscopy.Com 7, 21 -- Cc: R.H.Olley-at-reading.ac.uk 7, 21 -- Subject: Dust & Fluff 7, 21 -- Date: Thu, 18 May 2006 17:02:36 +0000 7, 21 -- Mime-Version: 1.0 7, 21 -- Content-Type: text/plain; format=flowed 7, 21 -- X-OriginalArrivalTime: 18 May 2006 17:02:39.0473 (UTC) FILETIME=[DE998E10:01C67A9C] ==============================End of - Headers==============================
} -------------- } } For a free program for 3D surface topography have a look at } } Measuring Surface Topography with Scanning Electron } Microscopy. I. EZEImage: A Program to Obtain 3D Surface Data } Ezequiel Ponz,1 Juan Luis Ladaga,2 and Rita Dominga Bonetto1* } } which appeared in Microscopy and Microanalysis 12 (2006) 170-177. } } Best regards, Jřrgen. } } = = = = = = = = = = = = = = = = = } } Joergen B. Bilde-Soerensen } Senior Research Scientist, Ph. D. } Materials Research Department } Risoe National Laboratory } DK-4000 Roskilde } Denmark } } e-mail: j.bilde-at-risoe.dk } phone: +45 4677 5802 (direct) } phone: +45 4677 4677 (switchboard) } fax: +45 4677 5758 } website: http://www.risoe.dk/afm/Personal/jqbi/jqbi.htm
Did anybody tried to use this program? I would greatly appreciate any comments. Examples from the paper were not convincing. I still do not believe that gray level alone could be used for successful 3D reconstruction. I would be glad to change my opinion.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 371 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
10% EDTA adjusted with NaOH to PH 7.5 will do the work. Change solution every other day for 1-2 weeks (for small pieces of bone, about 0.5 mm square). Better on a rotator.
As for calcified bone, I trim it with a small file in a fume hood.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 371 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -------------- } } } Yesterday was the first time that I tried to cut a } "turbinate" nose bone, and I found it extremely hard, and } even the poor razor blades were instantly dull when I was } trying to trim the block. } } Does anyone have any good decalcification schemes for } processing of specimens which contain bone for electron microscopy? } } This e-mail and/or any documents in this transmission is } intended for the address(s) only and may contain legally } privileged or confidential information. Any unauthorized use, } disclosure, distribution, copying or dissemination is } strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original.
==============================Original Headers============================== 6, 23 -- From DusevichV-at-umkc.edu Thu May 18 13:37:19 2006 6, 23 -- Received: from kc-msxproto3.kc.umkc.edu (kc-msxproto3.kc.umkc.edu [134.193.44.10]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IIbJj9001164 6, 23 -- for {microscopy-at-msa.microscopy.com} ; Thu, 18 May 2006 13:37:19 -0500 6, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by kc-msxproto3.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Thu, 18 May 2006 13:37:19 -0500 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: RE: [Microscopy] Decalcification of bone 6, 23 -- Date: Thu, 18 May 2006 13:37:18 -0500 6, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADC1B-at-KC-MSX1.kc.umkc.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [Microscopy] Decalcification of bone 6, 23 -- Thread-Index: AcZ6fYQBSS+gU14uRM6GvVwpFipGdwAKfJaQ 6, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 6, 23 -- To: {GBurgess-at-exchange.hsc.mb.ca} , {microscopy-at-msa.microscopy.com} 6, 23 -- X-OriginalArrivalTime: 18 May 2006 18:37:19.0425 (UTC) FILETIME=[181D7310:01C67AAA] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4IIbJj9001164 ==============================End of - Headers==============================
Distilled water does have a slightly acidic pH. While we use distilled water in our cooling systems, we add a bit of sodium bicarbonate to bring the pH up to neutral. I just pull out the pH paper 3-4 times a year and check the cooling systems.
Cheers, Henk
At 08:02 PM 05/17/06, tivol-at-caltech.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
==============================Original Headers============================== 9, 24 -- From colijn.1-at-osu.edu Thu May 18 13:58:31 2006 9, 24 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IIwVWW011409 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 18 May 2006 13:58:31 -0500 9, 24 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 9, 24 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 9, 24 -- id {01M2KKZE73QO9V7GMH-at-er6s1.eng.ohio-state.edu} for 9, 24 -- Microscopy-at-microscopy.com; Thu, 18 May 2006 14:58:29 -0400 (EDT) 9, 24 -- Received: from HOC1.ecr6.ohio-state.edu 9, 24 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 9, 24 -- (PMDF V6.2-1x11 #31056) 9, 24 -- with ESMTPA id {01M2KKZDCVWA9VQZHV-at-er6s1.eng.ohio-state.edu} for 9, 24 -- Microscopy-at-microscopy.com; Thu, 18 May 2006 14:58:28 -0400 (EDT) 9, 24 -- Date: Thu, 18 May 2006 14:59:03 -0400 9, 24 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 9, 24 -- Subject: Re: [Microscopy] thanks for the info - TEM water recirculator 9, 24 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 9, 24 -- To: Microscopy-at-microscopy.com 9, 24 -- Message-id: {7.0.1.0.2.20060518145847.04b58500-at-osu.edu} 9, 24 -- Message-id: {7.0.1.0.2.20060517204237.03e92c98-at-osu.edu} 9, 24 -- MIME-version: 1.0 9, 24 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 9, 24 -- X-Env-From: auth/colijn.1-at-osu.edu ==============================End of - Headers==============================
We are doing a rush job for a client who requires 4.0 um sections from JB 4 blocks. Our ultramicrotomists extraordinaire, Cheryl, is having a dickens of a time getting the sections to remain flat when removing them from the knife. She is cutting on glass and taking sections from the dry edge with a fine forceps. As soon as the sections leave the knife, they curl and won't uncurl when placed on a drop of water on a slide.
Not only is this a rush job in support of a grant proposal, but it requires serial sectioning with no missing sections, and we have, like, no real experience with this resin. Cheryl has tried various sized block faces and different thicknesses for the sections, but nothing is helping.
The thumping sound you hear is a head hitting a wall----repeatedly. Can anyone HEEEELLLLP??
Thanks!
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============================== 11, 23 -- From TindallR-at-missouri.edu Thu May 18 14:15:28 2006 11, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 11, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IJFS1G021560 11, 23 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 14:15:28 -0500 11, 23 -- Received: from UM-EMAIL10.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 11, 23 -- Thu, 18 May 2006 14:15:27 -0500 11, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 11, 23 -- Content-class: urn:content-classes:message 11, 23 -- MIME-Version: 1.0 11, 23 -- Content-Type: text/plain; 11, 23 -- charset="us-ascii" 11, 23 -- Subject: Sectioning JB 4 resin 11, 23 -- Date: Thu, 18 May 2006 14:15:26 -0500 11, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE701849FD1-at-UM-EMAIL09.um.umsystem.edu} 11, 23 -- X-MS-Has-Attach: 11, 23 -- X-MS-TNEF-Correlator: 11, 23 -- Thread-Topic: Sectioning JB 4 resin 11, 23 -- Thread-Index: AcZ6r2r/IkxeShTKSq2TzE/9tevkIQ== 11, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 11, 23 -- To: {microscopy-at-microscopy.com} 11, 23 -- X-OriginalArrivalTime: 18 May 2006 19:15:27.0809 (UTC) FILETIME=[6C192310:01C67AAF] 11, 23 -- Content-Transfer-Encoding: 8bit 11, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4IJFS1G021560 ==============================End of - Headers==============================
The problem of dealing with algal growth in water chillers is discussed in some detail on p. 216 of my book, Vacuum Methods in Electron Microscopy (ISBN 1-85578-052-6, available from SPi, Ladd, Pella, etc.). One basic fact to remember is that algae require light to grow, and so you can go a long way toward reducing algal growth by excluding light from the chiller system (a light-tight cover for the reservoir and opaque tubing leading to and from the instrument). Beyond that, it is simple to add a bit of an algacide such as Chloroamine-T or dichlorophene to the system. These chemicals are available from specialty chemical companies such as Polysciences, Sigma, etc., and perhaps from your local air conditioning or swimming pool service company. For Chloramine-T the recommended concentration is about one gram per gallon of water. It is, of course, advisable to use distilled water to avoid the build-up of scale in the system. -- 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============================== 1, 14 -- From bigelow-at-engin.umich.edu Thu May 18 14:22:18 2006 1, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IJMIh3031472 1, 14 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 14:22:18 -0500 1, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- by srvr22.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k4IJMHsH012165 1, 14 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 15:22:17 -0400 (EDT) 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06210201c09276702cb1-at-[141.212.131.221]} 1, 14 -- Date: Thu, 18 May 2006 15:22:16 -0400 1, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 1, 14 -- Subject: [Microscopy] RE: Algae in water chillers 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
ah yes, the joys of bmma. i have spent many a pleasant hour cursing the curling sections. in my experience, no change in shape, speed, thickness makes a difference. I learned to be waiting for the section to start cutting and then either grabbing a corner of it with a fine forceps or using the forceps' tines to hold the corner down on to the surface of the knife while it cut. then stopping the microtome, removing the section, and re-starting the cutting motor. very tedious and time-consuming. it does help to listen to NPR while doing this. I know you are stuck to the whims of your client whose blocks are already embedded but I strongly recommend any fans of JB-4 consider switching to the generic (and therefore less expensive) butylmethyl methacrylate resin mix of Tobias Baskin. you can cut on water filled boats and use acetone to extract the resin so the sensitivity is better and the sectioning is trivial. My condolences to Cheryl. Tom
At 02:16 PM 05/18/06, 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
Whoops - I mistakenly typed the wrong resin name in the first sentence of my last posting - the correct version is below:
} ah yes, the joys of JB-4. i have spent many a pleasant hour cursing the } curling sections. in my experience, no change in shape, speed, thickness } makes a difference. I learned to be waiting for the section to start } cutting and then either grabbing a corner of it with a fine forceps or } using the forceps' tines to hold the corner down on to the surface of the } knife while it cut. then stopping the microtome, removing the section, and } re-starting the cutting motor. very tedious and time-consuming. it does } help to listen to NPR while doing this. I know you are stuck to the whims } of your client whose blocks are already embedded but I strongly recommend } any fans of JB-4 consider switching to the generic (and therefore less } expensive) butylmethyl methacrylate resin mix of Tobias Baskin. you can } cut on water filled boats and use acetone to extract the resin so the } sensitivity is better and the sectioning is trivial. My condolences to } Cheryl. Tom } } At 02:16 PM 05/18/06, you wrote: } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Dear Listers, } } } } We are doing a rush job for a client who requires 4.0 um sections from } } JB 4 blocks. Our ultramicrotomists extraordinaire, Cheryl, is having a } } dickens of a time getting the sections to remain flat when removing them } } from the knife. She is cutting on glass and taking sections from the } } dry edge with a fine forceps. As soon as the sections leave the knife, } } they curl and won't uncurl when placed on a drop of water on a slide. } } } } Not only is this a rush job in support of a grant proposal, but it } } requires serial sectioning with no missing sections, and we have, like, } } no real experience with this resin. Cheryl has tried various sized } } block faces and different thicknesses for the sections, but nothing is } } helping. } } } } The thumping sound you hear is a head hitting a wall----repeatedly. Can } } anyone HEEEELLLLP?? } } } } Thanks! } } } } 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============================== } } 11, 23 -- From TindallR-at-missouri.edu Thu May 18 14:15:28 2006 } } 11, 23 -- Received: from um-exproto9.um.umsystem.edu } } (um-exproto9.um.umsystem.edu [207.160.151.49]) } } 11, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } } id k4IJFS1G021560 } } 11, 23 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 14:15:28 } } -0500 } } 11, 23 -- Received: from UM-EMAIL10.um.umsystem.edu ([207.160.151.31]) by } } um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } } 11, 23 -- Thu, 18 May 2006 14:15:27 -0500 } } 11, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } } 11, 23 -- Content-class: urn:content-classes:message } } 11, 23 -- MIME-Version: 1.0 } } 11, 23 -- Content-Type: text/plain; } } 11, 23 -- charset="us-ascii" } } 11, 23 -- Subject: Sectioning JB 4 resin } } 11, 23 -- Date: Thu, 18 May 2006 14:15:26 -0500 } } 11, 23 -- Message-ID: } } {BA876152E8653240BE8572E897083EE701849FD1-at-UM-EMAIL09.um.umsystem.edu} } } 11, 23 -- X-MS-Has-Attach: } } 11, 23 -- X-MS-TNEF-Correlator: } } 11, 23 -- Thread-Topic: Sectioning JB 4 resin } } 11, 23 -- Thread-Index: AcZ6r2r/IkxeShTKSq2TzE/9tevkIQ== } } 11, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } } 11, 23 -- To: {microscopy-at-microscopy.com} } } 11, 23 -- X-OriginalArrivalTime: 18 May 2006 19:15:27.0809 (UTC) } } FILETIME=[6C192310:01C67AAF] } } 11, 23 -- Content-Transfer-Encoding: 8bit } } 11, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id k4IJFS1G021560 } } ==============================End of - Headers============================== } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 2 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } } } ==============================Original Headers============================== } 9, 20 -- From PhillipsT-at-missouri.edu Thu May 18 14:29:25 2006 } 9, 20 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k4IJTPX2008984 } 9, 20 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 18 May 2006 } 14:29:25 -0500 } 9, 20 -- Received: from um-exproto9.um.umsystem.edu ([207.160.151.148]) by } um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 9, 20 -- Thu, 18 May 2006 14:29:25 -0500 } 9, 20 -- Received: from phillips-dell.missouri.edu ([128.206.81.132]) by } um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft } SMTPSVC(6.0.3790.1830); } 9, 20 -- Thu, 18 May 2006 14:29:24 -0500 } 9, 20 -- Message-Id: {6.0.0.22.2.20060518142258.047861a0-at-pop.missouri.edu} } 9, 20 -- X-Sender: phillipst-at-pop.missouri.edu } 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.0.0.22 } 9, 20 -- Date: Thu, 18 May 2006 14:29:22 -0500 } 9, 20 -- To: Microscopy-at-msa.microscopy.com } 9, 20 -- From: Tom Phillips {phillipst-at-missouri.edu} } 9, 20 -- Subject: Re: [Microscopy] Sectioning JB 4 resin } 9, 20 -- In-Reply-To: {200605181916.k4IJG7it023109-at-ns.microscopy.com} } 9, 20 -- References: {200605181916.k4IJG7it023109-at-ns.microscopy.com} } 9, 20 -- Mime-Version: 1.0 } 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } 9, 20 -- X-OriginalArrivalTime: 18 May 2006 19:29:24.0580 (UTC) } FILETIME=[5EDA4240:01C67AB1] } ==============================End of - Headers==============================
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
} } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Postdoctoral Position in Scanning Transmission Electron Microscopy - } University of California, Santa Barbara } } Applicants should have demonstrated experience in TEM and a strong } background in materials science and diffraction. Preference will be } given to applicants with expertise in STEM techniques, such as } atomic resolution HAADF imaging. Facilities at UCSB include a } Tecnai F30U TEM/STEM and other state-of-the-art imaging, } spectroscopy and diffraction facilities. Research projects include } the characterization of interfaces and defects in high-permittivity } oxide thin films, including gate dielectrics and ferroelectrics. } } The position is available summer/fall 2006. Duration about 1-2 } years, salary is commensurate with qualifications. Candidates must } have a Ph.D. in Materials Science or Physics. Interested candidates } should send a curriculum vitae, publication list and the names of } three references with their contact information to: } } Prof. Susanne Stemmer } Materials Department } University of California } Santa Barbara, CA 93106-5050 } Email: stemmer-at-mrl.ucsb.edu } http://www.mrl.ucsb.edu/~stemmer } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 3, 23 -- From xin-at-magnet.fsu.edu Thu May 18 15:08:08 2006 3, 23 -- Received: from mail.magnet.fsu.edu (mail.magnet.fsu.edu [146.201.250.9]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IK88q1029486 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 15:08:08 -0500 3, 23 -- Received: from xinlt.magnet.fsu.edu (a2-dhcp073.magnet.fsu.edu [146.201.233.73]) 3, 23 -- (authenticated bits=0) 3, 23 -- by mail.magnet.fsu.edu (8.13.6/8.13.6) with ESMTP id k4IK83eR000322 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 18 May 2006 16:08:03 -0400 3, 23 -- Message-Id: {7.0.0.16.2.20060518160544.02267a20-at-magnet.fsu.edu} 3, 23 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.0.16 3, 23 -- Date: Thu, 18 May 2006 16:06:40 -0400 3, 23 -- To: microscopy-at-microscopy.com 3, 23 -- From: Yan Xin {xin-at-magnet.fsu.edu} 3, 23 -- Subject: Postdoctoral Position in Scanning Transmission Electron 3, 23 -- Microscopy 3, 23 -- Mime-Version: 1.0 3, 23 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 3, 23 -- X-NHMFL-MailScanner: Found to be clean 3, 23 -- X-MailScanner-MCPCheck: 3, 23 -- X-NHMFL-MailScanner-SpamCheck: not spam, SpamAssassin (score=-4.905, 3, 23 -- required 4, autolearn=not spam, ALL_TRUSTED -1.80, AWL -0.51, 3, 23 -- BAYES_00 -2.60) 3, 23 -- X-MailScanner-From: xin-at-magnet.fsu.edu ==============================End of - Headers==============================
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Question: I just wanted to thank everyone who replied to my question yesterday. I too thought static charge might be the problem; however, it seemed strange that if that was the case, the sections didn't "run away" when I put the chloroform dipped toothpick next to them after allowing the chloroform to evaporate. At any rate, late yesterday I dismounted the block I had been having the problems with and mounted new one and, guess what, no problems. Everything worked fine. I didn't have time to re-try the block that I had been having the problem with again followng that, so I put it in a 60 degree oven overnight and mounted it this morning. And, as the Gods of sectioning would have it, everything was back to normal with no "running away" problems.
Go figure?
So, if it was static, did just changing the block releave the charge?
At any rate, thanks again for all the suggestions.
I have a client who would like to figure out the composition (in terms of protein, carbohydrates, and nucleic acids) of small (0.2um - 1um) abiotic particles. They have already tried to stain with DAPI and a couple of other things (I don't know what) and look with fluorescence and flow cytometry, but the size/signal is too low.
The only approach I know is microscopy (I'm a one-trick pony). Should they be pursuing other methods, talking to biochemists, geochemists, food nutritionists? Their sample size is very tiny; microscopic.
For microscopy all I can think of is things like antibodies to nucleic acids, hitting it with all kinds of lectins, and other shotgun approaches. They have been able to get some of these on a coated grid with an airfuge; they are pretty electron dense. I shudder to think that I will need to embed and section them, but I can give it a shot. No idea if they also contain metallic particles, but I would not be surprised.
Does anyone have any ideas how to approach this challenge?
Mahalo (thanks), 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============================== 8, 19 -- From tina-at-pbrc.hawaii.edu Thu May 18 17:07:26 2006 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IM7Qns021148 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 17:07:26 -0500 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k4IM7Mf6010712 8, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 12:07:22 -1000 (HST) 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k4IM7Lu2010709 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 12:07:21 -1000 (HST) 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 8, 19 -- Date: Thu, 18 May 2006 12:07:20 -1000 (HST) 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 8, 19 -- X-Sender: tina-at-halia 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 8, 19 -- Subject: Composition of abiotic particles 8, 19 -- Message-ID: {Pine.GSO.4.21.0605181159010.10514-100000-at-halia} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Similar to what Tom suggested below, that is, mechanical restriction to reduce the roll up of the section, is to use a fine tipped artists brush to sort of tamp the section lightly to the surface of the knife as it comes of the knife edge, or to use light pressure on top of section to restrict the roll up. A slow cutting speed would give you more time to do that. Then use the brush to lift it off the knife surface to the slide. I had a microtomist here a year ago who had to do that and I think it was JB-4 or GMA resin that she was cutting. Best of luck to Cheryl!
Gib -- Gilbert (Gib) Ahlstrand, Electron Microscopist, Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
phillipst-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 } ---------------------------------------------------------------------------- } } ah yes, the joys of bmma [JB-4]. i have spent many a pleasant hour cursing the } curling sections. in my experience, no change in shape, speed, thickness } makes a difference. I learned to be waiting for the section to start } cutting and then either grabbing a corner of it with a fine forceps or } using the forceps' tines to hold the corner down on to the surface of the } knife while it cut. then stopping the microtome, removing the section, and } re-starting the cutting motor. very tedious and time-consuming. it does } help to listen to NPR while doing this. I know you are stuck to the whims } of your client whose blocks are already embedded but I strongly recommend } any fans of JB-4 consider switching to the generic (and therefore less } expensive) butylmethyl methacrylate resin mix of Tobias Baskin. you can } cut on water filled boats and use acetone to extract the resin so the } sensitivity is better and the sectioning is trivial. My condolences to } Cheryl. Tom } } At 02:16 PM 05/18/06, you wrote:
} } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Dear Listers, } } } } We are doing a rush job for a client who requires 4.0 um sections from } } JB 4 blocks. Our ultramicrotomists extraordinaire, Cheryl, is having a } } dickens of a time getting the sections to remain flat when removing them } } from the knife. She is cutting on glass and taking sections from the } } dry edge with a fine forceps. As soon as the sections leave the knife, } } they curl and won't uncurl when placed on a drop of water on a slide. } } } } Not only is this a rush job in support of a grant proposal, but it } } requires serial sectioning with no missing sections, and we have, like, } } no real experience with this resin. Cheryl has tried various sized } } block faces and different thicknesses for the sections, but nothing is } } helping. } } } } The thumping sound you hear is a head hitting a wall----repeatedly. Can } } anyone HEEEELLLLP?? } } } } Thanks! } } } } 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 } } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 2 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
==============================Original Headers============================== 5, 19 -- From ahlst007-at-umn.edu Thu May 18 17:53:24 2006 5, 19 -- Received: from mtaout-a.tc.umn.edu (mtaout-a.tc.umn.edu [134.84.119.206]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4IMrOe5031854 5, 19 -- for {Microscopy-at-Microscopy.com} ; Thu, 18 May 2006 17:53:24 -0500 5, 19 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) by mtaout-a.tc.umn.edu with ESMTP for Microscopy-at-Microscopy.com; Thu, 18 May 2006 17:53:23 -0500 (CDT) 5, 19 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 5, 19 -- Message-ID: {446CFAAB.4090105-at-umn.edu} 5, 19 -- Date: Thu, 18 May 2006 17:52:27 -0500 5, 19 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 5, 19 -- Reply-To: ahlst007-at-umn.edu 5, 19 -- Organization: Imaging Center UM 5, 19 -- User-Agent: Thunderbird 1.5.0.2 (Macintosh/20060308) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: Microscopy-at-Microscopy.com 5, 19 -- Subject: Re: [Microscopy] Re: Sectioning JB 4 resin 5, 19 -- References: {200605181931.k4IJVuKX015180-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200605181931.k4IJVuKX015180-at-ns.microscopy.com} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
_____________________________________________ X-from: Randi Olsen Sent: 18. mai 2006 16:04 To: 'Microscopy-at-microscopy.com'
Hi listers,
Our lab wishes to print out b/w images from scanned TEM negatives and bypass completely darkroom enlargements. Printouts should not be expensive but should have a good dynamic range to allow researchers to evaluate fine structures etc. Can anyone suggest (or even recommend) the most suitable monochrome printers for this purpose. We have already a Fujix pictrography system so final (expensive) printouts are no problem here. TIA,
Jim
==============================Original Headers============================== 6, 22 -- From jchalcro-at-neuro.mpg.de Fri May 19 03:21:13 2006 6, 22 -- Received: from neuro.mpg.de (mail.neuro.mpg.de [130.183.250.1]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4J8LCQL029904 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 19 May 2006 03:21:13 -0500 6, 22 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 22 -- Content-class: urn:content-classes:message 6, 22 -- Return-Receipt-To: "James Chalcroft" {jchalcro-at-neuro.mpg.de} 6, 22 -- MIME-Version: 1.0 6, 22 -- Content-Type: text/plain; 6, 22 -- charset="US-ASCII" 6, 22 -- Disposition-Notification-To: "James Chalcroft" {jchalcro-at-neuro.mpg.de} 6, 22 -- Subject: FW: Good grey-scale printer sought for scanned TEM negatives 6, 22 -- Date: Fri, 19 May 2006 10:21:11 +0200 6, 22 -- Message-ID: {6BBC089D4E1E87459875FB80D8030031BE223C-at-s6.neuro.mpg.de} 6, 22 -- X-MS-Has-Attach: 6, 22 -- X-MS-TNEF-Correlator: 6, 22 -- Thread-Topic: Good grey-scale printer sought for scanned TEM negatives 6, 22 -- Thread-Index: AcZ7HPcQ+VpCHhUBQ/GdVvimMZv/3gAABfmw 6, 22 -- From: "James Chalcroft" {jchalcro-at-neuro.mpg.de} 6, 22 -- To: {microscopy-at-msa.microscopy.com} 6, 22 -- Content-Transfer-Encoding: 8bit 6, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4J8LCQL029904 ==============================End of - Headers==============================
I must admit that I have never attempted growing bacteria on grids. But I wonder if the copper of the grids is too reactive. It might both inhibit bacteria and encourage precipitation/reaction products.
You could try a couple of gold grids as a control to see if matters improve.
Good luck
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: Randi.Olsen-at-fagmed.uit.no
Dear Randi,
A possible way out of your problem might be the technique I used years ago in the study of thermophilic bacteria in nature. I deposited carbon films in vacuo on small freshly-cleaved mica squares and let them mature for a few days. The films were carefully floated off on the water surface and the bacteria were allowed to settle on and attach to the underside of the carbon. The films were then transferred to other solutions with a fine Pt loop (or even the original mica piece - submerged then lifted up below the film). Staining with aqueous solutions of UA, AM or PTA can be done either before (preferably) or after the films are picked up with TEM grids. Good luck.
Jim
-----Original Message----- X-from: Randi.Olsen-at-fagmed.uit.no [mailto:Randi.Olsen-at-fagmed.uit.no] Sent: Friday, May 19, 2006 9:49 AM To: James Chalcroft
_____________________________________________ X-from: Randi Olsen Sent: 18. mai 2006 16:04 To: 'Microscopy-at-microscopy.com'
Tina,
If they just need things like "protein" or "carbohydrate", how about cytochemical staining methods for light microscopy? Deposit the particles on slides, and pretend they're bacteria. SEM/EDX ought to pick up metals. Mind, there are things like ninhydrin for protein, and the protein/carbohydrate/fat methods physiological ecologists (and in the Old Days, biochemists) use, if there is enough sample.
Phil
} Hi, All- } } I have a client who would like to figure out the composition (in terms of } protein, carbohydrates, and nucleic acids) of small (0.2um - 1um) abiotic } particles. They have already tried to stain with DAPI and a couple of } other things (I don't know what) and look with fluorescence and flow } cytometry, but the size/signal is too low. } } The only approach I know is microscopy (I'm a one-trick pony). Should they } be pursuing other methods, talking to biochemists, geochemists, food } nutritionists? Their sample size is very tiny; microscopic. } } For microscopy all I can think of is things like antibodies to nucleic } acids, hitting it with all kinds of lectins, and other shotgun approaches. } They have been able to get some of these on a coated grid with an airfuge; } they are pretty electron dense. I shudder to think that I will need to } embed and section them, but I can give it a shot. No idea if they also } contain metallic particles, but I would not be surprised. } } Does anyone have any ideas how to approach this challenge? } } Mahalo (thanks), } 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* } **************************************************************************** -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Fri May 19 09:59:26 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 k4JExOCS005864 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 19 May 2006 09:59:25 -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 k4JFZD4l007714 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 19 May 2006 11:35:13 -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 -- Fri, 19 May 2006 10:59:21 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230904c0938ce13363-at-[141.209.160.132]} 4, 22 -- In-Reply-To: {200605182211.k4IMBkbY028901-at-ns.microscopy.com} 4, 22 -- References: {200605182211.k4IMBkbY028901-at-ns.microscopy.com} 4, 22 -- Date: Fri, 19 May 2006 10:59:20 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] Composition of abiotic particles 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 19 May 2006 14:59:21.0883 (UTC) FILETIME=[CFB4A6B0:01C67B54] 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==============================
Hi Jim, We were apprehensive 3-4 years ago when we made this switch, but were quickly satisfied with the plain-paper work-print quality and speed of an HP Laserjet 4100, cost around $2000 back then. You can always treat yourself to a laser paper of higher brightness, like 98-105, than the noticeably dingy photo-copier paper we find ourselves settling for most of the time because "that's what's loaded in the printer". We "never" make glossies any more; a rare inkjet grayscale run of printing is the nearest thing. Publications are all done via digital files now. A local company that is quoting us on a CCD system for Philips EM 420 says their customers are now very satisfied with
$625 Hewlett Packard LaserJet 2420 MonoChrome Laser Printer (optional purchase) * Up to 30 ppm speed, 1200 dpi. Add $375 for network/duplexing capability.
and they advise: "The printer listed is the printer most recommended for gray-scale images. I can quote whatever printer you specify. However, once you are using digital images, you may find that you don't have a need for printing images. Before deciding that you want a high-quality printer, it would be good to speak with a few colleagues using CCD systems and ask how much printing they do.".
There is a frequent rumor among some experts that HP Laser printers have undocumented capability to produce at 1800 dpi. At any event, we print at something like 120-150 lines per inch; 120 would leave each 1200 dpi pixel sized 10 x 10 dots, giving a range of 10x10 gray levels, but we feel no constraints or shortcomings-- output may not be 256 gray levels, but seems very adequate. We scan negatives routinely at 800 ppi, sometimes 1200 ppi using Epson flatbeds via Photoshop plug-in, and we set up our output-screen properties/ preferences using the interface options between Photoshop and the HP printer that show up under Page Setup and Print windows (Macintosh); this and the grayscale display preferences make differences you probably want to explore and choose among. I'm too preoccupied to dig for our exact numerical settings; they have been unchanged for too long to remember the decisions. we settled on.
-mike reedy-
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-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
On May 18, 2006, at 3:07 PM, tina-at-pbrc.hawaii.edu wrote:
} I have a client who would like to figure out the composition (in terms } of } protein, carbohydrates, and nucleic acids) of small (0.2um - 1um) } abiotic } particles. They have already tried to stain with DAPI and a couple of } other things (I don't know what) and look with fluorescence and flow } cytometry, but the size/signal is too low. } } The only approach I know is microscopy (I'm a one-trick pony). Should } they } be pursuing other methods, talking to biochemists, geochemists, food } nutritionists? Their sample size is very tiny; microscopic. } } For microscopy all I can think of is things like antibodies to nucleic } acids, hitting it with all kinds of lectins, and other shotgun } approaches. } They have been able to get some of these on a coated grid with an } airfuge; } they are pretty electron dense. I shudder to think that I will need to } embed and section them, but I can give it a shot. No idea if they also } contain metallic particles, but I would not be surprised. } } Does anyone have any ideas how to approach this challenge? } Dear Tina, There are some very sensitive methods to measure protein, carbohydrate, and nucleic acids that give quantitative information about composition. If the particles can be dissolved, the protein can be separated from the carbs and NAs by centrifugation, then it can be hydrolysed into amino acids and run through a column to get the composition, or it can be subjected to a Lowry determination (or, perhaps, more up-to-date procedures). Similarly, NA can be amplified by quantitative PCR, then quantitated or sequenced. I'm pretty sure that carbs can also be quantitated biochemically, but I am not at all familiar with those methods. Of course, none of these methods will give any structural info, so if that is needed, EM would be indicated. Talking to several members of a biology department should lead to the experts in dealing with the components of the particles, who would be able to give details on how best to proceed. Metals could be determined and quantitated by atomic absorption spectroscopy, but again this gives only overall composition, not structural info. The small sample size makes it essential that the best methods be identified before proceeding; i.e., doing it right is better than doing it fast. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Fri May 19 11:28:14 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 k4JGSEXI020363 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 19 May 2006 11:28:14 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 92C0A34F1F 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 19 May 2006 09:28:12 -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 id 19FFB34AEF 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 19 May 2006 09:28:10 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200605182207.k4IM7dPu021316-at-ns.microscopy.com} 4, 22 -- References: {200605182207.k4IM7dPu021316-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {937add6f783aaf5fdfce9dea2c131dd6-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Composition of abiotic particles 4, 22 -- Date: Fri, 19 May 2006 09:39:01 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Hi Tina, The University of Georgia Complex Carbohydrate Research Center (CCRC) may be able to help. They have an amazing state-of-the-art facility - I call it "The Palace";-) Check out www.ccrc.uga.edu to see if someone there can help with advice. best, Beth
On Thursday, May 18, 2006, at 06:08 PM, 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 would like to figure out the composition (in terms } of } protein, carbohydrates, and nucleic acids) of small (0.2um - 1um) } abiotic } particles. They have already tried to stain with DAPI and a couple of } other things (I don't know what) and look with fluorescence and flow } cytometry, but the size/signal is too low. } } The only approach I know is microscopy (I'm a one-trick pony). Should } they } be pursuing other methods, talking to biochemists, geochemists, food } nutritionists? Their sample size is very tiny; microscopic. } } For microscopy all I can think of is things like antibodies to nucleic } acids, hitting it with all kinds of lectins, and other shotgun } approaches. } They have been able to get some of these on a coated grid with an } airfuge; } they are pretty electron dense. I shudder to think that I will need to } embed and section them, but I can give it a shot. No idea if they also } contain metallic particles, but I would not be surprised. } } Does anyone have any ideas how to approach this challenge? } } Mahalo (thanks), } 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============================== } 8, 19 -- From tina-at-pbrc.hawaii.edu Thu May 18 17:07:26 2006 } 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu } [128.171.22.7]) } 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4IM7Qns021148 } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 } 17:07:26 -0500 } 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) } 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id } k4IM7Mf6010712 } 8, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 } verify=NO) } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 } 12:07:22 -1000 (HST) } 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu } (8.12.11/8.12.11/Submit) with ESMTP id k4IM7Lu2010709 } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 May 2006 } 12:07:21 -1000 (HST) } 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned } process doing -bs } 8, 19 -- Date: Thu, 18 May 2006 12:07:20 -1000 (HST) } 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } 8, 19 -- X-Sender: tina-at-halia } 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} } 8, 19 -- Subject: Composition of abiotic particles } 8, 19 -- Message-ID: {Pine.GSO.4.21.0605181159010.10514-100000-at-halia} } 8, 19 -- MIME-Version: 1.0 } 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII } ==============================End of - } Headers============================== } ********************************************************************** 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)
We are looking to purchase some software that will log use of the computers that are interface to our microscopes.
Basically on a per user basis the software would record when the user log on and off, and would record uses of the imaging packages installed on the computer.
Currently we do this using the built in Windows 2000/ XP security event logs. This does the job but is not very user friendly.
All systems in question are PCs (no Mac systems) and the users log on with Windows Active Directory accounts.
Does any one out there know of any package that does this? It would be particularly useful if the software would not only record the data but collate it too, i.e. provide tables of total usage per user over a given time period.
Thanks
Lloyd Williams
Manager of Bio-Imaging Core Facility
Hunter College
695 Park Ave
New York NY 10021
==============================Original Headers============================== 18, 20 -- From Williams-at-genectr.hunter.cuny.edu Fri May 19 12:49:00 2006 18, 20 -- Received: from genectr.hunter.cuny.edu (genectr.hunter.cuny.edu [146.95.150.34]) 18, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4JHmxVB012792 18, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 19 May 2006 12:49:00 -0500 18, 20 -- Content-class: urn:content-classes:message 18, 20 -- MIME-Version: 1.0 18, 20 -- Content-Type: text/plain; 18, 20 -- charset="us-ascii" 18, 20 -- Subject: Software to Log Use of a Computer Interfaced to a Microscope 18, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 18, 20 -- Date: Fri, 19 May 2006 13:50:09 -0400 18, 20 -- Message-ID: {DD9D9EF525BDB444A46962D3D4F6E9A5011555FE-at-xchange2.bio.hunter.cuny.edu} 18, 20 -- X-MS-Has-Attach: 18, 20 -- X-MS-TNEF-Correlator: 18, 20 -- Thread-Topic: Software to Log Use of a Computer Interfaced to a Microscope 18, 20 -- Thread-Index: AcZ7XJDD2rA9fA3BSgmQrMnFXtGeIw== 18, 20 -- From: "Lloyd Williams" {Williams-at-genectr.hunter.cuny.edu} 18, 20 -- To: {Microscopy-at-microscopy.com} 18, 20 -- Content-Transfer-Encoding: 8bit 18, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4JHmxVB012792 ==============================End of - Headers==============================
There are two potential problems with the approach you describe. One is that the copper grids, as pointed out in another reply, may be reacting to the salts in the growth medium. He other problem is that the bacteria are in th eprocess of forming a biofilm on the support film surface during the incubation time.
If you only want to examine the bacteria by negative stain, you can place a drop of the culture on the support film surface, leave for about 30 sec and then replace the drop with a drop of negative stain. You have to be careful to make sure there are no salts in the growth medium that could cause the negative stain, and in particular the uranyl acetate, to precipitate. There is no need to centrifuge the bacteria because there are usually enough in the drop to fall to the surface.
As with most techniques, there are many variations on this simple method, but it is always a good policy to try the easier way first to make sure it is possible to obtain results. The next easiest way is to deposit a carbon film onto a freshly-cleaved mica surface and then allow a drop of bacterial suspension to infiltrate between the carbon film and the mica. If you then place a clean grid on the carbon film you should be able to pick it up onto the support grid and then negative stain. The advantage of this method is that you don't have to worry about the support film being hydrophilic - it already is. The disadvantage is that the carbon film is very fragile so needs a small mesh grid to support it. Even better support comes form holey grids.
If you are interested in examining the bacteria as they grow on the grids, first switch to gold or nickel grids. You may then want to extensively wash the bacteria before you stain them to remove all the extracellular material they secrete in the short time they have been incubated. You can check to see if what I am saying is correct by taking a grid with bacteria on directly from the culture medium (after incubation) and plunge-freezing it in liquid propane. From there you freeze substitute it in ethanol (with a little osmium tetroxide if you wish), warm it slowly to 4 degrees and then critical point dry it and look at it in the SEM. My guess is that the bacteria will be almost invisible because they will be covered in a large amount of slimy looking stuff.
I hope this helps.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
} From: {Randi.Olsen-at-fagmed.uit.no} } Reply-To: {Randi.Olsen-at-fagmed.uit.no} } Date: Fri, 19 May 2006 02:47:23 -0500 } To: {pwebster-at-hei.org} } Subject: [Microscopy] FW: NEGATIVE STAINING ENTEROCOCCI } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } } } _____________________________________________ } X-from: Randi Olsen } Sent: 18. mai 2006 16:04 } To: 'Microscopy-at-microscopy.com' } Subject: NEGATIVE STAINING ENTEROCOCCI } } Dear all, } We have problems with negative staining of E. Coli, and have tried two } different approaches with both Uranyl acetate and PTA. } Without being able to detect the flagellas, we see partly collapsed } bacterias and lots of dirt (growing bacterias directly on the grids) } We've also tried embed the E.coli in a mixture of methylcellulose and } uranyl acetate without any luck. } We've used carboncoated formvar films on copper grids. } I enclose the procedure for preparing cells grown on grids, observations } and some questions form the scientist in charge of this project. } We might be wrong trying to grow cells directly on grids? } } } Our aim is to prepare grids for TEM with a suitable cell number for } reliable anaysis without damaging the cells by centrifugation. } We plan to grow the cells directly onto the grids and have performed the } following: } } Inoculate cultures of enterococci in TSB+0.25%glucose and grow overnight } at 37*C without shaking. (TSB is a bacterial growth medium made from } casein and soya peptone). } Dilute cultures 100-fold in 10 ml growth medium in microtiterplate wells } containing TEM grids (should contain ca.107 cells/ml). } Incubate at 37*C without shaking for 1-4 hours. } Pick up grids at different time-points (i.e. 1h, 2h, 3h and 4h) from the } culture. } Dip the grid in particle free water. } Stain the cells with 0.5% uranyl acetate for 30". } Rinse the grids gently in particle free water before air-drying them for } 10-15'. } } Observations made: } 1. there are low cell-numbers on the grids } 2. there's a lot of stained debris or precipitate on the grids and it } seems to increase during the incubation. } } Questions: } 1. What cell density is required to obtain one layer of cells on the } grid but not cells on top of each other? } 2. Are there ways of increasing the bacterial affinity for the grids? } 3. Are there ways to reduce the amount of debris on the grids? } 4. Can grids be incubated on top of a nitrocellulose filter without } beeing damaged? } } } } } Best regards } } Randi Olsen } Department of Electron Microscopy } University of Tromso } Norway } } } ==============================Original Headers============================== } 15, 25 -- From Randi.Olsen-at-fagmed.uit.no Fri May 19 02:42:19 2006 } 15, 25 -- Received: from mux2.uit.no (mux2.uit.no [129.242.5.252]) } 15, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4J7gIuH019283 } 15, 25 -- for {microscopy-at-microscopy.com} ; Fri, 19 May 2006 02:42:19 -0500 } 15, 25 -- Received: from eden5.ad.uit.no (eden5.ad.uit.no [129.242.8.11]) } 15, 25 -- by mux2.uit.no (8.13.6/8.13.6/Mux) with ESMTP id k4J7fx6R032043 } 15, 25 -- for {microscopy-at-microscopy.com} ; Fri, 19 May 2006 09:42:18 +0200 } (CEST) } 15, 25 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 } 15, 25 -- Content-class: urn:content-classes:message } 15, 25 -- MIME-Version: 1.0 } 15, 25 -- Content-Type: text/plain; } 15, 25 -- charset="us-ascii" } 15, 25 -- Subject: FW: NEGATIVE STAINING ENTEROCOCCI } 15, 25 -- Date: Fri, 19 May 2006 09:42:13 +0200 } 15, 25 -- Message-ID: {6B3D9BCAC4A0F941AC7625C4589F61FC4F7EFB-at-eden5.ad.uit.no} } 15, 25 -- X-MS-Has-Attach: } 15, 25 -- X-MS-TNEF-Correlator: } 15, 25 -- Thread-Topic: NEGATIVE STAINING ENTEROCOCCI } 15, 25 -- thread-index: AcZ6bAOqp+ckv3asTc6wZn588yH8jgAFZmYgACWGKyA= } 15, 25 -- From: "Randi Olsen" {Randi.Olsen-at-fagmed.uit.no} } 15, 25 -- To: {microscopy-at-microscopy.com} } 15, 25 -- X-Virus-Scanned: : ok } 15, 25 -- X-Scanned-By: MIMEDefang 2.56 on 129.242.5.252 } 15, 25 -- Content-Transfer-Encoding: 8bit } 15, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k4J7gIuH019283 } ==============================End of - Headers==============================
==============================Original Headers============================== 12, 19 -- From PWebster-at-hei.org Fri May 19 13:54:08 2006 12, 19 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 12, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4JIs7kk027881 12, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 19 May 2006 13:54:07 -0500 12, 19 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 12, 19 -- Fri, 19 May 2006 18:54:07 +0000 12, 19 -- User-Agent: Microsoft-Entourage/11.2.3.060209 12, 19 -- Date: Fri, 19 May 2006 11:54:06 -0700 12, 19 -- Subject: Re: [Microscopy] FW: NEGATIVE STAINING ENTEROCOCCI 12, 19 -- From: "Webster, Paul" {PWebster-at-hei.org} 12, 19 -- To: {Randi.Olsen-at-fagmed.uit.no} , {Microscopy-at-microscopy.com} 12, 19 -- Message-ID: {C093625E.A305%PWebster-at-hei.org} 12, 19 -- Thread-Topic: [Microscopy] FW: NEGATIVE STAINING ENTEROCOCCI 12, 19 -- Thread-Index: AcZ7dZp+2UNP6OdoEdqZigANk7Zh7g== 12, 19 -- In-Reply-To: {200605190747.k4J7lNO9026705-at-ns.microscopy.com} 12, 19 -- Mime-version: 1.0 12, 19 -- Content-type: text/plain; 12, 19 -- charset="US-ASCII" 12, 19 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
On May 11, 2006, at 1:13 PM, dlowry-at-asu.edu wrote:
} Colleagues, } } I am writing to get information on web-pages which have compiled links } to bio-imaging and microscopy facilities both within the US and } internationally. } } I have searched and located a few such web-sites, but they are not } very comprehensive and many of the links are dead. } } If anyone in the community may have web-addresses or information on } sites of this nature, I would appreciate their input. } } Thank you, } } David Lowry } School of Life Sciences } Arizona State University } Tempe, AZ 85287-4501 } office: 480-727-0725 } lab: 480-965-2463 } Dear David, Our lab web page has links to our work. We do not have links to other labs, AFAIK. The URL is
http://www.jensenlab.caltech.edu/
Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 6, 23 -- From tivol-at-caltech.edu Fri May 19 17:59:11 2006 6, 23 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4JMxBhR009918 6, 23 -- for {microscopy-at-msa.microscopy.com} ; Fri, 19 May 2006 17:59:11 -0500 6, 23 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 6, 23 -- by water-ox-postvirus (Postfix) with ESMTP 6, 23 -- id 1B342351A7; Fri, 19 May 2006 15:59:11 -0700 (PDT) 6, 23 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 6, 23 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP 6, 23 -- id DB6683673C; Fri, 19 May 2006 15:59:08 -0700 (PDT) 6, 23 -- In-Reply-To: {200605112013.k4BKDlgT025208-at-ns.microscopy.com} 6, 23 -- References: {200605112013.k4BKDlgT025208-at-ns.microscopy.com} 6, 23 -- Mime-Version: 1.0 (Apple Message framework v624) 6, 23 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 6, 23 -- Message-Id: {3f06ad4e55427d9f2192974a06ccac69-at-caltech.edu} 6, 23 -- Content-Transfer-Encoding: 7bit 6, 23 -- Cc: microscopy-at-msa.microscopy.com, Grant Jensen {jensen-at-caltech.edu} 6, 23 -- From: Bill Tivol {tivol-at-caltech.edu} 6, 23 -- Subject: Re: [Microscopy] 6, 23 -- Date: Fri, 19 May 2006 16:10:00 -0700 6, 23 -- To: dlowry-at-asu.edu 6, 23 -- X-Mailer: Apple Mail (2.624) 6, 23 -- 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 bxp-at-cfdrc.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: bxp-at-cfdrc.com Name: Prabhakar Pandian
Title-Subject: [Filtered] X-cite vs Lamda LS vs Hg Lamp
Question: Hello, We are in the market for adding a fluorescent set-up for our microscope and are looking at the X-cite 120 and the Lambda LS from Sutter Instruments. Can anyone provide pros and cons perspective.
We already have a Hg Burner for another microscope and know that is the best for the UV range.
Hello Randi, I do not have answers for all the questions but this is what we have done in the past while negatively staining Gram negative bacteria Questions: } } 1. What cell density is required to obtain one layer of cells on the } } grid but not cells on top of each other? 2. Are there ways of increasing the bacterial affinity for the grids? } } 3. Are there ways to reduce the amount of debris on the grids? } } 4. Can grids be incubated on top of a nitrocellulose filter without } } beeing damaged?
Answer: It might be challenging to achieve a monolayer of bacterial cells on the grid as every bacteria behaves a little differently when it comes to attaching to any surface. In order to view flagella or pili, we grew the bacteria in stationary cultures to prevent mechanical loss of flagella or pili. Once the bacteria were in in their log phase we pipetted out around 100 microliters of the culture and floated a fomvar coated nickle grid on it for about 5 mins and then let the grids dry. We found the incubation time to vary with different strains. These grids were rinsed in distilled water for 30 sec to a min. I have found that rinsing the grids was very important to remove all the debris and residual media from the grid leaving a clean prep. If you find low number of bacterial cells you can try increasing the incubation time and decreasing the rinse time. Staining: The grids were stained for 10 mins with 0.2% UA for 10 mins followed by a 1 min rinse with distilled water. The grids were then air dried before examination.
I found rinsing steps were key to a good prep. You have to play with the innoculation and rinsing steps to obtain desirable results.
Hope that helps. Good luck with your experiments.
regards, Vinod Nair Electron microscopy lab New Mexico State University Las Cruces NM 88003
} } 2. Are there ways of increasing the bacterial affinity for the grids? } } 3. Are there ways to reduce the amount of debris on the grids? } } 4. Can grids be incubated on top of a nitrocellulose filter without } } beeing damaged?
On 5/19/06, PWebster-at-hei.org {PWebster-at-hei.org} wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi Randi, } } There are two potential problems with the approach you describe. One is that } the copper grids, as pointed out in another reply, may be reacting to the } salts in the growth medium. He other problem is that the bacteria are in th } eprocess of forming a biofilm on the support film surface during the } incubation time. } } If you only want to examine the bacteria by negative stain, you can place a } drop of the culture on the support film surface, leave for about 30 sec and } then replace the drop with a drop of negative stain. You have to be careful } to make sure there are no salts in the growth medium that could cause the } negative stain, and in particular the uranyl acetate, to precipitate. There } is no need to centrifuge the bacteria because there are usually enough in } the drop to fall to the surface. } } As with most techniques, there are many variations on this simple method, } but it is always a good policy to try the easier way first to make sure it } is possible to obtain results. The next easiest way is to deposit a carbon } film onto a freshly-cleaved mica surface and then allow a drop of bacterial } suspension to infiltrate between the carbon film and the mica. If you then } place a clean grid on the carbon film you should be able to pick it up onto } the support grid and then negative stain. The advantage of this method is } that you don't have to worry about the support film being hydrophilic - it } already is. The disadvantage is that the carbon film is very fragile so } needs a small mesh grid to support it. Even better support comes form holey } grids. } } If you are interested in examining the bacteria as they grow on the grids, } first switch to gold or nickel grids. You may then want to extensively wash } the bacteria before you stain them to remove all the extracellular material } they secrete in the short time they have been incubated. You can check to } see if what I am saying is correct by taking a grid with bacteria on } directly from the culture medium (after incubation) and plunge-freezing it } in liquid propane. From there you freeze substitute it in ethanol (with a } little osmium tetroxide if you wish), warm it slowly to 4 degrees and then } critical point dry it and look at it in the SEM. My guess is that the } bacteria will be almost invisible because they will be covered in a large } amount of slimy looking stuff. } } I hope this helps. } } Paul Webster. } } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } (213) 273 8026 } pwebster-at-hei.org } } } } From: {Randi.Olsen-at-fagmed.uit.no} } } Reply-To: {Randi.Olsen-at-fagmed.uit.no} } } Date: Fri, 19 May 2006 02:47:23 -0500 } } To: {pwebster-at-hei.org} } } Subject: [Microscopy] FW: NEGATIVE STAINING ENTEROCOCCI } } } } } } } } } } ---------------------------------------------------------------------------- } } 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 } } ---------------------------------------------------------------------------- } } } } } } } } _____________________________________________ } } X-from: Randi Olsen } } Sent: 18. mai 2006 16:04 } } To: 'Microscopy-at-microscopy.com' } } Subject: NEGATIVE STAINING ENTEROCOCCI } } } } Dear all, } } We have problems with negative staining of E. Coli, and have tried two } } different approaches with both Uranyl acetate and PTA. } } Without being able to detect the flagellas, we see partly collapsed } } bacterias and lots of dirt (growing bacterias directly on the grids) } } We've also tried embed the E.coli in a mixture of methylcellulose and } } uranyl acetate without any luck. } } We've used carboncoated formvar films on copper grids. } } I enclose the procedure for preparing cells grown on grids, observations } } and some questions form the scientist in charge of this project. } } We might be wrong trying to grow cells directly on grids? } } } } } } Our aim is to prepare grids for TEM with a suitable cell number for } } reliable anaysis without damaging the cells by centrifugation. } } We plan to grow the cells directly onto the grids and have performed the } } following: } } } } Inoculate cultures of enterococci in TSB+0.25%glucose and grow overnight } } at 37*C without shaking. (TSB is a bacterial growth medium made from } } casein and soya peptone). } } Dilute cultures 100-fold in 10 ml growth medium in microtiterplate wells } } containing TEM grids (should contain ca.107 cells/ml). } } Incubate at 37*C without shaking for 1-4 hours. } } Pick up grids at different time-points (i.e. 1h, 2h, 3h and 4h) from the } } culture. } } Dip the grid in particle free water. } } Stain the cells with 0.5% uranyl acetate for 30". } } Rinse the grids gently in particle free water before air-drying them for } } 10-15'. } } } } Observations made: } } 1. there are low cell-numbers on the grids } } 2. there's a lot of stained debris or precipitate on the grids and it } } seems to increase during the incubation. } } } } Questions: } } 1. What cell density is required to obtain one layer of cells on the } } grid but not cells on top of each other? } } 2. Are there ways of increasing the bacterial affinity for the grids? } } 3. Are there ways to reduce the amount of debris on the grids? } } 4. Can grids be incubated on top of a nitrocellulose filter without } } beeing damaged? } } } } } } } } } } Best regards } } } } Randi Olsen } } Department of Electron Microscopy } } University of Tromso } } Norway } } } } } } ==============================Original Headers============================== } } 15, 25 -- From Randi.Olsen-at-fagmed.uit.no Fri May 19 02:42:19 2006 } } 15, 25 -- Received: from mux2.uit.no (mux2.uit.no [129.242.5.252]) } } 15, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } k4J7gIuH019283 } } 15, 25 -- for {microscopy-at-microscopy.com} ; Fri, 19 May 2006 02:42:19 -0500 } } 15, 25 -- Received: from eden5.ad.uit.no (eden5.ad.uit.no [129.242.8.11]) } } 15, 25 -- by mux2.uit.no (8.13.6/8.13.6/Mux) with ESMTP id k4J7fx6R032043 } } 15, 25 -- for {microscopy-at-microscopy.com} ; Fri, 19 May 2006 09:42:18 +0200 } } (CEST) } } 15, 25 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 } } 15, 25 -- Content-class: urn:content-classes:message } } 15, 25 -- MIME-Version: 1.0 } } 15, 25 -- Content-Type: text/plain; } } 15, 25 -- charset="us-ascii" } } 15, 25 -- Subject: FW: NEGATIVE STAINING ENTEROCOCCI } } 15, 25 -- Date: Fri, 19 May 2006 09:42:13 +0200 } } 15, 25 -- Message-ID: {6B3D9BCAC4A0F941AC7625C4589F61FC4F7EFB-at-eden5.ad.uit.no} } } 15, 25 -- X-MS-Has-Attach: } } 15, 25 -- X-MS-TNEF-Correlator: } } 15, 25 -- Thread-Topic: NEGATIVE STAINING ENTEROCOCCI } } 15, 25 -- thread-index: AcZ6bAOqp+ckv3asTc6wZn588yH8jgAFZmYgACWGKyA= } } 15, 25 -- From: "Randi Olsen" {Randi.Olsen-at-fagmed.uit.no} } } 15, 25 -- To: {microscopy-at-microscopy.com} } } 15, 25 -- X-Virus-Scanned: : ok } } 15, 25 -- X-Scanned-By: MIMEDefang 2.56 on 129.242.5.252 } } 15, 25 -- Content-Transfer-Encoding: 8bit } } 15, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id k4J7gIuH019283 } } ==============================End of - Headers============================== } } } ==============================Original Headers============================== } 12, 19 -- From PWebster-at-hei.org Fri May 19 13:54:08 2006 } 12, 19 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) } 12, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4JIs7kk027881 } 12, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 19 May 2006 13:54:07 -0500 } 12, 19 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; } 12, 19 -- Fri, 19 May 2006 18:54:07 +0000 } 12, 19 -- User-Agent: Microsoft-Entourage/11.2.3.060209 } 12, 19 -- Date: Fri, 19 May 2006 11:54:06 -0700 } 12, 19 -- Subject: Re: [Microscopy] FW: NEGATIVE STAINING ENTEROCOCCI } 12, 19 -- From: "Webster, Paul" {PWebster-at-hei.org} } 12, 19 -- To: {Randi.Olsen-at-fagmed.uit.no} , {Microscopy-at-microscopy.com} } 12, 19 -- Message-ID: {C093625E.A305%PWebster-at-hei.org} } 12, 19 -- Thread-Topic: [Microscopy] FW: NEGATIVE STAINING ENTEROCOCCI } 12, 19 -- Thread-Index: AcZ7dZp+2UNP6OdoEdqZigANk7Zh7g== } 12, 19 -- In-Reply-To: {200605190747.k4J7lNO9026705-at-ns.microscopy.com} } 12, 19 -- Mime-version: 1.0 } 12, 19 -- Content-type: text/plain; } 12, 19 -- charset="US-ASCII" } 12, 19 -- Content-transfer-encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 10, 25 -- From nairvinods-at-gmail.com Sun May 21 23:59:47 2006 10, 25 -- Received: from nz-out-0102.google.com (nz-out-0102.google.com [64.233.162.206]) 10, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4M4xkVV017082 10, 25 -- for {Microscopy-at-microscopy.com} ; Sun, 21 May 2006 23:59:47 -0500 10, 25 -- Received: by nz-out-0102.google.com with SMTP id i1so1000226nzh 10, 25 -- for {Microscopy-at-microscopy.com} ; Sun, 21 May 2006 21:59:46 -0700 (PDT) 10, 25 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 25 -- s=beta; d=gmail.com; 10, 25 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 10, 25 -- b=Z+ISzyssxwFBqVMGkTX36n5RAWR59JeXCwZOifwvh0rWt7DhaC/omOQ43tubA5V/5jAKuPnLFtX/i3l1qVKo/U2AJzIFQ+O8TdfJQobYRMXiItNRmtU6xsB1xpW57ciQjCK4MfH0Igrf6a5WM3TFsEUMugPmChUF24FmSPrE9uc= 10, 25 -- Received: by 10.64.203.16 with SMTP id a16mr419527qbg; 10, 25 -- Sun, 21 May 2006 21:59:46 -0700 (PDT) 10, 25 -- Received: by 10.64.201.11 with HTTP; Sun, 21 May 2006 21:59:46 -0700 (PDT) 10, 25 -- Message-ID: {ea42a3900605212159g74e3c6f9qf322d4702b8f20f-at-mail.gmail.com} 10, 25 -- Date: Sun, 21 May 2006 22:59:46 -0600 10, 25 -- From: "Vinod Nair" {nairvinods-at-gmail.com} 10, 25 -- To: Microscopy-at-microscopy.com 10, 25 -- Subject: Re: [Microscopy] Re: FW: NEGATIVE STAINING ENTEROCOCCI 10, 25 -- In-Reply-To: {200605191859.k4JIx37l002959-at-ns.microscopy.com} 10, 25 -- MIME-Version: 1.0 10, 25 -- Content-Type: text/plain; charset=UTF-8; format=flowed 10, 25 -- Content-Disposition: inline 10, 25 -- References: {200605191859.k4JIx37l002959-at-ns.microscopy.com} 10, 25 -- Content-Transfer-Encoding: 8bit 10, 25 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k4M4xkVV017082 ==============================End of - Headers==============================
Thanks to all for the advice on sectioning JB 4 resin, with its tendency to fold and curl. I am summarizing the replies below:
1) From Glen McDonald: "For serial sections, which thankfully I haven't had to do with JB-4 in many, many years, I got a tackle box, found in any small parts supplier or fishing supply shop - a clear plastic box with 2x6 or 4x6 array of compartments about 2 inches square. Fill with deionized water or 3% ethanol. As the section comes off of the knife edge, either lay an eyelash across the bottom edge to prevent the curling, or grab with a pair of Dumont #55 forceps. move the eyelash or forceps along with the motion of the section, then lift the section and drop onto the liquid. Place one section in each compartment to fill the tackle box, then mount them by immersing the slide and bringing it up under the section. *Gently* touch one corner of the section to guide into position. If not gently enough, the section will cling to the eyelash and wad up like a used tissue during a bad cold." There were a couple other variations on this technique of lifting slides up underneath the sections. Off to the Bass Pro Shops electron microscopy department for me!
2) Several people described helping the section come off the block by pulling with a forceps on one corner during the cutting stroke or holding it flat with an eyelash or brush, then flicking the section quickly onto a drop of water or water/ethanol mixture. Dexterity required, methinks.
3) Another repeated suggestion was to put the sections onto drops of water with a little ammonium hydroxide in it.
4) As mentioned above, putting sections into water with ethanol, up to 50%, was mentioned several times, sometimes followed by transferring sections to distilled water afterwards.
5) Don't use JB 4. (My favorite.)
6) Tobias Baskin has published his own formulation of BMM resin which apparently sections much better. He uses DTT in the mix and says he is happy to help anyone with this resin. We haven't tried this, yet, but we intend to. Tobias posts to the list often, but if you can't find his email, let me know and I'll forward your questions to him.
7) Humidity should be in the 40-50% range and the block should neither be too wet or too dry or "sectioning is nearly impossible". From Ralph Common. Humidity? In Missouri? Who could have guessed?
8) Related to 7, if the block is too soft, it won't cut well. This one did it for us! Four more hours in the oven solved the worst of the problem. Sections coming off flat and staying flat. These sections did fold when placed on 50% ethanol/water, but did not fold when placed on distilled water. Yaaaaaayy! Thanks, Teri Johnson!!
This was a crash course in JB 4 emergency microtomy. Our client is happy, and Cheryl is tired. Cheryl gets a free lunch at the restaurant of her choice (in Columbia, anyway).
Thanks again to all who responded. You guys are great!
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============================== 16, 24 -- From TindallR-at-missouri.edu Mon May 22 09:08:06 2006 16, 24 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4ME86Yd017418 16, 24 -- for {microscopy-at-microscopy.com} ; Mon, 22 May 2006 09:08:06 -0500 16, 24 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 16, 24 -- Mon, 22 May 2006 09:08:05 -0500 16, 24 -- Content-class: urn:content-classes:message 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-Type: text/plain; 16, 24 -- charset="us-ascii" 16, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 24 -- Subject: JB 4 sections: Summary (long) 16, 24 -- Date: Mon, 22 May 2006 09:08:05 -0500 16, 24 -- Message-ID: {BA876152E8653240BE8572E897083EE701849FEE-at-UM-EMAIL09.um.umsystem.edu} 16, 24 -- X-MS-Has-Attach: 16, 24 -- X-MS-TNEF-Correlator: 16, 24 -- Thread-Topic: JB 4 sections: Summary (long) 16, 24 -- Thread-Index: AcZ9qSVYx+R/zjlKQ7+NK+p6LgTVXA== 16, 24 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 16, 24 -- To: {microscopy-at-microscopy.com} 16, 24 -- Cc: "Jensen, Cheryl A." {JensenC-at-missouri.edu} 16, 24 -- X-OriginalArrivalTime: 22 May 2006 14:08:05.0976 (UTC) FILETIME=[258FBD80:01C67DA9] 16, 24 -- Content-Transfer-Encoding: 8bit 16, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4ME86Yd017418 ==============================End of - Headers==============================
We are seeking a biological scientist with experience in electron microscopy and other imaging techniques to direct the Rothamsted Centre for Bioimaging. This state of the art facility opened in 2003 and houses 3 electron microscopes, a confocal microscope, digital imaging and associated image analysis software, a laser capture system and a fully equipped cryo-preparation laboratory. The centre provides a high quality service for a wide range of users both inside and outside the research Institute. The Institute is the largest agricultural research centre in the United Kingdom. The scientific research ranges from studies of genetics, biochemistry, cell biology and soil processes to investigations at the ecosystem and landscape scale.
You will be responsible for developing innovative research projects in collaboration with other staff and for providing scientific and technical advice on bioimaging within institute research programmes. You will need to keep abreast of technologies and instrumentation to ensure that the Centre develops new science and imaging applications and remains at the cutting edge. The post also entails management of two support staff.
Applicants should have a PhD or other higher qualification in a relevant biological science, plus at least 5-7 years' postdoctoral experience in microscopy, imaging or structural biology. You must be well versed in both conventional and cryo-preparative techniques and their application to biological samples. Experience in running a microscopy facility would be an advantage.
The appointment will be at Band 5 with a starting salary normally within the range Ł27,584 - Ł32,122 per annum.
For further information please contact Professor John Lucas ++ 44(0)1582 763133 x 2779 or john.lucas-at-bbsrc.ac.uk).
Apply by application form only, please visit our website http://www.rothamsted.bbsrc.ac.uk/careers/vacancies/Vacancies.html for further information and details regarding the application process.
Closing date: 8 June 2006
** Dr Smita Kurup CPI Division Rothamsted Research Harpenden Herts AL5 2JQ
Tel No. 01582 763133 ext 2589 Fax No. 01582 763010
==============================Original Headers============================== 14, 20 -- From jrunions-at-brookes.ac.uk Mon May 22 12:39:33 2006 14, 20 -- Received: from brookes.ac.uk (csmail1.brookes.ac.uk [161.73.1.23]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4MHdXKn001023 14, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 22 May 2006 12:39:33 -0500 14, 20 -- Received: from [161.73.107.141] (bc8bf582.brookes.ac.uk [161.73.107.141]) 14, 20 -- by brookes.ac.uk (8.13.6/8.13.6) with ESMTP id k4MHcOkU016838 14, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 22 May 2006 18:38:25 +0100 (BST) 14, 20 -- Message-ID: {4471F712.80200-at-brookes.ac.uk} 14, 20 -- Date: Mon, 22 May 2006 18:38:26 +0100 14, 20 -- From: John Runions {jrunions-at-brookes.ac.uk} 14, 20 -- User-Agent: Mozilla Thunderbird 1.0 (Windows/20041206) 14, 20 -- X-Accept-Language: en-us, en 14, 20 -- MIME-Version: 1.0 14, 20 -- To: Microscopy Listserver {Microscopy-at-Microscopy.Com} 14, 20 -- Subject: Position announcement - Head of Imaging Facility 14, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 14, 20 -- Content-Transfer-Encoding: 8bit 14, 20 -- X-MailScanner-Information: Oxford Brookes University MailScanner 14, 20 -- X-MailScanner: Clean 14, 20 -- X-MailScanner-From: jrunions-at-brookes.ac.uk ==============================End of - Headers==============================
Hello, Does anyone have some experience with the: Balzers RES 010 Rapid Etching System I was wondering if it could be used for coating of samples for SEM as well as etching?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Mon May 22 12:49:16 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 k4MHnG5k011110 2, 24 -- for {microscopy-at-microscopy.com} ; Mon, 22 May 2006 12:49:16 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 196CFC1E61 2, 24 -- for {microscopy-at-microscopy.com} ; Mon, 22 May 2006 10:49:16 -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 22888-07 for {microscopy-at-microscopy.com} ; 2, 24 -- Mon, 22 May 2006 10:49:10 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id 65C59C1E59; Mon, 22 May 2006 10:49:10 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 40392C1E3E 2, 24 -- for {Microscopy-at-Microscopy.Com} ; Mon, 22 May 2006 10:49:10 -0700 (PDT) 2, 24 -- Date: Mon, 22 May 2006 10:49:09 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: Microscopy-at-microscopy.com 2, 24 -- Subject: Balzers RES 010 Rapid Etching System 2, 24 -- Message-ID: {Pine.SOC.4.64.0605221048210.18594-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==============================
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 roderi_co-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: roderi_co-at-yahoo.com Name: Roman Derico
Organization: Faculty of food technology and biotechnology
Title-Subject: [Filtered] TEM - Problem with heating specimen holder
Question: Hi! I have a weird problem. I have two heating specimen holders and recently I tried to use them on Jeol JEM200CX TEM (both holders are original parts from Jeol; last time they have been used on this microscope several years ago). When I inserted the holder in microscope, the electron beam disappeared from screen. I moved the holder step-by-step from position I to position II and changed the x and y position systematically (with specimen shifting knobs), but no beam appeared on screen. The beam doesn't appear on screen even when heating holder is empty (with specimen grid removed from it). The strange thing is that hole on heating holder matches exactly in position with holes on specimen holders of other types (it corresponds to specimen position I hole), and all other types of specimen holders work well. Does anyone have idea why beam doesn't pass through heating holder?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dfine-at-seton.org) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, May 22, 2006 at 16:28:41 ---------------------------------------------------------------------------
Email: dfine-at-seton.org Name: Belinda Torres
Organization: University of Texas
Education: Graduate College
Location: Austin,Texas
Question: I am doing some experiments in electron microscopy and I am having a hard time removing wrinkles from my resin(thicks) sections. Can you help.
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 jacqui.ross-at-auckland.ac.nz as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: jacqui.ross-at-auckland.ac.nz Name: Jacqueline Ross
Organization: The University of Auckland
Title-Subject: [Filtered] Job posting: Lecturer position in Auckland, New Zealand
Question: The Department of Anatomy with Radiology has a vacancy as detailed below. This person will also be associated with the Biomedical Imaging Research Unit. Please see the link to the job description if you are interested.
LECTURER
Anatomy with Radiology
School of Medical Sciences
Faculty of Medical and Health Sciences
The University of Auckland
Vacancy Number A304-06
We require a Lecturer to facilitate imaging research in cellular and molecular imaging and assist in providing for the ongoing development of imaging technologies within the Biomedical Imaging Research Unit and the wider faculty. The appointee will also be expected to provide leadership to both undergraduate and postgraduate teaching programmes, to assist in the development, planning and management of courses and programmes relevant to their field of expertise and to act in a supervisory role to graduate students.
Applications are invited from PhD qualified candidates with a track record of research and teaching in the areas of cellular/molecular imaging and cell and tissue biology.
For further information and to apply online, please visit http://www.vacancies.auckland.ac.nz/ or alternatively call +64 9-373 7599 ext 83000.
Please quote the vacancy number. Applications close 23 June 2006.
The University has an equal opportunities policy and welcomes opportunities policy and welcomes applications from all qualified persons.
I would suggest that you post the details for "foreign" applicants as they are not likely to be acceptable to your solicitation.
There are restrictions on US jobs as well on non-US jobs. I think that each solicitation should identify the nuances of their particular geopolitical/geographic location.
That might save many folks a lot of wasted time. Just MO.
gary g.
At 07:24 PM 5/22/2006, you wrote:
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==============================Original Headers============================== 12, 21 -- From gary-at-gaugler.com Mon May 22 21:46:50 2006 12, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4N2knqd030454 12, 21 -- for {microscopy-at-microscopy.com} ; Mon, 22 May 2006 21:46:49 -0500 12, 21 -- Received: (qmail 25679 invoked from network); 22 May 2006 19:46:47 -0700 12, 21 -- Received: by simscan 1.1.0 ppid: 25676, pid: 25677, t: 0.1906s 12, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 21 -- by qsmtp3 with SMTP; 22 May 2006 19:46:47 -0700 12, 21 -- Message-Id: {7.0.1.0.2.20060522194211.023d1c28-at-gaugler.com} 12, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 21 -- Date: Mon, 22 May 2006 19:46:49 -0700 12, 21 -- To: jacqui.ross-at-auckland.ac.nz 12, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 21 -- Subject: Re: [Microscopy] viaWWW: Job posting: Lecturer position in 12, 21 -- Auckland, New Zealand 12, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 21 -- In-Reply-To: {200605230224.k4N2OG9j023317-at-ns.microscopy.com} 12, 21 -- References: {200605230224.k4N2OG9j023317-at-ns.microscopy.com} 12, 21 -- Mime-Version: 1.0 12, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Okay, I'm a biologist and have absolutely no idea of how to go about setting up a Philips CM-10 for dark field imaging. Now I have a user who needs it and we need some help.
I would appreciate some suggestions for basic settings to get us started such as aperture sizes, spot size, tilt angles, etc.
Thanks in advance, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 6, 21 -- From dsherman-at-purdue.edu Mon May 22 22:07:26 2006 6, 21 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4N37QYM008244 6, 21 -- for {microscopy-at-microscopy.com} ; Mon, 22 May 2006 22:07:26 -0500 6, 21 -- Received: from exchange.purdue.edu ([128.210.63.234]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Mon, 22 May 2006 23:07:39 -0400 6, 21 -- Received: from 12.208.56.23 ([12.208.56.23]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 6, 21 -- Tue, 23 May 2006 03:07:27 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.2.3.060209 6, 21 -- Date: Mon, 22 May 2006 23:07:26 -0400 6, 21 -- Subject: Dark field TEM 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C097F4AE.2A94%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Dark field TEM 6, 21 -- Thread-Index: AcZ+FgS1Qwy8kuoJEdqGcAAKlcoUxg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 23 May 2006 03:07:39.0844 (UTC) FILETIME=[0CF59840:01C67E16] ==============================End of - Headers==============================
Hi, I think that operation in dark-field (DF) mode on CM10 is similar to CM12.
Simply, we are using following procedure for beginners in DF on our CM12: -start with well aligned microscope, C2 aperture 100-200 µm, spot size 2, 80kV -insert suitable sample (it should be thin!! e.g. DNA on carbon support film) -adjust eucentric height of the sample -check rotation center and pivot points in bright-field mode -check stigmators -switch to diffraction mode and center diffraction spot -insert and center OBJ aperture -press DF button on CM10 panel -press RESET button under OPCON to center diffraction spot in DF mode -move the diff. spot with Multifunction knobs just behind the rim of the aperture disk on main screen -switch to imaging mode and you should see dark-field image of your sample -focus the sample and adjust illumination with Intensity button
You should try different OBJ apertures to get optimal image in DF with your sample. I hope the procedure will work for you.
Best regards from Prague Oldrich
----------------------------------- Oldrich Benada Institute of Microbiology Acad. Sci CR Videnska 1083 142 20 Prague 4 Czech Republic
} } } ---------------------------------------------------------------------------- } 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, } } Okay, I'm a biologist and have absolutely no idea of how to go about } setting } up a Philips CM-10 for dark field imaging. Now I have a user who needs it } and we need some help. } } I would appreciate some suggestions for basic settings to get us started } such as aperture sizes, spot size, tilt angles, etc. } } Thanks in advance, } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www.agriculture.purdue.edu/microscopy } } } ==============================Original } Headers============================== } 6, 21 -- From dsherman-at-purdue.edu Mon May 22 22:07:26 2006 } 6, 21 -- Received: from 1061exfe01.adpc.purdue.edu } (1061exfe01.adpc.purdue.edu [128.210.63.221]) } 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4N37QYM008244 } 6, 21 -- for {microscopy-at-microscopy.com} ; Mon, 22 May 2006 22:07:26 -0500 } 6, 21 -- Received: from exchange.purdue.edu ([128.210.63.234]) by } 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 6, 21 -- Mon, 22 May 2006 23:07:39 -0400 } 6, 21 -- Received: from 12.208.56.23 ([12.208.56.23]) by EXCH02.purdue.lcl } ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu } ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; } 6, 21 -- Tue, 23 May 2006 03:07:27 +0000 } 6, 21 -- User-Agent: Microsoft-Entourage/11.2.3.060209 } 6, 21 -- Date: Mon, 22 May 2006 23:07:26 -0400 } 6, 21 -- Subject: Dark field TEM } 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} } 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 6, 21 -- Message-ID: {C097F4AE.2A94%dsherman-at-purdue.edu} } 6, 21 -- Thread-Topic: Dark field TEM } 6, 21 -- Thread-Index: AcZ+FgS1Qwy8kuoJEdqGcAAKlcoUxg== } 6, 21 -- Mime-version: 1.0 } 6, 21 -- Content-type: text/plain; } 6, 21 -- charset="US-ASCII" } 6, 21 -- Content-transfer-encoding: 7bit } 6, 21 -- X-OriginalArrivalTime: 23 May 2006 03:07:39.0844 (UTC) } FILETIME=[0CF59840:01C67E16] } ==============================End of - } Headers============================== }
==============================Original Headers============================== 8, 25 -- From benada-at-biomed.cas.cz Tue May 23 02:03:07 2006 8, 25 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 8, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4N7359d023367 8, 25 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 02:03:06 -0500 8, 25 -- Received: from mail2.biomed.cas.cz (localhost [127.0.0.1]) 8, 25 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k4N71nfi016374; 8, 25 -- Tue, 23 May 2006 09:01:49 +0200 (CEST) 8, 25 -- Received: from 147.231.44.104 8, 25 -- (SquirrelMail authenticated user benada) 8, 25 -- by mail2.biomed.cas.cz with HTTP; 8, 25 -- Tue, 23 May 2006 09:01:49 +0200 (CEST) 8, 25 -- Message-ID: {1113.147.231.44.104.1148367709.squirrel-at-mail2.biomed.cas.cz} 8, 25 -- In-Reply-To: {200605230310.k4N3ACC4013129-at-ns.microscopy.com} 8, 25 -- References: {200605230310.k4N3ACC4013129-at-ns.microscopy.com} 8, 25 -- Date: Tue, 23 May 2006 09:01:49 +0200 (CEST) 8, 25 -- Subject: Re: [Microscopy] Dark field TEM 8, 25 -- From: =?iso-8859-2?Q?Old=F8ich_Benada?= {benada-at-biomed.cas.cz} 8, 25 -- To: dsherman-at-purdue.edu 8, 25 -- Cc: microscopy-at-microscopy.com 8, 25 -- User-Agent: SquirrelMail/1.4.6 8, 25 -- MIME-Version: 1.0 8, 25 -- Content-Type: text/plain;charset=iso-8859-2 8, 25 -- Content-Transfer-Encoding: 8bit 8, 25 -- X-Priority: 3 (Normal) 8, 25 -- Importance: Normal ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jeger-at-bgu.ac.il) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, May 23, 2006 at 05:05:26 ---------------------------------------------------------------------------
Email: jeger-at-bgu.ac.il Name: Rina Jeger
Organization: Ben Gurion University of the negev
Education: Graduate College
Location: City, State, Country
Question: We have a problem with blocks damaging the diamond knife. Cell cultures grown in flasks and processed without any glass cause us much damage. We don't use molecular sieves. We use Araldite and just the blocks of cells from the flasks cause the damage. Please help!
We had that problem and it turned out to be the microtome rather than the blocks. If bearings are worn out or advance motors are not working properly you can get enough instability in the cutting stroke to cause fine damage to the knife.
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: {jeger-at-bgu.ac.il} } Reply-To: {jeger-at-bgu.ac.il} } Date: Tue, 23 May 2006 08:15:46 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] AskAMicroscopist: Help, blocks damaging the diamond } knife } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (jeger-at-bgu.ac.il) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Tuesday, May 23, 2006 at 05:05:26 } --------------------------------------------------------------------------- } } Email: jeger-at-bgu.ac.il } Name: Rina Jeger } } Organization: Ben Gurion University of the negev } } Education: Graduate College } } Location: City, State, Country } } Question: We have a problem with blocks damaging the diamond knife. Cell } cultures grown in flasks and processed without any glass cause us much damage. } We don't use molecular sieves. We use Araldite and just the blocks of cells } from the flasks cause the damage. Please help! } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Tue May 23 08:12:22 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 } k4NDCKWF010300 } 7, 12 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 08:12:22 -0500 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) } 7, 12 -- Message-Id: {p06110400c098ba242815-at-[206.69.208.22]} } 7, 12 -- Date: Tue, 23 May 2006 08:12:19 -0500 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: jeger-at-bgu.ac.il (by way of Ask-A-Microscopist) } 7, 12 -- Subject: AskAMicroscopist: Help, blocks damaging the diamond knife } 7, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
==============================Original Headers============================== 7, 24 -- From dsherman-at-purdue.edu Tue May 23 08:43:36 2006 7, 24 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 7, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4NDhaB9021230 7, 24 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 08:43:36 -0500 7, 24 -- Received: from exchange.purdue.edu ([128.210.63.234]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 24 -- Tue, 23 May 2006 09:43:42 -0400 7, 24 -- 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 ; 7, 24 -- Tue, 23 May 2006 13:43:34 +0000 7, 24 -- User-Agent: Microsoft-Entourage/11.2.3.060209 7, 24 -- Date: Tue, 23 May 2006 09:43:33 -0400 7, 24 -- Subject: Re: [Microscopy] AskAMicroscopist: Help, blocks damaging the 7, 24 -- diamond knife 7, 24 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 24 -- To: {jeger-at-bgu.ac.il} , "message to: MSA list" {microscopy-at-microscopy.com} 7, 24 -- Message-ID: {C09889C5.10B7E%dsherman-at-purdue.edu} 7, 24 -- Thread-Topic: [Microscopy] AskAMicroscopist: Help, blocks damaging the 7, 24 -- diamond knife 7, 24 -- Thread-Index: AcZ+buIDIMhVhOpiEdq7bQARJN08Mg== 7, 24 -- In-Reply-To: {200605231315.k4NDFktC014502-at-ns.microscopy.com} 7, 24 -- Mime-version: 1.0 7, 24 -- Content-type: text/plain; 7, 24 -- charset="US-ASCII" 7, 24 -- Content-transfer-encoding: 7bit 7, 24 -- X-OriginalArrivalTime: 23 May 2006 13:43:42.0866 (UTC) FILETIME=[E7E49F20:01C67E6E] ==============================End of - Headers==============================
Hi, I always request that tissue culture flasks/dishes are rinsed with a sterile PBS prior to growing any cell lines to remove any debris that is in there. I had problems like that and I rinsed the flask, spun the content and checked under light microscope. There was plenty of debris, probably of plastic origin. It might ease the problem but it will not eliminate it 100%. If it is really bad I'd use a glass knife. Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Tue May 23 08:46:13 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4NDkDho026598 1, 22 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 08:46:13 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1FiXDA-0004yH-03 1, 22 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 10:46:12 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 23 May 06 10:46:12 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 23 May 06 10:45:27 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Tue, 23 May 2006 10:42:44 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: Help, blocks damaging the diamond knife 1, 22 -- Message-ID: {4472E723.20638.BBF91-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
I had this problem. The problem diminished when I switched from smashing osmium ampoules to dissolve the crystals to buying made up solutions.
Dave
-----Original Message----- X-from: jeger-at-bgu.ac.il [mailto:jeger-at-bgu.ac.il] Sent: 23 May 2006 14:14 To: David Patton
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jeger-at-bgu.ac.il) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, May 23, 2006 at 05:05:26 ------------------------------------------------------------------------ ---
Email: jeger-at-bgu.ac.il Name: Rina Jeger
Organization: Ben Gurion University of the negev
Education: Graduate College
Location: City, State, Country
Question: We have a problem with blocks damaging the diamond knife. Cell cultures grown in flasks and processed without any glass cause us much damage. We don't use molecular sieves. We use Araldite and just the blocks of cells from the flasks cause the damage. Please help!
A simple trick to eliminate smashing osmium vials is to dip the sealed ampoule into liquid nitrogen. This releases the osmium crystals from the glass walls. Then simply break the vial using an ampoule cracker (available through EM supply houses) and pour the osmium crystals into your bottle containing the water.
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: {David.Patton-at-uwe.ac.uk} } Reply-To: {David.Patton-at-uwe.ac.uk} } Date: Tue, 23 May 2006 09:10:02 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] RE: AskAMicroscopist: Help, } } } } } ---------------------------------------------------------------------------- } 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 had this problem. The problem diminished when I switched from } smashing osmium ampoules to dissolve the crystals to buying made up } solutions. } } Dave } } } } -----Original Message----- } X-from: jeger-at-bgu.ac.il [mailto:jeger-at-bgu.ac.il] } Sent: 23 May 2006 14:14 } To: David Patton } Subject: [Microscopy] AskAMicroscopist: Help, blocks damaging the } diamond knife } } } } } ------------------------------------------------------------------------ } ---- } 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 } ------------------------------------------------------------------------ } ---- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (jeger-at-bgu.ac.il) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Tuesday, May 23, 2006 at 05:05:26 } ------------------------------------------------------------------------ } --- } } Email: jeger-at-bgu.ac.il } Name: Rina Jeger } } Organization: Ben Gurion University of the negev } } Education: Graduate College } } Location: City, State, Country } } Question: We have a problem with blocks damaging the diamond knife. Cell } cultures grown in flasks and processed without any glass cause us much } damage. We don't use molecular sieves. We use Araldite and just the } blocks of cells from the flasks cause the damage. Please help! } } ------------------------------------------------------------------------ } --- } } ==============================Original } Headers============================== } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Tue May 23 08:12:22 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 k4NDCKWF010300 } 7, 12 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 } 08:12:22 -0500 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) } 7, 12 -- Message-Id: {p06110400c098ba242815-at-[206.69.208.22]} } 7, 12 -- Date: Tue, 23 May 2006 08:12:19 -0500 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: jeger-at-bgu.ac.il (by way of Ask-A-Microscopist) } 7, 12 -- Subject: AskAMicroscopist: Help, blocks damaging the diamond } knife } 7, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== } } } This incoming email to UWE has been independently scanned for viruses } and any virus detected has been removed using McAfee anti-virus software } } } } This email has been independently scanned for viruses and any virus software } has been removed using McAfee anti-virus software } } } ==============================Original Headers============================== } 24, 34 -- From David.Patton-at-uwe.ac.uk Tue May 23 09:07:12 2006 } 24, 34 -- Received: from mailapp01.uwe.ac.uk (mailapp01.uwe.ac.uk } [164.11.132.61]) } 24, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k4NE7Ap9009140 } 24, 34 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 09:07:11 -0500 } 24, 34 -- Received: from (164.11.132.60) by mailapp01.uwe.ac.uk via smtp } 24, 34 -- id 3bf4_d8fee282_ea65_11da_9721_0002b3c946e4; } 24, 34 -- Tue, 23 May 2006 15:10:11 +0100 } 24, 34 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk } 24, 34 -- (egen-uwe01.campus.ads.uwe.ac.uk [164.11.249.121]) } 24, 34 -- by mta01.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built } Jun 24 } 24, 34 -- 2005)) with ESMTP id {0IZQ0031O1VVKM-at-mta01.uwe.ac.uk} for } 24, 34 -- microscopy-at-microscopy.com; Tue, 23 May 2006 15:07:07 +0100 (BST) } 24, 34 -- Date: Tue, 23 May 2006 15:07:07 +0100 } 24, 34 -- From: David Patton {David.Patton-at-uwe.ac.uk} } 24, 34 -- Subject: RE: [Microscopy] AskAMicroscopist: Help, } 24, 34 -- blocks damaging the diamond knife } 24, 34 -- To: microscopy-at-microscopy.com } 24, 34 -- Message-id: {F247F674896BE243AD8263C5280E2BDB024A649F-at-egen-uwe01} } 24, 34 -- MIME-version: 1.0 } 24, 34 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.6944.0 } 24, 34 -- Content-type: text/plain; charset=us-ascii } 24, 34 -- Content-class: urn:content-classes:message } 24, 34 -- Thread-topic: [Microscopy] AskAMicroscopist: Help, } 24, 34 -- blocks damaging the diamond knife } 24, 34 -- Thread-index: AcZ+as/wtnGa9HTrS4+NkX8JBfbkTQABwBcA } 24, 34 -- X-MS-Has-Attach: } 24, 34 -- X-MS-TNEF-Correlator: } 24, 34 -- X-NAI-Spam-Score: -1.2 } 24, 34 -- X-NAI-Spam-Rules: 1 Rules triggered } 24, 34 -- BAYES_01=-1.2 } 24, 34 -- X-NAIMIME-Disclaimer: 1 } 24, 34 -- X-NAIMIME-Modified: 1 } 24, 34 -- Content-Transfer-Encoding: 8bit } 24, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k4NE7Ap9009140 } ==============================End of - Headers==============================
==============================Original Headers============================== 8, 23 -- From dsherman-at-purdue.edu Tue May 23 09:24:30 2006 8, 23 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4NEOTLs019419 8, 23 -- for {microscopy-at-microscopy.com} ; Tue, 23 May 2006 09:24:30 -0500 8, 23 -- Received: from exchange.purdue.edu ([128.210.63.234]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 8, 23 -- Tue, 23 May 2006 10:24:29 -0400 8, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 8, 23 -- Tue, 23 May 2006 14:24:28 +0000 8, 23 -- User-Agent: Microsoft-Entourage/11.2.3.060209 8, 23 -- Date: Tue, 23 May 2006 10:24:28 -0400 8, 23 -- Subject: Re: [Microscopy] RE: AskAMicroscopist: Help, 8, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 8, 23 -- To: {David.Patton-at-uwe.ac.uk} , 8, 23 -- "message to: MSA list" {microscopy-at-microscopy.com} 8, 23 -- Message-ID: {C098935C.10B8F%dsherman-at-purdue.edu} 8, 23 -- Thread-Topic: [Microscopy] RE: AskAMicroscopist: Help, 8, 23 -- Thread-Index: AcZ+dJlO16nmvupnEdq7bQARJN08Mg== 8, 23 -- In-Reply-To: {200605231410.k4NEA2OE013293-at-ns.microscopy.com} 8, 23 -- Mime-version: 1.0 8, 23 -- Content-type: text/plain; 8, 23 -- charset="US-ASCII" 8, 23 -- Content-transfer-encoding: 7bit 8, 23 -- X-OriginalArrivalTime: 23 May 2006 14:24:29.0124 (UTC) FILETIME=[99F9F840:01C67E74] ==============================End of - Headers==============================
On May 22, 2006, at 8:07 PM, dsherman-at-purdue.edu wrote:
} Okay, I'm a biologist and have absolutely no idea of how to go about } setting } up a Philips CM-10 for dark field imaging. Now I have a user who } needs it } and we need some help. } } I would appreciate some suggestions for basic settings to get us } started } such as aperture sizes, spot size, tilt angles, etc. } Dear Debby, I have only used dark field on our Tecnai scopes, which have a push-button to select dark field, but the process is likely to be the same on the CM10. The settings used will depend on what the user is trying to see. To get an idea, take a typical area of the specimen--or a positive control--and look in diffraction mode with no objective aperture to see where the signal you want is. When dark field is activated, the diffraction pattern shifts by an amount that varies with the tilt angle, so if you increase the tilt from zero, there will be a value that puts the signal you want to see at the position that the unscattered beam occupies for zero tilt, and this will be the tilt angle you want. If you now turn off dark field, insert and center an objective aperture, and turn on dark field, you will see what part of the diffracted beam will constitute the dark field image. The smaller the aperture, the more selective it will be, but the smaller the signal that will get through. In any case, you want an aperture that blocks the unscattered beam, and its maximum size will depend on the dark field tilt angle you chose. If you have crystalline material, the aperture will select signal from only a subset of possible orientations, which could either be what you want or just a nuisance. If the latter, dynamical dark field, AKA hollow-cone illumination, will solve it, but on the Tecnai scopes, it is only available with STEM. Spot size should be selected to give you sufficient signal, and other than that it will have little effect on the DF image unless you are using an extremely small objective aperture, in which case, a smaller spot size number will smear the diffraction rings or spots and lessen the amount and selectivity of the signal. For taking DF images of terbium, a 1 deg tilt and 70 um objective aperture allowed me to see the terbium as bright areas with faint gray corresponding to places where lower-Z material was and very few counts corresponding to blank areas of the grid. Good luck. 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 May 23 12:33:44 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 k4NHXih3001067 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 23 May 2006 12:33:44 -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 7A90535194 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 23 May 2006 10:33:43 -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 id 918E6352F7 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 23 May 2006 10:32:39 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200605230307.k4N37ZPp008420-at-ns.microscopy.com} 4, 22 -- References: {200605230307.k4N37ZPp008420-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {6f29c4ec2f0ff590d96623cd69b25b92-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Dark field TEM 4, 22 -- Date: Tue, 23 May 2006 10:43:38 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Hello, I'm seeking reccomendations for a small CO2 incubator to use in an imaging facility to hold samples waiting for their turn on the microscope. This period could be as long as several hours. Any suggestions for 1 cubic foot (0.028 m^3) to 1.4 cubic foot (0.039 m^3) incubator? Have air jacketed been sufficient or does anyone wish they'd purchased a water jacketed incubator instead?
thanks, glen
Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
==============================Original Headers============================== 9, 24 -- From glenmac-at-u.washington.edu Tue May 23 13:47:34 2006 9, 24 -- Received: from mxout2.cac.washington.edu (mxout2.cac.washington.edu [140.142.33.4]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4NIlXW6013467 9, 24 -- for {Microscopy-at-sparc5.microscopy.com} ; Tue, 23 May 2006 13:47:34 -0500 9, 24 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 9, 24 -- by mxout2.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k4NIlW5H008609 9, 24 -- for {Microscopy-at-sparc5.microscopy.com} ; Tue, 23 May 2006 11:47:33 -0700 9, 24 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 9, 24 -- (authenticated authid=glenmac) 9, 24 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k4NIlUBH023421 9, 24 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 9, 24 -- for {Microscopy-at-sparc5.microscopy.com} ; Tue, 23 May 2006 11:47:32 -0700 9, 24 -- Mime-Version: 1.0 (Apple Message framework v750) 9, 24 -- In-Reply-To: {l03130300ba5f4ec544de-at-[152.83.167.45]} 9, 24 -- References: {l03130300ba5f4ec544de-at-[152.83.167.45]} 9, 24 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 24 -- Message-Id: {B22D3EB6-1604-4AF3-BDE7-49AD22C6D6F6-at-u.washington.edu} 9, 24 -- Content-Transfer-Encoding: 7bit 9, 24 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 9, 24 -- Subject: incubator for holding samples 9, 24 -- Date: Tue, 23 May 2006 11:47:28 -0700 9, 24 -- To: Microscopy List Server {Microscopy-at-ns.microscopy.com} 9, 24 -- X-Mailer: Apple Mail (2.750) 9, 24 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 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' ==============================End of - Headers==============================
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Have you got the holder connected to the power supply, temperature sensor, etc?
It's possible the outer part of central furnace is grounded via the the electrical connections - if you don't have it connected up, it may be charging up and deflecting the beam. Even if you have it plugged in, if the holder hasn't been used for several years, there may be a broken connection.
PLEASE NOTE - Agressive SPAM filtering is employed. If you don't get a reply to e-mails, try again, avoiding anything in the subject or body which might trigger filtering. If you are in the address book, you should get through. If you aren't, then there's a chance your e-mail will never be seen.
==============================Original Headers============================== 5, 17 -- From larry-at-cymru.freewire.co.uk Tue May 23 14:59:58 2006 5, 17 -- Received: from get.freewire.net ([86.54.106.202]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4NJxwNp024749 5, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 23 May 2006 14:59:58 -0500 5, 17 -- Received: from [217.154.251.138] (th6dc-217-154-251-138.dial.mistral.co.uk [217.154.251.138] (may be forged)) 5, 17 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k4NJxsAT028594; 5, 17 -- Tue, 23 May 2006 20:59:55 +0100 5, 17 -- Mime-Version: 1.0 5, 17 -- Message-Id: {p06210201c09918dae69b-at-[217.154.254.6]} 5, 17 -- In-Reply-To: {200605230026.k4N0QhrW001980-at-ns.microscopy.com} 5, 17 -- References: {200605230026.k4N0QhrW001980-at-ns.microscopy.com} 5, 17 -- Date: Tue, 23 May 2006 20:58:54 +0100 5, 17 -- To: roderi_co-at-yahoo.com, Microscopy-at-MSA.Microscopy.Com 5, 17 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 17 -- Subject: Re: [Microscopy] viaWWW: TEM - Problem with heating specimen 5, 17 -- holder 5, 17 -- 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/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both istillma-at-bidmc.harvard.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: istillma-at-bidmc.harvard.edu Name: Isaac E Stillman
Organization: BIDMC/Harvard Medical School
Title-Subject: [Filtered] Advice
Question: Hi-
I am a renal pathologist who is purchasing a new instrument for our diagnostic EM unit. The choice has come down to FEI (Phillips) Morgagni vs JEOL JEM-1011. Of course, we would be adding a 2K digital camera. I would be most grateful to hear your thoughts on this choice, off list if you prefer.
Thanks! Isaac E Stillman MD Dept. of Pathology - BIDMC/HMS
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both huisheng.jiao-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: huisheng.jiao-at-gmail.com Name: Ding
Organization: The University of Birmingham
Title-Subject: [Filtered] Suva 124 (R124)?
Question: Does anybody have experience in Suva 124 (R124) as cryogen for rapid freezing?
In the late 80's early 90's M. Parthasarathy, Cornell University, & Theo Muller, BAL-TEC AG published a paper examining using SUVA 124 for cryofixation.
Best,
Al Coritz, Technical Engineer Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- X-from: {huisheng.jiao-at-gmail.com} To: {Sampleprep-at-earthlink.net} Sent: Wednesday, May 24, 2006 8:43 AM
Dear Al, dear Huisheng,
I'v found in the MSA listserver-Archives cf: http://www.microscopy.com/cgi-bin/ReadPrintEmailHTML.pl?filename=9807.txt
a short thread on the possible use of SUVA, in which perhaps also the Article mentioned below has been referenced: It says (for the whole thread use "FREON" or "replacement" as a search phrase): } } } } } } } } } } } } } } } } } } } } { { { { { { { { { { { { { { { { { { { { { { { { Re: Freon replacements for cryo
Try Muller T, Moser S, Vogt M, Daugherty C & Parasarathy MV (1993). Optimisation and application of jet freezing. Scanning Microscopy 7, 1295-1310.
Using HCFC 124 (SUVA 124-CHClFCF3) with thin titanium supports, cooling rates were obtained similar to those from propane and standard copper supports. This was suggested (I hesitate to plug this !!) in Ryan KP (1992) Cryofixation of tissues for electron microsocpy: a review of plunge cooling methods. Scanning Microsc. 6, 715-743. See next-to-last page , p. 742 in Discussion with Reviewers section..
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In the late 80's early 90's M. Parthasarathy, Cornell University, & Theo Muller, BAL-TEC AG published a paper examining using SUVA 124 for cryofixation.
Best,
Al Coritz, Technical Engineer Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- X-from: {huisheng.jiao-at-gmail.com} To: {Sampleprep-at-earthlink.net} Sent: Wednesday, May 24, 2006 8:43 AM
Micromavens,
Has anyone on the list had recent experience shipping small flowers fixed for EM? Or bringing such samples along as baggage on an airplane? I've done similar things in the past, but not recently, and times and regulations have changed. What's allowed? What's not? This is specifically for flowers (small ones) and flights or courier shipments originating and ending in the USA. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Wed May 24 14:06:53 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4OJ6rJS007173 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 24 May 2006 14:06:53 -0500 3, 20 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k4OJgS4p025733 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 24 May 2006 15:42:32 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 3, 20 -- Wed, 24 May 2006 15:06:48 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f0623090fc09a5e70a809-at-[141.209.160.132]} 3, 20 -- Date: Wed, 24 May 2006 15:06:48 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: shipping flowers 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 24 May 2006 19:06:48.0792 (UTC) FILETIME=[3537ED80:01C67F65] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -4 () L_EXCH_MF 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
I've been asked to seek opinions from folks who have used 2 or more of the currently available knife-makers: The GMK triangular knife maker sold by Energy Beam Sciences, and the two "balanced break" knife-makers, the Leica EM KMR 2 and the RMC GKM (an unfortunately confusing model designation). What do folks think of them, and how easily can novices be taught to consistently make good knives usable for thin sectioning? Of these, I've only used the GMK, formally known as the Sorvall brick. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 2, 20 -- From oshel1pe-at-cmich.edu Wed May 24 14:11:48 2006 2, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4OJBl5O012814 2, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 24 May 2006 14:11:48 -0500 2, 20 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 2, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k4OJlR4l026099 2, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 24 May 2006 15:47:27 -0400 2, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 2, 20 -- Wed, 24 May 2006 15:11:45 -0400 2, 20 -- Mime-Version: 1.0 2, 20 -- Message-Id: {f06230910c09a5f52dd0d-at-[141.209.160.132]} 2, 20 -- Date: Wed, 24 May 2006 15:11:46 -0400 2, 20 -- To: Microscopy-at-microscopy.com 2, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 2, 20 -- Subject: glass knife makers 2, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 2, 20 -- X-OriginalArrivalTime: 24 May 2006 19:11:46.0183 (UTC) FILETIME=[E67A3170:01C67F65] 2, 20 -- X-CanItPRO-Stream: default 2, 20 -- X-Spam-Score: -4 () L_EXCH_MF 2, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
We're plannig to build a sterotaxic instrument for animal brain studies. It must be adaptable for mouse, rat and rabbit. We're looking for plans, drawings etc. anything beneficial. Does anybody know this kind of materials reachable on internet or another source?
Thanks in advance...
Dr. Necat Yilmaz Mersin University School of Medicine Histology and Embryology Dept. Mersin/TURKEY
==============================Original Headers============================== 4, 31 -- From nyilmaz-at-mersin.edu.tr Thu May 25 02:07:31 2006 4, 31 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 4, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4P77Sha015028 4, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 25 May 2006 02:07:31 -0500 4, 31 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 4, 31 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 3C5D7480A7 4, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 25 May 2006 10:07:20 +0300 (EEST) 4, 31 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 4, 31 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 4, 31 -- with ESMTP id 02205-49 for {Microscopy-at-microscopy.com} ; 4, 31 -- Thu, 25 May 2006 10:07:06 +0300 (EEST) 4, 31 -- Received: from NEJAT1 (unknown [193.255.128.132]) 4, 31 -- by mail.mersin.edu.tr (Postfix) with SMTP id 9CCDA48081 4, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 25 May 2006 10:07:06 +0300 (EEST) 4, 31 -- Message-ID: {002501c67fc9$d53656a0$4d01a8c0-at-NEJAT1} 4, 31 -- From: "Nejat" {nyilmaz-at-mersin.edu.tr} 4, 31 -- To: "EM-Mail Group" {Microscopy-at-microscopy.com} 4, 31 -- Subject: Building a Stereotaxic Instrument 4, 31 -- Date: Thu, 25 May 2006 09:35:08 +0300 4, 31 -- MIME-Version: 1.0 4, 31 -- Content-Type: text/plain; 4, 31 -- format=flowed; 4, 31 -- charset="windows-1254"; 4, 31 -- reply-type=original 4, 31 -- Content-Transfer-Encoding: 7bit 4, 31 -- X-Priority: 3 4, 31 -- X-MSMail-Priority: Normal 4, 31 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 4, 31 -- Disposition-Notification-To: "Nejat" {nyilmaz-at-mersin.edu.tr} 4, 31 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 4, 31 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr ==============================End of - Headers==============================
Dear friends I am intersted in the following pdf reprint or xerox if possible (fax is in the signature). As well I use this occasion to remind that we have two schools in Genoa, namely: Fluorescence 19-22 June; Confocal and Multiphoton microscopy 4-7 July. SInce the number of partecipants is limited, please have a check at www.lambs.it or send an e.-mail to me for reservations. All my best Alby
paper ref Squeezing in two photon absorption from a strong coherent beam G. S. Agarwal Optics Communications Pages 190-192, Volume 62, Issue 3, 1 May, 1987.
--------------------------------------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001) --------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ----------------------------------------------
==============================Original Headers============================== 8, 18 -- From diaspro-at-fisica.unige.it Thu May 25 02:39:43 2006 8, 18 -- Received: from phobos.ge.infm.it (phobos.ge.infm.it [193.205.153.2]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4P7dg2V025623; 8, 18 -- Thu, 25 May 2006 02:39:43 -0500 8, 18 -- Received: from [193.205.153.216] (albypc.ge.infm.it [193.205.153.216]) 8, 18 -- by phobos.ge.infm.it (8.11.6/8.11.0) with ESMTP id k4P7coh25601; 8, 18 -- Thu, 25 May 2006 09:38:50 +0200 8, 18 -- Mime-Version: 1.0 (Apple Message framework v750) 8, 18 -- Content-Transfer-Encoding: 7bit 8, 18 -- Message-Id: {AB7F2E4C-2044-4652-B9A4-0A154009B743-at-fisica.unige.it} 8, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 18 -- To: Confocal List Microscopy {CONFOCAL-at-LISTSERV.BUFFALO.EDU} , 8, 18 -- mplsm-users-at-yahoogroups.com, microscopy-at-microscopy.com, 8, 18 -- MicroscopyListSpamFilter-at-microscopy.com 8, 18 -- From: diaspro {diaspro-at-fisica.unige.it} 8, 18 -- Subject: reprint help alby 8, 18 -- Date: Thu, 25 May 2006 09:34:22 +0200 8, 18 -- X-Mailer: Apple Mail (2.750) ==============================End of - Headers==============================
We have been using a flatbed scanner for documenting rock core before processing it for other analytical techniques. Now, we want to explore the technique a bit further, possibly towards applied image analysis, but need one last ingredient for better quality scans. We want to use some akin to immersion oil for the interface between rock and flatbed, but want to find something inexpensive, non-toxic and easy to clean. Any thoughts?
Hi, Our lab has been using Leica EM KMR 2 for a few years now. So far we have no problem with the machine. It is reliable and makes consistently good knives. Dorota
==============================Original Headers============================== 1, 24 -- From wadowska-at-avcn1.novell.upei.ca Thu May 25 06:31:43 2006 1, 24 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4PBVhX9019417 1, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 25 May 2006 06:31:43 -0500 1, 24 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 24 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 24 -- id 1FjE47-0001zh-01 1, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 25 May 2006 08:31:43 -0300 1, 24 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 24 -- 25 May 06 08:31:43 -0300 1, 24 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 25 May 06 08:30:44 -0300 1, 24 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 24 -- Organization: University of P.E.I. 1, 24 -- To: microscopy-at-msa.microscopy.com 1, 24 -- Date: Thu, 25 May 2006 08:27:27 -0400 1, 24 -- MIME-Version: 1.0 1, 24 -- Content-type: text/plain; charset=US-ASCII 1, 24 -- Content-transfer-encoding: 7BIT 1, 24 -- Subject: [Microscopy] glass knife makers 1, 24 -- Message-ID: {44756A6F.16193.11BB3E-at-localhost} 1, 24 -- X-Confirm-Reading-To: "Dorota Wadowska" {wadowska-at-acad1.cs.upei.ca} 1, 24 -- X-pmrqc: 1 1, 24 -- Priority: normal 1, 24 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
If you feel you would like to use immersion oil, why don't you - non fluorescent immersion oil is pretty cheap from the likes of Zeiss and Cargille, and isn't particularly toxic (the anti-fluorescent variety is a little more toxic). As long as you keep the oil in the centre of the glass platen and don't shut the platen lid down, it's not going anywhere. In fact you can get quite good scans of things like fruit, seeds, leaves, calculators, small toys etc.. using film scanning flatbeds in reflective mode at high resolutions (certainly better than the likes of that from an $80 QX-5 toy microscope). You may need to use Silverfast.com twain software to overcome the scanners reluctance to go above 1,200 dpi with a reflective A4 scan though. Also many scanners are fine focussed a little above the glass platen, as it is assumed you are using film in holders. The superb Epson V750 pro flatbed scanner even has optional little legs on the film holders to help with this. I'm pretty sure that the scanner optics aren't optimised for oil immersion though, it being an air interface on the underneath of the scanner. You won't get good scan results with a cheap LiDE type scanner (no depth of field), you really need a film scanner type flatbed.
You can easily remove immersion oil with 50:50 alcohol water and tissues, in fact I use 50:50 propanol and water regularly on my Canon 9950F and Epson 4990 photo flatbeds (naturally avoiding the platen edges when spraying). This mixture also cleans all our imaging workstation's VDU CRT screens (removing all finger marks). Also try 100% ether for stubborn stains. Fingermarks are a similar oily mess to immersion oil, but it's only non removable scratches from sharp rocks that are the real pain (try using the sample on glass cover slips or slides perhaps ?). You can try using things like thin plastic strips (or thin card with film) to scoop under the sample to remove it, without touching the platen glass.
Keith
--------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: michael-at-Shaffer.net [mailto:michael-at-Shaffer.net] Sent: 25 May 2006 11:57 To: keith.morris-at-ucl.ac.uk
We have been using a flatbed scanner for documenting rock core before processing it for other analytical techniques. Now, we want to explore the technique a bit further, possibly towards applied image analysis, but need one last ingredient for better quality scans. We want to use some akin to immersion oil for the interface between rock and flatbed, but want to find something inexpensive, non-toxic and easy to clean. Any thoughts?
Glycerin comes to mind as being cheap, non-toxic, easier to clean, clear, refractive index ~1.47 and just all around more "friendly" than a traditional refractive index liquid. I've never used it, but it comes to mind as something I have seen mentioned as usable for immersion media. There are lots of household liquids that you might also consider, vegetable oil, olive oil, and mineral oil come to mind.
Good Luck, Bryan Bandli
michael-at-Shaffer.net wrote:
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You could try water as well. Glycerol can be diluted in it. We regularly use water or glycerol immersion and it works great with special water/glycerol immersion objectives.
On a scanner in reflected mode, oil can really shine sometimes though (particularly at oil/air interfaces). Some vegetable oils such as corn oil also evaporate to leave a very hard resin residue that can't easily be removed (naturally immersion oil doesn't) - so the glass must be cleaned well afterwards. If your sample is huge (6 inches or more) I would have thought any cheap clear mineral oil would be ideal (there are plenty to chose from, even baby oil). On a flat polished sample you shouldn't need much oil.
If a sample is very very thin (we used annular saws with our hard human bone samples and ground the surfaces flat - so I'm thinking along those lines) - and you can get some light through it (or you are looking at something like loose soil particles), running the scanner in transmission film scan mode might work. You have more than half a centimetre or so between the platen glass and the film scanner lid with an Epson 4990 photo, so you can get a thin sheet of glass (e.g. slide thickness) and a thin sample in there. Samples in oil appear to scan better in this film scan mode than in reflected mode, particularly at air/oil surfaces.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: bbandli-at-mvainc.com [mailto:bbandli-at-mvainc.com] Sent: 25 May 2006 14:08 To: keith.morris-at-ucl.ac.uk
Michael,
Glycerin comes to mind as being cheap, non-toxic, easier to clean, clear, refractive index ~1.47 and just all around more "friendly" than a traditional refractive index liquid. I've never used it, but it comes to mind as something I have seen mentioned as usable for immersion media. There are lots of household liquids that you might also consider, vegetable oil, olive oil, and mineral oil come to mind.
Good Luck, Bryan Bandli
michael-at-Shaffer.net wrote:
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Normal mineral oil from the drug store has RI 1.51. I would try it on a glass surface other than the scanner bed first, to make sure that you don't get moire patterns. Also, try various cleaning methods. I think that wiping it dry first then using either Windex or rubbing alcohol should do the trick.
Good hunting! 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.
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At 05:55 AM 5/25/2006, michael-at-Shaffer.net wrote:
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Hi Mike, I have been exploring and having success using a flatbed scanner for particle analysis, but actually started out in this endeavor by scanning rocks and rock cores. We now use an Epson XL 1640 scanner, which has a focus function, essentially allowing us to get right on the surface of the samples. However, a pretty flat cut is needed to get overall excellent quality. I found that this works much better than greasing up the bed with oils, glycerin or other sticky stuff. We used a thin plastic sheet (same material as thin clear view-foils) to protect the glass from the rocks. As a side note, I too saw some strange artifacts with oils, which could (?) be due to "internal" reflections or interference patterns between the glass-plastic or micro-bubbles on or close the sample surface. My major concern was contaminating the samples with oils, especially when doing trace element work or isotope analysis for age dating purposes of the rock cores. Not that the oil itself would be the major culprit, but more the rock dust flying around everywhere in the cutting lab sticking to the greasy surfaces. Rock cores sometimes have oil on them from the drilling process and is usually washed off with sulfo. However, the porosity of the sample may contribute to deeper contamination, especially is if the sample shows cracks or micro-fractures. I had to consider this for archival purposes and potential future analysis on the same samples as new technologies emerge.
michael-at-Shaffer.net wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } We have been using a flatbed scanner for documenting rock core before } processing it for other analytical techniques. Now, we want to explore the } technique a bit further, possibly towards applied image analysis, but need } one last ingredient for better quality scans. We want to use some akin to } immersion oil for the interface between rock and flatbed, but want to find } something inexpensive, non-toxic and easy to clean. Any thoughts? } } TIA & 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 } } } } } ==============================Original Headers============================== } 7, 19 -- From Michael-at-Shaffer.net Thu May 25 05:52:02 2006 } 7, 19 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4PAq1qK008574 } 7, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 25 May 2006 05:52:01 -0500 } 7, 19 -- Received: (qmail 17426 invoked from network); 25 May 2006 10:52:00 -0000 } 7, 19 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) } 7, 19 -- by ws6-4.us4.outblaze.com with SMTP; 25 May 2006 10:52:00 -0000 } 7, 19 -- From: "michael shaffer" {michael-at-Shaffer.net} } 7, 19 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} } 7, 19 -- Subject: imaging rock core with flatbed scanner } 7, 19 -- Date: Thu, 25 May 2006 08:21:59 -0230 } 7, 19 -- Message-ID: {000f01c67fe9$40c556e0$8d829986-at-roamingwolf} } 7, 19 -- MIME-Version: 1.0 } 7, 19 -- Content-Type: text/plain; } 7, 19 -- charset="US-ASCII" } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- X-Mailer: Microsoft Office Outlook 11 } 7, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 7, 19 -- Thread-Index: AcZ/6T9zCvUQjGd1R8+CGbe9ZHz2pA== } ==============================End of - Headers============================== } }
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Email: mccaulak-at-wfu.edu Name: Anita McCauley
Organization: Wake Forest University
Title-Subject: [Filtered] procedure for quantitative fluorescence
Question: I am looking for advice for doing quantitative fluorescence microscopy. My focus is on the microscope, camera, and software end of it, rather than the sample prep. Last year, a question about quantitative fluorescence on this list resulted in a number of very helpful posts about things to do, like controlling camera settings and using fluorescence reference slides. However, what is not clear is how to combine all these things together into an appropriate procedure. If anybody could help with any of the following, I would greatly appreciate it:
1. I purchased a set of fluorescent reference slides but have been unable to obtain any documentation or instructions from the supplier for how to use them correctly for fluorescent imaging.
2. When comparing a signal between two samples, is it best to subtract the background from the signal or just compare the two signals?
3. To determine the signal, is it best to use irregular AOIs and get a mean or summed value or to use multiple line profiles, or something else?
4. If anyone would be willing to share a step-by-step protocol that they have developed to help users do image capture and analysis of fluorescence correctly, I would greatly appreciate it.
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Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers-Institute
Title-Subject: [Filtered] Lowicryl
Question: Good Afternoon,
I am looking for comment's in regards to embedding in Lowicryl HM-20 in the Leica AFS. I have done some reading, but have never attempted it before. We will be using HPF-AFSyeast samples at first. Fixative's will be 2% Gluteraldehyde/0.5% Uranyl Acetate in acetone (for morphology) and a 0.1% Gluteraldehyde/0.1% Uranyl acetate in acetone. My question's are:
1. What kind of formulation's for final resin concentration are people using?
2. I am afraid some of the sample's will fall through the holes in the specimen container's as you changes solutions. Are there any suggestion's (Besides not letting them be so tiny and digging them out with a wooden stick) to prevent that?
3. Most people are using the flat embedding molds?
4. Using -50 C UV polymerization for how long?
5. How many say stay away from Lowicryl HM-20 for another immuno-friendly resing?
Thanks in advance for your help. Will probably have more questions as I go through the process.
Rhonda Allen Stower's Institute Kansas City, MO 816-926-4305
Hello, I've been processing some small plant roots for scanning electron microscopy. I use Glutaraldehyde fixation followed by 0.8% KFerrocyanide and 1% OsO4 - otherwise standard fixation. I get great root preservation, but I've found that the root hairs all look very collapsed. I can send a picture if you like, but was wondering if anyone had experience in this area.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 2, 24 -- From gvrdolja-at-nature.berkeley.edu Thu May 25 18:14:50 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 k4PNEo4t017400 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 25 May 2006 18:14:50 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id AEC69C1E39 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 25 May 2006 16:14:49 -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 25387-03 for {microscopy-at-microscopy.com} ; 2, 24 -- Thu, 25 May 2006 16:14:37 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id 5E8A1C1E57; Thu, 25 May 2006 16:14:37 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 321C1C1E55 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 25 May 2006 16:14:36 -0700 (PDT) 2, 24 -- Date: Thu, 25 May 2006 16:14:36 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: microscopy-at-microscopy.com 2, 24 -- Subject: roots for EM 2, 24 -- Message-ID: {Pine.SOC.4.64.0605251612590.20397-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==============================
We are using the Leica EM KMR2 without problems. It is easy to learn and to use and is a reliable instrument.
Stephane
--- oshel1pe-at-cmich.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've been asked to seek opinions from folks who have } used 2 or more } of the currently available knife-makers: } The GMK triangular knife maker sold by Energy Beam } Sciences, and the } two "balanced break" knife-makers, the Leica EM KMR } 2 and the RMC GKM } (an unfortunately confusing model designation). } What do folks think of them, and how easily can } novices be taught to } consistently make good knives usable for thin } sectioning? } Of these, I've only used the GMK, formally known as } the Sorvall brick. } Thanks. } } Phil } -- } Philip Oshel } Microscopy Facility Supervisor } Department of Biology } Central Michigan University } 024C Brooks Hall } Mt. Pleasant, MI 48859 } (989) 774-3576 } } ==============================Original } Headers============================== } 2, 20 -- From oshel1pe-at-cmich.edu Wed May 24 14:11:48 } 2006 } 2, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu } [141.209.20.21]) } 2, 20 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k4OJBl5O012814 } 2, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 24 } May 2006 14:11:48 -0500 } 2, 20 -- Received: from egateb.central.cmich.local } ([141.209.15.85]) } 2, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with } ESMTP id k4OJlR4l026099 } 2, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 24 } May 2006 15:47:27 -0400 } 2, 20 -- Received: from [141.209.160.132] } ([141.209.160.132]) by egateb.central.cmich.local } with Microsoft SMTPSVC(5.0.2195.6713); } 2, 20 -- Wed, 24 May 2006 15:11:45 -0400 } 2, 20 -- Mime-Version: 1.0 } 2, 20 -- Message-Id: } {f06230910c09a5f52dd0d-at-[141.209.160.132]} } 2, 20 -- Date: Wed, 24 May 2006 15:11:46 -0400 } 2, 20 -- To: Microscopy-at-microscopy.com } 2, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} } 2, 20 -- Subject: glass knife makers } 2, 20 -- Content-Type: text/plain; } charset="us-ascii" ; format="flowed" } 2, 20 -- X-OriginalArrivalTime: 24 May 2006 } 19:11:46.0183 (UTC) FILETIME=[E67A3170:01C67F65] } 2, 20 -- X-CanItPRO-Stream: default } 2, 20 -- X-Spam-Score: -4 () L_EXCH_MF } 2, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . } com) on 141.209.20.21 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 8, 20 -- From nizets2-at-yahoo.com Fri May 26 02:40:06 2006 8, 20 -- Received: from web37409.mail.mud.yahoo.com (web37409.mail.mud.yahoo.com [209.191.87.62]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4Q7e5aY005571 8, 20 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 02:40:05 -0500 8, 20 -- Received: (qmail 70344 invoked by uid 60001); 26 May 2006 07:40:05 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=X4G697hdE+bmWEJLeyUOWho0vsDx+HVX5N7uvQiaoEhWsjB8t2wHo5ACGkTEITHks5ILmqDY87qACfUo5wkmeKDn4wFbuQkW51bOIwBRcTI9MONTWV3tHmyGHfTZAuvefrlzZ2g/1R/Rj5DFgckwsVLuarooj3eHvucGf/uCGds= ; 8, 20 -- Message-ID: {20060526074005.70342.qmail-at-web37409.mail.mud.yahoo.com} 8, 20 -- Received: from [80.122.101.102] by web37409.mail.mud.yahoo.com via HTTP; Fri, 26 May 2006 00:40:05 PDT 8, 20 -- Date: Fri, 26 May 2006 00:40:05 -0700 (PDT) 8, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 20 -- Subject: Re: [Microscopy] glass knife makers 8, 20 -- To: oshel1pe-at-cmich.edu 8, 20 -- Cc: microscopy-at-microscopy.com 8, 20 -- In-Reply-To: {200605241916.k4OJGIK9025364-at-ns.microscopy.com} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
My answer will deal with confocal microscopy: I use to insert my negative control and choose the lasers and detection settings so that I get no signal. Then I insert my samples and won't change the settings. To compare fluorescence intensities, I use to draw a profile line and compare the peaks (maximum intensities). I do it because I observe compact homogeneous bodies, but I suppose I would draw a ROI in the case I had to compare the total intensity of cell cytoplasm for example. It all depends on what and how you measure, there is no general rule I think, it must be adapted to be as "ethical" (yes again ;-)) as possible.
regards,
Stéphane
--- mccaulak-at-wfu.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the } Microscopy Listserver } using the WWW based Form at } http://www.microscopy.com/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a } Subscriber, so when replying } please copy both mccaulak-at-wfu.edu as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: mccaulak-at-wfu.edu } Name: Anita McCauley } } Organization: Wake Forest University } } Title-Subject: [Filtered] procedure for quantitative } fluorescence } } Question: I am looking for advice for doing } quantitative fluorescence microscopy. My focus is } on the microscope, camera, and software end of it, } rather than the sample prep. Last year, a question } about quantitative fluorescence on this list } resulted in a number of very helpful posts about } things to do, like controlling camera settings and } using fluorescence reference slides. However, what } is not clear is how to combine all these things } together into an appropriate procedure. If anybody } could help with any of the following, I would } greatly appreciate it: } } 1. I purchased a set of fluorescent reference slides } but have been unable to obtain any documentation or } instructions from the supplier for how to use them } correctly for fluorescent imaging. } } 2. When comparing a signal between two samples, is } it best to subtract the background from the signal } or just compare the two signals? } } 3. To determine the signal, is it best to use } irregular AOIs and get a mean or summed value or to } use multiple line profiles, or something else? } } 4. If anyone would be willing to share a } step-by-step protocol that they have developed to } help users do image capture and analysis of } fluorescence correctly, I would greatly appreciate } it. } } Thanks. } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 11, 12 -- From zaluzec-at-microscopy.com Thu May 25 } 17:51:32 2006 } 11, 12 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 11, 12 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k4PMpVYQ029216 } 11, 12 -- for {microscopy-at-microscopy.com} ; Thu, 25 } May 2006 17:51:32 -0500 } 11, 12 -- Mime-Version: 1.0 } 11, 12 -- X-Sender: (Unverified) } 11, 12 -- Message-Id: } {p06110401c09be5580ff5-at-[206.69.208.22]} } 11, 12 -- Date: Thu, 25 May 2006 17:51:30 -0500 } 11, 12 -- To: microscopy-at-microscopy.com } 11, 12 -- From: mccaulak-at-wfu.edu (by way of } MicroscopyListserver) } 11, 12 -- Subject: viaWWW: procedure for } quantitative fluorescence } 11, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Fri May 26 02:51:58 2006 9, 20 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.87.65]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4Q7pvvo015687 9, 20 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 02:51:57 -0500 9, 20 -- Received: (qmail 85771 invoked by uid 60001); 26 May 2006 07:51:57 -0000 9, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 20 -- s=s1024; d=yahoo.com; 9, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 20 -- b=ClKsXVm9To05z+k4bvYjsr3KQPV/+RHYDE3LV/tEwu92wgmM0nfPTbHDBuswVWVb2Gwe89HAfn3OGNQBpSJxQs+uE9LJ5JSSdZjuYZoWBi5/oLuqCDk4/3bDeFjXuuAEPnK4FJtjHJOF1FVNDTZ6YMPvwkF/PIAyCswEQxGyVOo= ; 9, 20 -- Message-ID: {20060526075157.85769.qmail-at-web37412.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Fri, 26 May 2006 00:51:57 PDT 9, 20 -- Date: Fri, 26 May 2006 00:51:57 -0700 (PDT) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] viaWWW: procedure for quantitative fluorescence 9, 20 -- To: mccaulak-at-wfu.edu 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200605252258.k4PMw833014491-at-ns.microscopy.com} 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; charset=iso-8859-1 9, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I have a triple labeling in fluorescence (green-red-blue), but I take pictures in epifluorescence with a B/W camera, which is more sensitive than the colour camera. Now for presentation purposes I would like to give colours to my B/W pictures in the 3 colours originally used and (perhaps) merge them in the end. What is the procedure in Photoshop?
Stéphane
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==============================Original Headers============================== 4, 18 -- From nizets2-at-yahoo.com Fri May 26 03:28:23 2006 4, 18 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4Q8SM6o026230 4, 18 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 03:28:23 -0500 4, 18 -- Received: (qmail 48820 invoked by uid 60001); 26 May 2006 08:28:22 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.com; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 4, 18 -- b=QBi3fX2S9fpqFjDyYCqdY1V71Z2w7YZDzUyJ4ZQwUakYLfeyv9YOcaHCrIaEWTXHoaJ4Pz3/GPRhHovtG/QhWzHwJ5rbpF2J+MrliqD7Z+pAhjfSHiSUdODmT6Hjk56Q217Z2lUElGM2J85N5RWEpLpiR2ARFahUTXALtfcky/Y= ; 4, 18 -- Message-ID: {20060526082822.48817.qmail-at-web37413.mail.mud.yahoo.com} 4, 18 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Fri, 26 May 2006 01:28:22 PDT 4, 18 -- Date: Fri, 26 May 2006 01:28:22 -0700 (PDT) 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 18 -- Subject: coloring B/W pictures with photoshop 4, 18 -- To: microscopy-at-microscopy.com 4, 18 -- MIME-Version: 1.0 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 4, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
It's a bit of a fag in Photoshop - much easier in MetaMorph or ImagePro Plus (and probably in ImageJ/NIHImage for poor people). Photoshop also gets cross with 12-bit B&W images (they go black and it's not worth bringing the image back with contrast adjustment) so export at 8-bit (256 grey) TIF.
I'll email the complete pdf on 'how to do it' that includes VDU screen dumps. For the listserver the basic text is appended below. I have the Bio-Rad PIC plug-ins for Photoshop if anyone would like them.
Keith ---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
Text from the pdf file 'Colouring B&W pictures in photoshop'
Creating Combined RGB Bio-Rad images in Photoshop
(You may need the Bio-Rad Plugin for *.PIC files otherwise use *.TIF) Load images (via import, Bio-Rad PIC Import) Convert both to RGB (Image, Mode,RGB Color) Find the image size (Image, Image Size – 1024x1024 in this case)
Create a new Image for the blue channel (1024x1024 – black) File, New (White, 1024x1024 – it must be the same size as the red and green images) Select Color Picker (left menu – black/white boxes) and change the foreground colour to black (click OK). Right Click over the new image and select all.
Select Edit,Fill and click OK. Image will now be black (the unused blue channel).
Select Windows, Show Channels (to see RGB channels for all images)
Go to the black (blue) image (right click to select all if necessary, and Edit, Copy Select the Green Channel Image and highlight its blue channel
Select Edit, Paste and the blue channel is ‘removed’ on the target image.
Repeat this pasting the black into the red channel (leaving the Green channel left)
Now right click select all over the ‘red channel’ B&W image. Select Edit, Copy
Reselect the ‘green channel’ image (that has black over its red and blue channels).
Select the blackened red channel in the ‘green’ image. Now Edit, Paste (and the red channel is pasted into the red channel of the green image creating a Red and Green combined RGB image (black for blue).
I also adjusted the Image, Adjust, brightness/Contrast for both the red and green images prior to combining them. You must select the red or green channel when adjusting. Once finished select and copy the red image into the red channel of the green image to combine as before. This time the green brightness/contrast has been reduce and the red increased. See resulting image below and compare with the unadjusted original intensities above.
Author: Dr Keith J Morris, Imaging Facility Manager, Cell Biology Division, the institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL.. Telephone: 020 7608 4050. Email: keith.morris-at-ucl.ac.uk.
Please advise me of any errors found and/or difficulties encountered when using this help document.
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: 26 May 2006 09:33 To: keith.morris-at-ucl.ac.uk
Dear listers,
I have a triple labeling in fluorescence (green-red-blue), but I take pictures in epifluorescence with a B/W camera, which is more sensitive than the colour camera. Now for presentation purposes I would like to give colours to my B/W pictures in the 3 colours originally used and (perhaps) merge them in the end. What is the procedure in Photoshop?
Stéphane
==============================Original Headers============================== 30, 27 -- From keith.morris-at-ucl.ac.uk Fri May 26 03:59:23 2006 30, 27 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 30, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4Q8xNJN004178 30, 27 -- for {Microscopy-at-microscopy.com} ; Fri, 26 May 2006 03:59:23 -0500 30, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 30, 27 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 30, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 30, 27 -- id 1FjYAA-0002GJ-7X 30, 27 -- for Microscopy-at-microscopy.com; Fri, 26 May 2006 09:59:18 +0100 30, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 30, 27 -- To: {Microscopy-at-microscopy.com} 30, 27 -- Subject: RE: [Microscopy] colouring B/W pictures with Photoshop 30, 27 -- Date: Fri, 26 May 2006 09:59:17 +0100 30, 27 -- Message-ID: {000301c680a2$ab5a58c0$7b865290-at-keithhigrade} 30, 27 -- MIME-Version: 1.0 30, 27 -- Content-Type: text/plain; 30, 27 -- charset="iso-8859-1" 30, 27 -- X-Mailer: Microsoft Office Outlook 11 30, 27 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 30, 27 -- Thread-Index: AcaAnvRj1DvV6/0CTNSms6Zt0Pb/DAAARgAw 30, 27 -- In-Reply-To: {200605260832.k4Q8WZNd000976-at-ns.microscopy.com} 30, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 30, 27 -- X-UCL-MailScanner: Found to be clean 30, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 30, 27 -- X-Spam-Status: No 30, 27 -- Content-Transfer-Encoding: 8bit 30, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4Q8xNJN004178 ==============================End of - Headers==============================
It's not really possible to get quantitative measurements of epi-fluorescence. The gel fluorescence standards commonly used are simply to check that your camera/detector and electronics are genuinely linear in response (something quite important to know) - they are no use as standards for calibrating fluorescence within the cell (although you can use things like BCECF in solution at known pH's to roughly calibrate intracellular pH). To properly calibrate cellular fluorescence you need standards that are essentially known concentrations and masses of labelled protein etc.. within cells, not something that's easy to get. You will always have problems with laser power variation, differential bleaching, uneven inconsistent labelling, internal quenching etc.. so you will probably never be able to say that one cell has exactly twice the mass (total pixel brightness) or concentration (mean pixel brightness) of a given protein compared to another. However it is reasonable to assume that a very brightly labelled cell has definitely more labelled fluorochrome in it than a poorly fluorescent cell or intracellular region (and you could say something like 'suggesting' x times concentration etc.). Also try measuring something simple like concentrations of fluorescent beads to see how (badly) the image analysis results work out (these do have the advantage that you can actually count them as well).
You will probably have to use regions of interest to include darker as well as bright areas if comparing regions, plus you can use peak values as described by Stephane (but often the bright sample fluorescence area has to be very clearly defined to one region to get this to work). You can also try things like what % of the cell is brighter than a set grey level. Co-localisation (i.e. the % of bright pixels in both the red and green channel across the cell) is also useful. Generally each set of samples require their own image analysis protocols depending on what you want to find out, plus you need simple stats to say if the difference is significant to 95% level.
When looking at optical density, this can be more easily calibrated - here you are measuring optical white light transmission though a sample. So you can create black (metal disk), white (free space) and have a selection of known density materials like polythene (grey) etc. in the image. You can therefore assign a grey level to a particular density. We have used this to estimate things like bone density, assuming your can get very evenly sliced samples.
Keith ---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
Hello, I've been processing some small plant roots for scanning electron microscopy. I use Glutaraldehyde fixation followed by 0.8% KFerrocyanide and 1% OsO4 - otherwise standard fixation. I get great root preservation, but I've found that the root hairs all look very collapsed. I can send a picture if you like, but was wondering if anyone had experience in this area.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 2, 24 -- From gvrdolja-at-nature.berkeley.edu Thu May 25 18:14:50 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 k4PNEo4t017400 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 25 May 2006 18:14:50 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id AEC69C1E39 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 25 May 2006 16:14:49 -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 25387-03 for {microscopy-at-microscopy.com} ; 2, 24 -- Thu, 25 May 2006 16:14:37 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id 5E8A1C1E57; Thu, 25 May 2006 16:14:37 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 321C1C1E55 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 25 May 2006 16:14:36 -0700 (PDT) 2, 24 -- Date: Thu, 25 May 2006 16:14:36 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: microscopy-at-microscopy.com 2, 24 -- Subject: roots for EM 2, 24 -- Message-ID: {Pine.SOC.4.64.0605251612590.20397-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==============================
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==============================Original Headers============================== 17, 32 -- From David.Patton-at-uwe.ac.uk Fri May 26 05:51:34 2006 17, 32 -- Received: from mailapp02.uwe.ac.uk (mailapp02.uwe.ac.uk [164.11.132.63]) 17, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4QApXUk029014 17, 32 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 05:51:33 -0500 17, 32 -- Received: from (164.11.132.60) by mailapp02.uwe.ac.uk via smtp 17, 32 -- id 1dcf_17b4899a_eca6_11da_99e5_0002b3c90020; 17, 32 -- Fri, 26 May 2006 11:55:10 +0100 17, 32 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 17, 32 -- (egen-uwe01.campus.ads.uwe.ac.uk [164.11.249.121]) 17, 32 -- by mta01.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 17, 32 -- 2005)) with ESMTP id {0IZV00203CTSW9-at-mta01.uwe.ac.uk} for 17, 32 -- microscopy-at-microscopy.com; Fri, 26 May 2006 11:51:28 +0100 (BST) 17, 32 -- Date: Fri, 26 May 2006 11:51:27 +0100 17, 32 -- From: David Patton {David.Patton-at-uwe.ac.uk} 17, 32 -- Subject: RE: [Microscopy] roots for EM 17, 32 -- To: gvrdolja-at-nature.berkeley.edu, microscopy-at-microscopy.com 17, 32 -- Message-id: {F247F674896BE243AD8263C5280E2BDB024A6565-at-egen-uwe01} 17, 32 -- MIME-version: 1.0 17, 32 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.6944.0 17, 32 -- Content-type: text/plain; charset=us-ascii 17, 32 -- Content-class: urn:content-classes:message 17, 32 -- Thread-topic: [Microscopy] roots for EM 17, 32 -- Thread-index: AcaAUSdLjXz6EbVvRuev1AbUO4tscQAYKmTw 17, 32 -- X-MS-Has-Attach: 17, 32 -- X-MS-TNEF-Correlator: 17, 32 -- X-NAI-Spam-Score: -1.2 17, 32 -- X-NAI-Spam-Rules: 1 Rules triggered 17, 32 -- BAYES_01=-1.2 17, 32 -- X-NAIMIME-Disclaimer: 1 17, 32 -- X-NAIMIME-Modified: 1 17, 32 -- Content-Transfer-Encoding: 8bit 17, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4QApXUk029014 ==============================End of - Headers==============================
Have you considered cryo-SEM? This would seem to the ideal method - maintaining the sample in its hydrated state and avoiding mechanical damage that critical point drying can cause some delicate samples.
This is quite an old image and at one time (now lost) there was a companion image showing the clean surface after controlled etching (sublimation) of the ice.
Declaration of commercial interest: Quorum Technologies are manufacturers of the PP2000 and PP2000T cryo-SEM systems.
Best regards,
Mike Wombwell Quorum Technologies Newhaven, East Sussex, UK Tel: +44(0)1273 510535 Fax: +44(0)1273 510536 mike.wombwell-at-quorumtech.com http://www.quorumtech.com E & O E
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Hello, I've been processing some small plant roots for scanning electron microscopy. I use Glutaraldehyde fixation followed by 0.8% KFerrocyanide and 1% OsO4 - otherwise standard fixation. I get great root preservation, but I've found that the root hairs all look very collapsed. I can send a picture if you like, but was wondering if anyone had experience in this area.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \/\/\ 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
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Organization: Charles UNiversity in Prague, Czech Republic
Title-Subject: [Filtered] FDA
Question: Dear listers, some time ago I decided to distinguish between dead and living cells in plant roots using FDA (fluorescein diacetate) combined with PI (propidium iodide). But it seems cells showing the brigtest FDA fluorescence are those that are prone to die, forming strange fluorescing vesicles or even their whole cell content fluoresces. But haven't found enough supporting information in literature. Does any of you have similar experience (or any citations) showing the activity of esterases would more refer to physiological changes (let's say connected with cell death) of the cell than just to its viability? Thanks in advance for your help,
I don't know the real answer to your question but I suspect it's something to do with the toxicity of propidium iodide. Do you use PI and FDA at the same time? PI on it's own is a fairly reliable live/dead marker in Arabidopsis roots when used at 5 micrograms/ml. Living cells exclude it and so remain outlined while dead cells fill with the stain and fluoresce brightly. The problem with PI is that it will kill the cells if they are left too long in it (greater than 30-45 minutes). Maybe that is why the wierd FDA effect. What happens when you use the stains on their own?
Regards, John.
lenoska1-at-seznam.cz wrote:
} Email: lenoska1-at-seznam.cz } Name: Zuzana Lenochova } } Organization: Charles UNiversity in Prague, Czech Republic } } Title-Subject: [Filtered] FDA } } Question: Dear listers, } some time ago I decided to distinguish between dead and living cells in plant roots using FDA (fluorescein diacetate) combined with PI (propidium iodide). But it seems cells showing the brigtest FDA fluorescence are those that are prone to die, forming strange fluorescing vesicles or even their whole cell content fluoresces. But haven't found enough supporting information in literature. Does any of you have similar experience (or any citations) showing the activity of esterases would more refer to physiological changes (let's say connected with cell death) of the cell than just to its viability? } Thanks in advance for your help, } } Zuzana } } }
--
********************************* C. John Runions, Ph.D. School of Biological and Molecular Sciences Oxford Brookes University Oxford, UK OX3 0BP
As Keith implies, it is quite easy to do the kind of work you want with ImageJ. There is a function under the Image Menu} Colors, called RGB merge. Using that, you can assign any b/w image to one of the three color channels, or to none. This does, however, presuppose that your three images are properly aligned. There is also a plugin for ImageJ that will allow you to align the images.
To replace a single b/w image with a colored one, you can use the ImageJ LookUpTables, which will remap the 8-bit gray scale to a color of your choosing.
Check with me if you need more information.
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 } ---------------------------------------------------------------------- } ------ } } Hi Stephanie, } } It's a bit of a fag in Photoshop - much easier in MetaMorph or } ImagePro Plus (and probably in ImageJ/NIHImage for poor people). } Photoshop also gets cross with 12-bit B&W images (they go black and } it's not worth bringing the image back with contrast adjustment) so } export at 8-bit (256 grey) TIF. } } I'll email the complete pdf on 'how to do it' that includes VDU screen } dumps. For the listserver the basic text is appended below. I have the } Bio-Rad PIC plug-ins for Photoshop if anyone would like them. } } Keith } ---------------------------------------------------------- } Dr Keith J Morris } Imaging Facilities Manager } Cell Biology Division } Institute of Ophthalmology } University College London } 11-43 Bath Street } London EC1V 9EL } } Tel: 020 7608 4050 } Fax: 020 7608 4034 } email: keith.morris-at-ucl.ac.uk } } } Text from the pdf file 'Colouring B&W pictures in photoshop' } } Creating Combined RGB Bio-Rad images in Photoshop } } (You may need the Bio-Rad Plugin for *.PIC files otherwise use *.TIF) } Load images (via import, Bio-Rad PIC Import) Convert both to RGB } (Image, Mode,RGB Color) Find the image size (Image, Image Size – } 1024x1024 in this case) } } Create a new Image for the blue channel (1024x1024 – black) } File, New (White, 1024x1024 – it must be the same size as the red and } green images) Select Color Picker (left menu – black/white boxes) and } change the foreground colour to black (click OK). Right Click over the } new image and select all. } } Select Edit,Fill and click OK. Image will now be black (the unused } blue channel). } } Select Windows, Show Channels (to see RGB channels for all images) } } Go to the black (blue) image (right click to select all if necessary, } and Edit, Copy Select the Green Channel Image and highlight its blue } channel } } Select Edit, Paste and the blue channel is ‘removed’ on the target } image. } } Repeat this pasting the black into the red channel (leaving the Green } channel left) } } Now right click select all over the ‘red channel’ B&W image. } Select Edit, Copy } } Reselect the ‘green channel’ image (that has black over its red and } blue channels). } } Select the blackened red channel in the ‘green’ image. } Now Edit, Paste (and the red channel is pasted into the red channel of } the green image creating a Red and Green combined RGB image (black } for blue). } } I also adjusted the Image, Adjust, brightness/Contrast for both the } red and green images prior to combining them. You must select the red } or green channel when adjusting. Once finished select and copy the red } image into the red channel of the green image to combine as before. } This time the green brightness/contrast has been reduce and the red } increased. See resulting image below and compare with the unadjusted } original intensities above. } } Author: Dr Keith J Morris, Imaging Facility Manager, Cell Biology } Division, the institute of Ophthalmology, 11-43 Bath Street, London } EC1V 9EL.. Telephone: 020 7608 4050. Email: } keith.morris-at-ucl.ac.uk. } } Please advise me of any errors found and/or difficulties encountered } when using this help document. } } } } } } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: 26 May 2006 09:33 } To: keith.morris-at-ucl.ac.uk } Subject: [Microscopy] coloring B/W pictures with photoshop } } } } } ---------------------------------------------------------------------- } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Dear listers, } } I have a triple labeling in fluorescence } (green-red-blue), but I take pictures in } epifluorescence with a B/W camera, which is more } sensitive than the colour camera. Now for presentation } purposes I would like to give colours to my B/W } pictures in the 3 colours originally used and } (perhaps) merge them in the end. } What is the procedure in Photoshop? } } Stéphane } } } } ==============================Original } Headers============================== 30, 27 -- From } keith.morris-at-ucl.ac.uk Fri May 26 03:59:23 2006 30, 27 -- Received: } from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 30, 27 } -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k4Q8xNJN004178 30, 27 -- for {Microscopy-at-microscopy.com} ; Fri, 26 May } 2006 03:59:23 -0500 30, 27 -- Received: from cell23.ioo.ucl.ac.uk } ([144.82.134.123] helo=keithhigrade) 30, 27 -- by vscani-e.ucl.ac.uk } with esmtp (Exim 4.60) 30, 27 -- (envelope-from } {keith.morris-at-ucl.ac.uk} ) 30, 27 -- id 1FjYAA-0002GJ-7X 30, 27 -- } for Microscopy-at-microscopy.com; Fri, 26 May 2006 09:59:18 +0100 30, 27 } -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 30, 27 -- To: } {Microscopy-at-microscopy.com} 30, 27 -- Subject: RE: [Microscopy] } colouring B/W pictures with Photoshop 30, 27 -- Date: Fri, 26 May 2006 } 09:59:17 +0100 30, 27 -- Message-ID: } {000301c680a2$ab5a58c0$7b865290-at-keithhigrade} 30, 27 -- MIME-Version: } 1.0 30, 27 -- Content-Type: text/plain; 30, 27 -- } charset="iso-8859-1" 30, 27 -- X-Mailer: Microsoft Office Outlook 11 } 30, 27 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 30, } 27 -- Thread-Index: AcaAnvRj1DvV6/0CTNSms6Zt0Pb/DAAARgAw 30, 27 -- } In-Reply-To: {200605260832.k4Q8WZNd000976-at-ns.microscopy.com} 30, 27 -- } X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, } helpdesk-at-ucl.ac.uk for more information 30, 27 -- X-UCL-MailScanner: } Found to be clean 30, 27 -- X-UCL-MailScanner-From: } keith.morris-at-ucl.ac.uk 30, 27 -- X-Spam-Status: No 30, 27 -- } Content-Transfer-Encoding: 8bit 30, 27 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id k4Q8xNJN004178 } ==============================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============================== 11, 29 -- From jbs-at-temple.edu Fri May 26 08:59:26 2006 11, 29 -- Received: from po-smtp4.temple.edu (po-smtp4.temple.edu [155.247.166.232]) 11, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QDxQ3A007903 11, 29 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 08:59:26 -0500 11, 29 -- Received: from [155.247.98.40] (jbs.bio.temple.edu [155.247.98.40]) 11, 29 -- by po-smtp4.temple.edu (MOS 3.7.5a-GA) 11, 29 -- with ESMTP id CXC77817 (AUTH jbs); 11, 29 -- Fri, 26 May 2006 09:59:25 -0400 (EDT) 11, 29 -- From: "Joel Sheffield" {jbs-at-temple.edu} 11, 29 -- To: microscopy-at-microscopy.com 11, 29 -- Date: Fri, 26 May 2006 09:57:28 -0400 11, 29 -- MIME-Version: 1.0 11, 29 -- Subject: Re: [Microscopy] RE: colouring B/W pictures with Photoshop 11, 29 -- Reply-to: jbs-at-temple.edu 11, 29 -- Message-ID: {4476D108.11056.8C0C14D-at-jbs.temple.edu} 11, 29 -- Priority: normal 11, 29 -- In-reply-to: {200605260859.k4Q8xYIx004391-at-ns.microscopy.com} 11, 29 -- X-mailer: Pegasus Mail for Windows (4.30 public beta 1) 11, 29 -- Content-type: text/plain; charset=ISO-8859-1 11, 29 -- Content-description: Mail message body 11, 29 -- X-Junkmail: UCE(51) 11, 29 -- X-Junkmail-Status: score=51/50, host=po-smtp4.temple.edu 11, 29 -- X-Junkmail-SD-Raw: score=bulk(1), 11, 29 -- refid=str=0001.0A090209.4476C0F5.0032,ss=3,sh,fgs=0, 11, 29 -- ip=155.247.98.40, 11, 29 -- so=2006-05-09 23:27:51, 11, 29 -- dmn=5.2.4/2006-05-04 11, 29 -- Content-Transfer-Encoding: 8bit 11, 29 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k4QDxQ3A007903 ==============================End of - Headers==============================
I would like to transfer 16 bit sun raster files into TIF format without loosing the resolution !. Anybody aware of a technique/software which can do that?
Regards,
Jafar
Rutgers University, Piscataway, NJ 08854 Jafar F. Al-Sharab, Ph.D. Rutgers, The State University of New Jersey Department of Materials Science and Engineering 607 Taylor Road, Room 237 Piscataway, NJ 08854-8065 Tel. 732-445-5615 Work 732-688-1955 Cell Fax 732-445-3258
E-mail: jafarhan-at-rci.rutgers.edu
==============================Original Headers============================== 7, 23 -- From jafarhan-at-rci.rutgers.edu Fri May 26 09:55:31 2006 7, 23 -- Received: from annwn13.rutgers.edu (smtp.rutgers.edu [128.6.72.243]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QEtVl8018990 7, 23 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 09:55:31 -0500 7, 23 -- Received: from localhost (localhost.rutgers.edu [127.0.0.1]) 7, 23 -- by annwn13.rutgers.edu (Postfix) with ESMTP id 77FCC32408B 7, 23 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 10:55:31 -0400 (EDT) 7, 23 -- Received: from annwn13.rutgers.edu ([127.0.0.1]) 7, 23 -- by localhost (annwn13.rutgers.edu [127.0.0.1]) (amavisd-new, port 10024) 7, 23 -- with ESMTP id 06775-07 for {microscopy-at-microscopy.com} ; 7, 23 -- Fri, 26 May 2006 10:55:31 -0400 (EDT) 7, 23 -- Received: from galt.rci.rutgers.edu (jafarhan2.engr.rutgers.edu [128.6.22.35]) 7, 23 -- by annwn13.rutgers.edu (Postfix) with ESMTP id 42F4332408A 7, 23 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 10:55:31 -0400 (EDT) 7, 23 -- Message-Id: {7.0.1.0.0.20060526105426.0193a2d8-at-rci.rutgers.edu} 7, 23 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 7, 23 -- Date: Fri, 26 May 2006 10:55:31 -0400 7, 23 -- To: microscopy-at-microscopy.com 7, 23 -- From: Jafar Al-Sharab {jafarhan-at-rci.rutgers.edu} 7, 23 -- Subject: Transferring sun raster into tif format 7, 23 -- Mime-Version: 1.0 7, 23 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 7, 23 -- X-Virus-Scanned: Virus Scanned by NBCS ==============================End of - Headers==============================
Folks- Can anyone advise us who can service older ultramicrotomes in the Midwest? We have Sorval MT-2 and MT-2B and LKB Ultrotome III ultramicrotomes. Carol Heckman -- __ 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 ___________________________________________________________________________
==============================Original Headers============================== 1, 15 -- From heckman-at-bgnet.bgsu.edu Fri May 26 11:09:24 2006 1, 15 -- Received: from smtp01.bgsu.edu (smtp01.bgsu.edu [129.1.5.17]) 1, 15 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QG9N8d030580 1, 15 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 11:09:24 -0500 1, 15 -- Received: from [129.1.85.81] (dhcp-85-81.bgsu.edu [129.1.85.81]) 1, 15 -- by smtp01.bgsu.edu (Switch-3.1.8/Switch-3.1.6) with ESMTP id k4QG9M0S018033 1, 15 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 12:09:23 -0400 (EDT) 1, 15 -- Mime-Version: 1.0 1, 15 -- X-Sender: heckman-at-mailstore.bgsu.edu (Unverified) 1, 15 -- Message-Id: {p04320403c09cf6e506d6-at-[129.1.85.81]} 1, 15 -- Date: Fri, 26 May 2006 12:09:21 -0700 1, 15 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 1, 15 -- From: Carol Heckman {heckman-at-bgnet.bgsu.edu} 1, 15 -- Subject: service on older model ultramicrotomes 1, 15 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
This is a great email to bring back up the ethics of imaging!
What *color* was really collected? For a merged image the interpretation is great using color as described in the follow up emai. BUT, for single image display on a computer (PDF or Powerpoint) the information content is much more accessible to the viewer as grayscale than when displayed in color.
Why? RGB. When you have a grayscale image on the monitor it uses all three equally to make the image. When you have only the R channel on, the intensity is cut by 1/3. The issue there becomes detectable differences. Its a simple exercise. And I encourage everyone to try it: Take a grayscale image (any will do, assuming it has a close to normal distributed histogram) and then make it one color (just Red for example). For many of you this is a normal and obvious statement.
If you are overlaying color it makes sense to convert to color. If you are collecting an image with a color camera, and it is looking at the color (is it a real color or just the filtered spectrum defined by the bandpass filters), okay. But why reduce the information displayed in a single channel.
Maybe my personal bias against such practices as displaying the single chanel GFP label as green is unjust, but I ask: If displaying as a color makes it more difficult to see the information in the micrograph, Why do it? (suggesting that it is less confusing for the viewer doesn't cut it - that's what figure legends and labels do very successfully).
-Geoff
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Fri 5/26/2006 4:32 AM To: Williams, Geoffrey
Dear listers,
I have a triple labeling in fluorescence (green-red-blue), but I take pictures in epifluorescence with a B/W camera, which is more sensitive than the colour camera. Now for presentation purposes I would like to give colours to my B/W pictures in the 3 colours originally used and (perhaps) merge them in the end. What is the procedure in Photoshop?
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 4, 18 -- From nizets2-at-yahoo.com Fri May 26 03:28:23 2006 4, 18 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4Q8SM6o026230 4, 18 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 03:28:23 -0500 4, 18 -- Received: (qmail 48820 invoked by uid 60001); 26 May 2006 08:28:22 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.com; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 4, 18 -- b=QBi3fX2S9fpqFjDyYCqdY1V71Z2w7YZDzUyJ4ZQwUakYLfeyv9YOcaHCrIaEWTXHoaJ4Pz3/GPRhHovtG/QhWzHwJ5rbpF2J+MrliqD7Z+pAhjfSHiSUdODmT6Hjk56Q217Z2lUElGM2J85N5RWEpLpiR2ARFahUTXALtfcky/Y= ; 4, 18 -- Message-ID: {20060526082822.48817.qmail-at-web37413.mail.mud.yahoo.com} 4, 18 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Fri, 26 May 2006 01:28:22 PDT 4, 18 -- Date: Fri, 26 May 2006 01:28:22 -0700 (PDT) 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 18 -- Subject: coloring B/W pictures with photoshop 4, 18 -- To: microscopy-at-microscopy.com 4, 18 -- MIME-Version: 1.0 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 4, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 18, 30 -- From Geoffrey_Williams-at-brown.edu Fri May 26 11:21:21 2006 18, 30 -- Received: from pyxis.services.brown.edu (pyxis.services.brown.edu [128.148.106.154]) 18, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QGLLcJ008286 18, 30 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 11:21:21 -0500 18, 30 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 18, 30 -- by pyxis.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k4QGLI95019717 18, 30 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 12:21:18 -0400 (EDT) 18, 30 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 18, 30 -- Fri, 26 May 2006 12:21:18 -0400 18, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 18, 30 -- Content-class: urn:content-classes:message 18, 30 -- MIME-Version: 1.0 18, 30 -- Content-Type: text/plain; 18, 30 -- charset="iso-8859-1" 18, 30 -- Subject: RE: [Microscopy] coloring B/W pictures with photoshop 18, 30 -- Date: Fri, 26 May 2006 12:21:18 -0400 18, 30 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F032FD36A-at-MAIL1.AD.Brown.Edu} 18, 30 -- X-MS-Has-Attach: 18, 30 -- X-MS-TNEF-Correlator: 18, 30 -- Thread-Topic: [Microscopy] coloring B/W pictures with photoshop 18, 30 -- Thread-Index: AcaAnuwFVGcNirdkTqCZhP6k/m6woQAP8BCZ 18, 30 -- References: {200605260832.k4Q8WNVh000630-at-ns.microscopy.com} 18, 30 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 18, 30 -- To: {microscopy-at-microscopy.com} 18, 30 -- X-OriginalArrivalTime: 26 May 2006 16:21:18.0824 (UTC) FILETIME=[6B528E80:01C680E0] 18, 30 -- X-Brown-Proofpoint: Not Infected 18, 30 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06050800 definitions=3.0.0-06052603 18, 30 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-06050800 definitions=3.0.0-06052603 18, 30 -- Content-Transfer-Encoding: 8bit 18, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4QGLLcJ008286 ==============================End of - Headers==============================
Dear Stephane, the simplest manner in Photoshop is to open all 3 images then convert 1 to RGB and paste in the other channels. Some acquisition software automatically exports monochrome images as 24-bit RGB, saving you this step. Different steps may be needed for the various formats in which captured images are saved.
In Photoshop, view the Channels window, If you have the "red" image converted to RGB, select its "green" channel, then go to the "green" image, 'Select All', 'Copy', return to the green channel in the target image and 'Paste'. Repeat with the "Blue" channel and you are done. Now, select the "RGB" channel at the top of the Channels window to observe the merged image. Open the Levels command to adjust histogram stretch and gamma for each channel in the merged image (be certain to note gamma changes in methods or caption). If you have 16-bit images, John and Christian Russ have available a plugin to scale from 16-bit to 8-bit.
Regards Glen
Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
On May 26, 2006, at 1:30 AM, nizets2-at-yahoo.com wrote:
} Dear listers, } } I have a triple labeling in fluorescence } (green-red-blue), but I take pictures in } epifluorescence with a B/W camera, which is more } sensitive than the colour camera. Now for presentation } purposes I would like to give colours to my B/W } pictures in the 3 colours originally used and } (perhaps) merge them in the end. } What is the procedure in Photoshop? } } Stéphane
==============================Original Headers============================== 12, 25 -- From glenmac-at-u.washington.edu Fri May 26 12:00:05 2006 12, 25 -- Received: from mxout4.cac.washington.edu (mxout4.cac.washington.edu [140.142.33.19]) 12, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QH045D018987 12, 25 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 12:00:05 -0500 12, 25 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 12, 25 -- by mxout4.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k4QH02IF007548; 12, 25 -- Fri, 26 May 2006 10:00:02 -0700 12, 25 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 12, 25 -- (authenticated authid=glenmac) 12, 25 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k4QH0140031420 12, 25 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT); 12, 25 -- Fri, 26 May 2006 10:00:02 -0700 12, 25 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 25 -- In-Reply-To: {200605260830.k4Q8UQnH028964-at-ns.microscopy.com} 12, 25 -- References: {200605260830.k4Q8UQnH028964-at-ns.microscopy.com} 12, 25 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 12, 25 -- Message-Id: {B3438241-BCE6-49E3-BEF0-80834E39F9D6-at-u.washington.edu} 12, 25 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 12, 25 -- Subject: Re: [Microscopy] coloring B/W pictures with photoshop 12, 25 -- Date: Fri, 26 May 2006 09:59:59 -0700 12, 25 -- To: nizets2-at-yahoo.com, microscopy-at-microscopy.com 12, 25 -- X-Mailer: Apple Mail (2.750) 12, 25 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_POSSIBLE_EXPLOIT_SUBJ 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' 12, 25 -- Content-Transfer-Encoding: 8bit 12, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4QH045D018987 ==============================End of - Headers==============================
Jafar F. Al-Sharab asked: "I would like to transfer 16 bit sun raster files into TIF format without loosing the resolution!"
Jafar, what application generated your 16 bit Sun raster files?
Everything I have seen concerning '.ras' files specifies a header structure that does NOT include a 16 bit per pixel option. For example, the excellent site at the University of Rochester's EE site (http://www.earth.rochester.edu/ees254/gmt/doc/html/GMT_Docs/node119.html) lists the ras_depth member of the header structure as having values "Depth (1, 8, 24, 32 bits) of pixel". Likewise, the SunRasterDecoder at the Code Project site (http://www.codeproject.com/bitmap/SUN_Raster_File_Decoder.asp) has also notes that ras-depth is "Depth (1, 8, 24 or 32 bits) of each pixel".
If there is an updated specification or your application "cheats" in handling the format (for instance using 16 bits of a 24 bit pixel, I suspect most of the free converters will not work.
Jafar, if you send me directly (not to the list, please) a small (e.g. 512x512) 16 bit sun raster file, I'll try the tools I have and let you know if any work...
Best regards, John Minter jrminter_at_rochester.rr.com (replace the _at_ with -at-; I'm hoping to avoid the address harvesting bots and minimize spam. My apologies for the inconvenience)
==============================Original Headers============================== 7, 20 -- From jrminter-at-rochester.rr.com Fri May 26 12:10:28 2006 7, 20 -- Received: from ms-smtp-01.nyroc.rr.com (ms-smtp-01.nyroc.rr.com [24.24.2.55]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QHARFv029035 7, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 26 May 2006 12:10:27 -0500 7, 20 -- Received: from jrminternb (cpe-72-226-251-4.rochester.res.rr.com [72.226.251.4]) 7, 20 -- by ms-smtp-01.nyroc.rr.com (8.13.6/8.13.6) with ESMTP id k4QHAMH1020242 7, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 26 May 2006 13:10:23 -0400 (EDT) 7, 20 -- From: "John Minter" {jrminter-at-rochester.rr.com} 7, 20 -- To: {Microscopy-at-microscopy.com} 7, 20 -- Subject: Re: Converting 16 bit/px Sun raster format images to TIF 7, 20 -- Date: Fri, 26 May 2006 13:10:19 -0400 7, 20 -- Message-ID: {000c01c680e7$44487dc0$6501a8c0-at-jrminternb} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="us-ascii" 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- X-Mailer: Microsoft Office Outlook 11 7, 20 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 7, 20 -- Thread-Index: AcaA50Osheg5Q1jJTRibFDFgWbLAwQ== 7, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
There is a very good FREE Image converter program called Imagemagick which I use on Unix, Linux, & Mac's. It has also been ported to Windoze.
http://www.imagemagick.org/script/index.php
If you go to the above site you will be able to download a version for SUN which should let you do the job.
Nestor Your Friendly Neighborhood SysOp
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
===========================================
==============================Original Headers============================== 9, 16 -- From zaluzec-at-aaem.amc.anl.gov Fri May 26 12:28:29 2006 9, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 9, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4QHSTFT006864 9, 16 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 12:28:29 -0500 9, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 9, 16 -- by aaem.amc.anl.gov (8.12.11.20060308/8.12.10) with ESMTP id k4QHSSVg027961 9, 16 -- for {microscopy-at-microscopy.com} ; Fri, 26 May 2006 12:28:28 -0500 9, 16 -- Mime-Version: 1.0 9, 16 -- Message-Id: {p06110406c09ce803d0fa-at-[146.139.72.105]} 9, 16 -- In-Reply-To: {200605261455.k4QEtWJ4018997-at-ns.microscopy.com} 9, 16 -- References: {200605261455.k4QEtWJ4018997-at-ns.microscopy.com} 9, 16 -- Date: Fri, 26 May 2006 12:28:26 -0500 9, 16 -- To: microscopy-at-microscopy.com 9, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 9, 16 -- Subject: Converting sun raster into tif format 9, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
On Fri, 26 May 2006 11:31:37 -0600, {zaluzec-at-aaem.amc.anl.gov} wrote:
} There is a very good FREE Image converter program called } Imagemagick which I use on Unix, Linux, & Mac's. It } has also been ported to Windoze. } } http://www.imagemagick.org/script/index.php } } If you go to the above site you will be able to download } a version for SUN which should let you do the job.
In addition to the excellent ImageMagick toolset Mr. Zaluzec has just recommended, I'd like to encourage you to investigate the Gimp ( http://www.gimp.org ). It is an open-source project intended to provide functionality very similar to Photoshop. It has been ported to just about everything: I use it daily on Mac OS X (under Apple's X11), and prior to that I used it on both Solaris and Windows 98 and XP.
It is a downright invaluable tool, and would still be invaluable at twice the price. Having said that, it's also free, and free is a property that excuses many faults (even though there aren't many faults to object to here!). Precompiled binaries are available at the Gimp web site as well.
It is well worth checking out, especially for those in academia with limited budgets for image manipulation software. Between ImageMagick and the Gimp, you can do an astonishing amount of work.
Hope that helps!
-skod
-- Scott Griffith ISES-LLC 9745 Steeplechase Drive Franktown, CO 80116 303-660-2541 voice 303-660-2542 fax skod-at-ises-llc.com
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A postdoctoral position is available in Surface Transmission Electron Microscopy at Northwestern University. Work would involve collaborative work on a number of different projects including charge density at surfaces, oxide surface structures, surface plasmonics and the surface structure of fuel cell materials. A strong background in TEM is necessary, including basic knowledge of dynamical diffraction and imaging theory. Experience in plan or profile imaging of surfaces would be a plus, but not a requirement.
To apply, send a short CV with a list of publications (no papers) as well as the names of three referees to L-marks-at-northwestern.edu. Include a brief (one page maximum) description of your strengths as relevant to transmission electron microscopy of surfaces.
----------------------------------------------- Professor Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2220 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 email: L - marks -at- northwestern . edu http://www.numis.northwestern.edu -----------------------------------------------
==============================Original Headers============================== 5, 17 -- From ldmm-at-risc4.numis.northwestern.edu Fri May 26 20:33:56 2006 5, 17 -- Received: from risc4.numis.northwestern.edu (risc4.numis.northwestern.edu [129.105.122.70]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4R1XuZE004235 5, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 26 May 2006 20:33:56 -0500 5, 17 -- Received: from localhost (ldmm-at-localhost) 5, 17 -- by risc4.numis.northwestern.edu (8.9.3 (PHNE_28760_binary)/8.9.3) with ESMTP id UAA21600 5, 17 -- for {microscopy-at-msa.microscopy.com} ; Fri, 26 May 2006 20:33:52 -0500 (CDT) 5, 17 -- Date: Fri, 26 May 2006 20:33:52 -0500 (CDT) 5, 17 -- From: LDM Microscopy {ldmm-at-risc4.numis.northwestern.edu} 5, 17 -- To: microscopy-at-msa.microscopy.com 5, 17 -- Subject: Postdoctoral Opening in Surface Microscopy 5, 17 -- In-Reply-To: {BCD0C258-FF8B-11D8-B1E0-000393D603E6-at-caltech.edu} 5, 17 -- Message-ID: {Pine.GHP.4.63.0605262020210.21597-at-risc4.numis.northwestern.edu} 5, 17 -- References: {6.1.2.0.2.20040903182522.024474f8-at-mail.calweb.com} 5, 17 -- {BCD0C258-FF8B-11D8-B1E0-000393D603E6-at-caltech.edu} 5, 17 -- MIME-Version: 1.0 5, 17 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
Thanks Geoffrey for this remark. In the present case the merging of the 3 colors is meaningful because I look for colocalisation (I will add "or not", to remain ethically objective ;-)), but your remark makes sense. Please could you answer to the following considerations?
- What if you make color photo films of TIFF images to project them (classical slide projectors)? - What if you project a TIFF image with a projector directly coupled to the computer instead of a monitor? - Would it be ethically correct to increase the intensity of one color 3 fold in order to compensate the decrease due to RGB pictures displayed on a monitor?
Stéphane
--- Geoffrey_Williams-at-brown.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This is a great email to bring back up the ethics of } imaging! } } What *color* was really collected? For a merged } image the interpretation is great using color as } described in the follow up emai. BUT, for single } image display on a computer (PDF or Powerpoint) the } information content is much more accessible to the } viewer as grayscale than when displayed in color. } } Why? RGB. When you have a grayscale image on the } monitor it uses all three equally to make the image. } When you have only the R channel on, the intensity } is cut by 1/3. The issue there becomes detectable } differences. Its a simple exercise. And I } encourage everyone to try it: Take a grayscale image } (any will do, assuming it has a close to normal } distributed histogram) and then make it one color } (just Red for example). For many of you this is a } normal and obvious statement. } } If you are overlaying color it makes sense to } convert to color. If you are collecting an image } with a color camera, and it is looking at the color } (is it a real color or just the filtered spectrum } defined by the bandpass filters), okay. But why } reduce the information displayed in a single } channel. } } Maybe my personal bias against such practices as } displaying the single chanel GFP label as green is } unjust, but I ask: } If displaying as a color makes it more difficult to } see the information in the micrograph, Why do it? } (suggesting that it is less confusing for the viewer } doesn't cut it - that's what figure legends and } labels do very successfully). } } -Geoff } } } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Fri 5/26/2006 4:32 AM } To: Williams, Geoffrey } Subject: [Microscopy] coloring B/W pictures with } photoshop } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } I have a triple labeling in fluorescence } (green-red-blue), but I take pictures in } epifluorescence with a B/W camera, which is more } sensitive than the colour camera. Now for } presentation } purposes I would like to give colours to my B/W } pictures in the 3 colours originally used and } (perhaps) merge them in the end. } What is the procedure in Photoshop? } } Stéphane } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam } protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 4, 18 -- From nizets2-at-yahoo.com Fri May 26 03:28:23 } 2006 } 4, 18 -- Received: from web37413.mail.mud.yahoo.com } (web37413.mail.mud.yahoo.com [209.191.87.66]) } 4, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k4Q8SM6o026230 } 4, 18 -- for {microscopy-at-microscopy.com} ; Fri, 26 } May 2006 03:28:23 -0500 } 4, 18 -- Received: (qmail 48820 invoked by uid } 60001); 26 May 2006 08:28:22 -0000 } 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; } c=nofws; } 4, 18 -- s=s1024; d=yahoo.com; } 4, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 4, 18 -- } b=QBi3fX2S9fpqFjDyYCqdY1V71Z2w7YZDzUyJ4ZQwUakYLfeyv9YOcaHCrIaEWTXHoaJ4Pz3/GPRhHovtG/QhWzHwJ5rbpF2J+MrliqD7Z+pAhjfSHiSUdODmT6Hjk56Q217Z2lUElGM2J85N5RWEpLpiR2ARFahUTXALtfcky/Y= } ; } 4, 18 -- Message-ID: } {20060526082822.48817.qmail-at-web37413.mail.mud.yahoo.com} } 4, 18 -- Received: from [80.122.101.102] by } web37413.mail.mud.yahoo.com via HTTP; Fri, 26 May } 2006 01:28:22 PDT } 4, 18 -- Date: Fri, 26 May 2006 01:28:22 -0700 (PDT) } 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 4, 18 -- Subject: coloring B/W pictures with } photoshop } 4, 18 -- To: microscopy-at-microscopy.com } 4, 18 -- MIME-Version: 1.0 } 4, 18 -- Content-Type: text/plain; } charset=iso-8859-1 } 4, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== } } } } } ==============================Original } Headers============================== } 18, 30 -- From Geoffrey_Williams-at-brown.edu Fri May } 26 11:21:21 2006 } 18, 30 -- Received: from pyxis.services.brown.edu } (pyxis.services.brown.edu [128.148.106.154]) } 18, 30 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k4QGLLcJ008286 } 18, 30 -- for {microscopy-at-microscopy.com} ; Fri, 26 } May 2006 11:21:21 -0500 } 18, 30 -- Received: from } ex-gateway2-out.AD.Brown.Edu } (ex-gateway2.ad.brown.edu [128.148.21.53]) } 18, 30 -- by pyxis.services.brown.edu } (Switch-3.1.8/Switch-3.1.7/) with ESMTP id } k4QGLI95019717 } 18, 30 -- for {microscopy-at-microscopy.com} ; Fri, 26 } May 2006 12:21:18 -0400 (EDT) } 18, 30 -- Received: from mail1.AD.Brown.Edu } ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu } with Microsoft SMTPSVC(6.0.3790.1830); } 18, 30 -- Fri, 26 May 2006 12:21:18 -0400 } 18, 30 -- X-MimeOLE: Produced By Microsoft Exchange } V6.5.7226.0 } 18, 30 -- Content-class: urn:content-classes:message } 18, 30 -- MIME-Version: 1.0 } 18, 30 -- Content-Type: text/plain; } 18, 30 -- charset="iso-8859-1" } 18, 30 -- Subject: RE: [Microscopy] coloring B/W } pictures with photoshop } 18, 30 -- Date: Fri, 26 May 2006 12:21:18 -0400 } 18, 30 -- Message-ID: } {A1A84D541C161C4C988B4E0FB38F5A0F032FD36A-at-MAIL1.AD.Brown.Edu} } 18, 30 -- X-MS-Has-Attach: } 18, 30 -- X-MS-TNEF-Correlator: } 18, 30 -- Thread-Topic: [Microscopy] coloring B/W } pictures with photoshop } 18, 30 -- Thread-Index: } AcaAnuwFVGcNirdkTqCZhP6k/m6woQAP8BCZ } 18, 30 -- References: } {200605260832.k4Q8WNVh000630-at-ns.microscopy.com} } 18, 30 -- From: "Williams, Geoffrey" } {Geoffrey_Williams-at-brown.edu} } 18, 30 -- To: {microscopy-at-microscopy.com} } 18, 30 -- X-OriginalArrivalTime: 26 May 2006 } 16:21:18.0824 (UTC) FILETIME=[6B528E80:01C680E0] } 18, 30 -- X-Brown-Proofpoint: Not Infected } 18, 30 -- X-Proofpoint-Spam-Details: rule=notspam } policy=default score=0 mlx=-1 adultscore=0 adjust=0 } reason=safe engine=3.1.0-06050800 } definitions=3.0.0-06052603 } 18, 30 -- X-Brown-MailScanner-SpamCheck: not spam, } rule=notspam policy=default score=0 mlx=-1 } adultscore=0 adjust=0 reason=safe } engine=3.1.0-06050800 definitions=3.0.0-06052603 } 18, 30 -- Content-Transfer-Encoding: 8bit } 18, 30 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } k4QGLLcJ008286 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Mon May 29 09:50:57 2006 9, 20 -- Received: from web37407.mail.mud.yahoo.com (web37407.mail.mud.yahoo.com [209.191.87.60]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4TEoveO012135 9, 20 -- for {microscopy-at-microscopy.com} ; Mon, 29 May 2006 09:50:57 -0500 9, 20 -- Received: (qmail 58699 invoked by uid 60001); 29 May 2006 14:50:57 -0000 9, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 20 -- s=s1024; d=yahoo.com; 9, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 20 -- b=lc1grLVtKxAY/wVkdLXRj7DRP8QKYhA0xfzBUWlAzZMUElnI1JxVMqrPXR5RxTvlg9+SN7n12t+aTf+nmGtI/m3X1tmlvT7M7QlERrxog5f1zHORPp1YMxOYsBpcfb7dloMULSFEZfIlmjSf0XyHtXvKZezRknAPgttQx1fJGqM= ; 9, 20 -- Message-ID: {20060529145057.58697.qmail-at-web37407.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37407.mail.mud.yahoo.com via HTTP; Mon, 29 May 2006 07:50:56 PDT 9, 20 -- Date: Mon, 29 May 2006 07:50:56 -0700 (PDT) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] RE: coloring B/W pictures with photoshop 9, 20 -- To: Geoffrey_Williams-at-brown.edu 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200605261625.k4QGPVQO017119-at-ns.microscopy.com} 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; charset=iso-8859-1 9, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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 DrJohnRuss-at-AOL.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: DrJohnRuss-at-AOL.com Name: John Russ
Organization: North Carolina State University
Title-Subject: [Filtered] RE: colouring B/W pictures with Photoshop
Question:
-----Original Message----- } From: keith.morris-at-ucl.ac.uk To: DrJohnRuss-at-AOL.com Sent: Fri, 26 May 2006 03:59:50 -0500
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Email: Amr_elsirafy-at-hotmail.com Name: afaf Ibrahim Mahdi
Organization: Cairo university
Title-Subject: [Filtered] ESEM-detectors
Question: Hello I' d like to know is GSED is used in ESEM with tungesten filament and if it can be used in both low and high vacuum mode because the BSD in my XL30 TMP instrument is damaged and I want to know iif it is best to buy back scatter detectors or GSED and is it need a new software.
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Email: eric.leroy-at-glvt-cnrs.fr Name: Eric Leroy
Organization: CNRS - LCMTR
Title-Subject: [Filtered] AnalySIS TIFF images
Question: Hi,
We have a SIS Megaview III camera attached to our Tecnai and we use AnalySIS to record the images. Since we only have a full version of AnalySIS installed on the microscope's PC we usually treat ours images with ImageJ. ImageJ can perfectly read the TIFF images but is unable to read the spatial calibration of the image. Do you know how this spatial calibration is written in the image ?
I Googled GSED and couldn't find anything related to SEM, so for those of us who are acronym-challenged, could you (or someone else on the list) please define GSED?
Thanks, Bryan Bandli MVA Scientific Consultants
Amr_elsirafy-at-hotmail.com wrote:
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GSED = Gaseous Secondary Electron Detector. It is the secondary electron detector of the ESEM which works at poor vacuum unlike a conventional SE detector.
Dave
-----Original Message----- X-from: bbandli-at-mvainc.com [mailto:bbandli-at-mvainc.com] Sent: 30 May 2006 14:54 To: David Patton
I Googled GSED and couldn't find anything related to SEM, so for those of us who are acronym-challenged, could you (or someone else on the list) please define GSED?
Thanks, Bryan Bandli MVA Scientific Consultants
Amr_elsirafy-at-hotmail.com wrote:
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==============================Original Headers============================== 23, 33 -- From David.Patton-at-uwe.ac.uk Tue May 30 09:06:49 2006 23, 33 -- Received: from mailapp01.uwe.ac.uk (mailapp01.uwe.ac.uk [164.11.132.61]) 23, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4UE6mQS013108 23, 33 -- for {microscopy-at-microscopy.com} ; Tue, 30 May 2006 09:06:48 -0500 23, 33 -- Received: from (164.11.132.62) by mailapp01.uwe.ac.uk via smtp 23, 33 -- id 216a_1a69e016_efe6_11da_8580_0002b3c946e4; 23, 33 -- Tue, 30 May 2006 15:10:52 +0100 23, 33 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 23, 33 -- (egen-uwe01.campus.ads.uwe.ac.uk [164.11.249.121]) 23, 33 -- by mta02.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 23, 33 -- 2005)) with ESMTP id {0J0300HCZ0JBNC-at-mta02.uwe.ac.uk} for 23, 33 -- microscopy-at-microscopy.com; Tue, 30 May 2006 15:06:47 +0100 (BST) 23, 33 -- Date: Tue, 30 May 2006 15:06:46 +0100 23, 33 -- From: David Patton {David.Patton-at-uwe.ac.uk} 23, 33 -- Subject: RE: [Microscopy] Re: viaWWW: ESEM-detectors 23, 33 -- To: bbandli-at-mvainc.com 23, 33 -- Cc: microscopy-at-microscopy.com 23, 33 -- Message-id: {F247F674896BE243AD8263C5280E2BDB024A65F2-at-egen-uwe01} 23, 33 -- MIME-version: 1.0 23, 33 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.6944.0 23, 33 -- Content-type: text/plain; charset=us-ascii 23, 33 -- Content-class: urn:content-classes:message 23, 33 -- Thread-topic: [Microscopy] Re: viaWWW: ESEM-detectors 23, 33 -- Thread-index: AcaD8HSuTGcaiN+ET6q21QQ/yNZLgwAAXNiw 23, 33 -- X-MS-Has-Attach: 23, 33 -- X-MS-TNEF-Correlator: 23, 33 -- X-NAI-Spam-Score: -2.5 23, 33 -- X-NAI-Spam-Rules: 1 Rules triggered 23, 33 -- BAYES_00=-2.5 23, 33 -- X-NAIMIME-Disclaimer: 1 23, 33 -- X-NAIMIME-Modified: 1 23, 33 -- Content-Transfer-Encoding: 8bit 23, 33 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4UE6mQS013108 ==============================End of - Headers==============================
You cannot use GSED in high vac; it is only for low vac. I have a XL30 ESEM and never use the backscatter detector (BSD) since installation six years ago for morphological study, except using it to compare with GSED at the very beginning. A GSED produces images a lot better than a BSD. If I were you, the pick would have been a GSED. If you need a BSD for another reason, then that is another matter to consider. As for software, you don't have to worry; just put in a new GSED and off you go. I have checked with my service engineer.
Ann Fook Yang EM Unit/ Unite EM Bldg 20, AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 yanga-at-agr.gc.ca
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: Amr_elsirafy-at-hotmail.com [mailto:Amr_elsirafy-at-hotmail.com] Sent: Tuesday, May 30, 2006 9:21 AM To: Yang, Ann-Fook
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Email: Amr_elsirafy-at-hotmail.com Name: afaf Ibrahim Mahdi
Organization: Cairo university
Title-Subject: [Filtered] ESEM-detectors
Question: Hello I' d like to know is GSED is used in ESEM with tungesten filament and if it can be used in both low and high vacuum mode because the BSD in my XL30 TMP instrument is damaged and I want to know iif it is best to buy back scatter detectors or GSED and is it need a new software.
The TIFF standard allows for spatial resolution data to be saved in the fields XResolution (no of pixels per measurement unit in X dir) and YResolution (no of pixels per measurement unit in Y dir). The measurement unit data is contained in the ResolutionUnit field (usually inch or cm). From these you should be able to calculate the pixel size, magn or image width. There are a number of free TIFF tag viewers available, for example, http://www.awaresystems.be/imaging/tiff/astifftagviewer.html, or just Google 'tiff tag viewer' . Hope this helps.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
} Name: Eric Leroy } } Organization: CNRS - LCMTR } } Title-Subject: [Filtered] AnalySIS TIFF images } } Question: Hi, } } We have a SIS Megaview III camera attached to our Tecnai and we use } AnalySIS to record the images. Since we only have a full version of } AnalySIS installed on the microscope's PC we usually treat ours images } with ImageJ. ImageJ can perfectly read the TIFF images but is unable to } read the spatial calibration of the image. Do you know how this spatial } calibration is written in the image ? } } Thanks } } Eric } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 9, 12 -- From zaluzec-at-microscopy.com Tue May 30 08:08:56 2006 } 9, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 9, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k4UD8s0s005952 } 9, 12 -- for {microscopy-at-microscopy.com} ; Tue, 30 May 2006 08:08:56 } -0500 } 9, 12 -- Mime-Version: 1.0 } 9, 12 -- X-Sender: (Unverified) } 9, 12 -- Message-Id: {p06110404c0a1f45139b1-at-[206.69.208.22]} } 9, 12 -- Date: Tue, 30 May 2006 08:08:49 -0500 } 9, 12 -- To: microscopy-at-microscopy.com } 9, 12 -- From: eric.leroy-at-glvt-cnrs.fr (by way of MicroscopyListserver) } 9, 12 -- Subject: viaWWW: AnalySIS TIFF images } 9, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers==============================
==============================Original Headers============================== 7, 25 -- From djv23-at-cam.ac.uk Tue May 30 10:52:10 2006 7, 25 -- Received: from ppsw-9.csi.cam.ac.uk (ppsw-9.csi.cam.ac.uk [131.111.8.139]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4UFqAmC002935 7, 25 -- for {microscopy-at-microscopy.com} ; Tue, 30 May 2006 10:52:10 -0500 7, 25 -- X-Cam-SpamDetails: Not scanned 7, 25 -- X-Cam-AntiVirus: No virus found 7, 25 -- X-Cam-ScannerInfo: http://www.cam.ac.uk/cs/email/scanner/ 7, 25 -- Received: from focus.msm.cam.ac.uk ([131.111.100.73]:35996 helo=msm.cam.ac.uk) 7, 25 -- by ppsw-9.csi.cam.ac.uk (ppsw.cam.ac.uk [131.111.8.139]:25) 7, 25 -- with esmtp id 1Fl6Vn-0005eo-T5 (Exim 4.54) 7, 25 -- (return-path {djv23-at-cam.ac.uk} ); Tue, 30 May 2006 16:52:03 +0100 7, 25 -- Received: from davids-pc.cam.ac.uk (sem-djv.msm.cam.ac.uk [131.111.100.208]) 7, 25 -- by msm.cam.ac.uk (8.13.6/8.13.6) with ESMTP id k4UFq2kI010086; 7, 25 -- Tue, 30 May 2006 16:52:02 +0100 (BST) 7, 25 -- Message-Id: {6.2.1.2.0.20060530164338.03c87c20-at-focus.msm.cam.ac.uk} 7, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 7, 25 -- Date: Tue, 30 May 2006 16:58:46 +0100 7, 25 -- To: eric.leroy-at-glvt-cnrs.fr 7, 25 -- From: David Vowles {djv23-at-cam.ac.uk} 7, 25 -- Subject: Re: AnalySIS TIFF images 7, 25 -- Cc: Microscopy Listserver {microscopy-at-microscopy.com} 7, 25 -- In-Reply-To: {200605301315.k4UDFVhF020069-at-ns.microscopy.com} 7, 25 -- References: {200605301315.k4UDFVhF020069-at-ns.microscopy.com} 7, 25 -- Mime-Version: 1.0 7, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
We have the same system. The easiest way to keep the magnification in my pictures I found is to "burn" the scale bar into the image (on your screen it is just in overlay) before saving the picture in jpg or TIFF format. It is a little bit a pain, I would appreciate a method to automatize it, but I don't know if it is possible (I work only for 8 months with this system). It is clear that the pixels "behind" the scale bar will be burned and the information lost.
- Choose "Image" in the Menu bar --} Scale bar... --} draw into overlay Then save your image
Sorry if I misunderstood your question and did not answer it (ask it in french it will be clearer ;-)).
Stéphane
--- eric.leroy-at-glvt-cnrs.fr wrote:
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==============================Original Headers============================== 11, 20 -- From nizets2-at-yahoo.com Wed May 31 01:51:14 2006 11, 20 -- Received: from web37403.mail.mud.yahoo.com (web37403.mail.mud.yahoo.com [209.191.87.56]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4V6pEpq032310 11, 20 -- for {microscopy-at-microscopy.com} ; Wed, 31 May 2006 01:51:14 -0500 11, 20 -- Received: (qmail 63416 invoked by uid 60001); 31 May 2006 06:51:13 -0000 11, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 20 -- s=s1024; d=yahoo.com; 11, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 11, 20 -- b=Uqeg5EcA+8wTdckinu1w0OL8cJxuCh94yqdC7cboyPQ9jzQdo7qRHQ7x3xtyRgmHzWhxYEWXCkPmdbwDqe43X1NVzCGB9UKqU82juRa9cJJWdVDmpIucyvQkN3hfpzltaiB3UMLON5S9Z2EpQk2t9x44O7fbAo/MNezMekLQsdM= ; 11, 20 -- Message-ID: {20060531065113.63414.qmail-at-web37403.mail.mud.yahoo.com} 11, 20 -- Received: from [80.122.101.102] by web37403.mail.mud.yahoo.com via HTTP; Tue, 30 May 2006 23:51:13 PDT 11, 20 -- Date: Tue, 30 May 2006 23:51:13 -0700 (PDT) 11, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 11, 20 -- Subject: Re: [Microscopy] viaWWW: AnalySIS TIFF images 11, 20 -- To: eric.leroy-at-glvt-cnrs.fr 11, 20 -- Cc: microscopy-at-microscopy.com 11, 20 -- In-Reply-To: {200605301320.k4UDKRhG030683-at-ns.microscopy.com} 11, 20 -- MIME-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=iso-8859-1 11, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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 } ---------------------------------------------------------------------------- } } Salut Eric, } } We have the same system. The easiest way to keep the } magnification in my pictures I found is to "burn" the } scale bar into the image (on your screen it is just in } overlay) before saving the picture in jpg or TIFF } format. It is a little bit a pain, I would appreciate } a method to automatize it, but I don't know if it is } possible (I work only for 8 months with this system). } If you save to TIFF file format there is an option you can select in the "Save..." dialog to burn the overlay into the image data. (The other option is to convert to 8bit TIFF file, which sometimes may be helpful, too.)
-at-Eric: There is a viewer that can be downloaded from the SIS webpage[1], which can handle the image files (and databases) created with the full version. This viewer also shows you the data listed in the TIFF header.
Is it possible to use a immunogold purpose silver enhancement kit for revealing other heavy metals, such as cadmium, on paraffin or resin sections?
Best wishes
Fabio
Dr. Fabio D'Amico Dept. Biomedical Sciences University of Catania Via Androne 87/a I-95124 Catania ITALY tel/fax +39 095312017 email: f.damico-at-unict.it
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Thank you for the information. I downloaded the software you told me but unfortunately, the information written int Xresolution tag is allways 200 and the ResolutionUnit is 0. So it seems that the saptial calibration is stored elsewhere.
Best regards
Eric
Le 30/05/06 17:58, « David Vowles » {djv23-at-cam.ac.uk} a écrit :
} Eric, } } The TIFF standard allows for spatial resolution data to be saved in the } fields XResolution (no of pixels per measurement unit in X dir) and } YResolution (no of pixels per measurement unit in Y dir). The measurement } unit data is contained in the ResolutionUnit field (usually inch or cm). } From these you should be able to calculate the pixel size, magn or image } width. There are a number of free TIFF tag viewers available, for example, } http://www.awaresystems.be/imaging/tiff/astifftagviewer.html, or just } Google 'tiff tag viewer' . } Hope this helps. } } David Vowles } Electron Microscope Unit } Dept of Materials Science and Metallurgy } University of Cambridge } Pembroke St Cambridge } UK CB2 3QZ } Tel: +44 (0)1223 334325 } Fax: +44 (0)1223 334567 } Email: djv23-at-cam.ac.uk } } } } } Name: Eric Leroy } } } } Organization: CNRS - LCMTR } } } } Title-Subject: [Filtered] AnalySIS TIFF images } } } } Question: Hi, } } } } We have a SIS Megaview III camera attached to our Tecnai and we use } } AnalySIS to record the images. Since we only have a full version of } } AnalySIS installed on the microscope's PC we usually treat ours images } } with ImageJ. ImageJ can perfectly read the TIFF images but is unable to } } read the spatial calibration of the image. Do you know how this spatial } } calibration is written in the image ? } } } } Thanks } } } } Eric } } } } --------------------------------------------------------------------------- } } } } ==============================Original Headers============================== } } 9, 12 -- From zaluzec-at-microscopy.com Tue May 30 08:08:56 2006 } } 9, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } } 9, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } } id k4UD8s0s005952 } } 9, 12 -- for {microscopy-at-microscopy.com} ; Tue, 30 May 2006 08:08:56 } } -0500 } } 9, 12 -- Mime-Version: 1.0 } } 9, 12 -- X-Sender: (Unverified) } } 9, 12 -- Message-Id: {p06110404c0a1f45139b1-at-[206.69.208.22]} } } 9, 12 -- Date: Tue, 30 May 2006 08:08:49 -0500 } } 9, 12 -- To: microscopy-at-microscopy.com } } 9, 12 -- From: eric.leroy-at-glvt-cnrs.fr (by way of MicroscopyListserver) } } 9, 12 -- Subject: viaWWW: AnalySIS TIFF images } } 9, 12 -- Content-Type: text/plain; charset="us-ascii" } } ==============================End of - Headers============================== }
Eric LEROY Dr. Laboratoire de Chimie Metallurgique des Terres Rares UPR 209 - CNRS Groupe des Laboratoires de Thiais 2-8, rue Henri Dunant 94320 THIAIS cedex
The problem of Eric brings a new question to me: The TEM pictures may be saved in JPG or TIFF formats. I know that TIFF format keeps all the information without compression. But does JPG without compression (quality 12) deteriorates the quality of the picture? I thought that JPG without compression was as good as TIFF.
Stephane
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==============================Original Headers============================== 4, 18 -- From nizets2-at-yahoo.com Wed May 31 07:31:48 2006 4, 18 -- Received: from web37414.mail.mud.yahoo.com (web37414.mail.mud.yahoo.com [209.191.87.67]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4VCVlda014756 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 31 May 2006 07:31:47 -0500 4, 18 -- Received: (qmail 79599 invoked by uid 60001); 31 May 2006 12:31:47 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.com; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 4, 18 -- b=OR+0f0jaY3NylTkKHJTgGTfxY3WS4kLrGgRmUZ+3VSncfFVMPDvyj1ZnnxyocY+3mH9BSMuQQ9vtla/UCLW81Yy8dCdwp9bsESZ8PVDCoKZF9jVntYwHDeYB3SsLEYlQfixGwGwS4jBQxGeBRgpLub2T5Zxfcra0ICLmIxsbvmE= ; 4, 18 -- Message-ID: {20060531123147.79597.qmail-at-web37414.mail.mud.yahoo.com} 4, 18 -- Received: from [80.122.101.102] by web37414.mail.mud.yahoo.com via HTTP; Wed, 31 May 2006 05:31:47 PDT 4, 18 -- Date: Wed, 31 May 2006 05:31:47 -0700 (PDT) 4, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 18 -- Subject: JPG and TIFF formats 4, 18 -- To: microscopy-at-microscopy.com 4, 18 -- MIME-Version: 1.0 4, 18 -- Content-Type: text/plain; charset=iso-8859-1 4, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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Email: lin.wenlang-at-mayo.edu Name: WEN-LANG LIN
Organization: Mayo clinic
Title-Subject: [Filtered] perforated paper labels for embedding molds
Question: Where can I buy some? A full page of 5 1/4" x 7 3/4" was perforated into a total of 7x31 labels, each label is 3/4" x 1/8" and fits one BEEM or gelatin capsules. The full pages are bound like a notepad.
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Email: rpowell-at-nanoprobes.com Name: Rick Powell
Question: [Commercial disclaimer - we make silver enhancement reagents]
Hello Fabio:
Silver enhancement will work on other metals - in our experience, particularly those with an active redox chemistry, or those that are present as nanoparticles.
A good name to search is Gorm Danscher, who is one of the originators of the silver enhancementmethod and has used this, and similar chemistry, to reveal other heavy metals in tissue section (search "Danscher G" on PubMed). I couldn't find any references to cadmium, but he has published several papers on the autometallographic tracing of zinc, which is right above cadmium in the periodic table and would be expected to have similar chemistry:
Danscher, G., and Stoltenberg, M.: Zinc-specific Autometallographic In Vivo Selenium Methods: Tracing of Zinc-enriched (ZEN) Terminals, ZEN Pathways, and Pools of Zinc Ions in a Multitude of Other ZEN Cells. J. Histochem. Cytochem., 53141-153 (2005).
Other papers describe the histochemical tracing of mercury and bismuth using similar techniques.
Hope this helps,
Rick Powell
Dear all,
Is it possible to use a immunogold purpose silver enhancement kit for revealing other heavy metals, such as cadmium, on paraffin or resin sections?
Best wishes
Fabio
Dr. Fabio D'Amico Dept. Biomedical Sciences University of Catania Via Androne 87/a I-95124 Catania ITALY tel/fax +39 095312017 email: f.damico-at-unict.it
***************************************************************************************** Richard D. Powell rpowell-at-nanoprobes.com * www.nanoprobes.com
NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA *****************************************************************************************
Please be advised that there is no implementation of JPG with lossless compression anywhere in the world that I know of. The last time I saw a program that actually was lossless was in DOS 3.3 almost twenty years ago.
Saving with JPG at a setting of 12 removes data. period
If you load an image and make adjustments and then resave it, the image file will have compression on top of compression. The situation gets worse and worse. JPG is an evil format that is not supported for scientific imaging by the Society for really good reasons. If a system produces a different format, the first time you save it should ALWAYS be to tiff if you want to save it.
JPG is for email and web pages and is good for nothing else
Luckily I don't have strong opinions
John
==============================Original Headers============================== 8, 19 -- From john_mackenzie-at-ncsu.edu Wed May 31 08:59:49 2006 8, 19 -- Received: from uni08mr.unity.ncsu.edu (uni08mr.unity.ncsu.edu [152.1.224.167]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4VDxnPJ014387 8, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 31 May 2006 08:59:49 -0500 8, 19 -- Received: from [152.1.178.40] (cord2.emc.ncsu.edu [152.1.178.40]) 8, 19 -- by uni08mr.unity.ncsu.edu (8.13.6/8.13.4/Nv5.2006.0214) with ESMTP id k4VDxlxT006203 8, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 31 May 2006 09:59:48 -0400 (EDT) 8, 19 -- Message-ID: {447DA0EE.2000805-at-ncsu.edu} 8, 19 -- Date: Wed, 31 May 2006 09:58:06 -0400 8, 19 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 8, 19 -- User-Agent: Thunderbird 1.5.0.2 (Windows/20060308) 8, 19 -- MIME-Version: 1.0 8, 19 -- To: Microscopy-at-microscopy.com 8, 19 -- Subject: Re: JPG and TIFF formats 8, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 19 -- Content-Transfer-Encoding: 7bit 8, 19 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.5.31.63507 8, 19 -- X-Spam-Status: No, Hits=7% 8, 19 -- X-Spam-Level: IIIIIII ==============================End of - Headers==============================
At 02:25 AM 5/31/2006, h.dierke-at-tu-braunschweig.de wrote:
} If you save to TIFF file format there is an option you can select in the } "Save..." dialog to burn the overlay into the image data. (The other } option is to convert to 8bit TIFF file, which sometimes may be helpful, } too.)
On our AnalySIS-based systems, "burning in" the overlay also automatically converts the file to 8 bit. The original quantitative image data is lost in the conversion. This gave us fits a few years ago until we found out what was gong on. If I may personify a little, it seems like AnalySIS desperately wants to convert images to 8 bit - the only sure strategy we have found for preserving the 16 bit images is to save them immediately after acquisition, then save again under a different name after any manipulation in AnalySIS.
I should also mention that we are several versions out of date on the software; newer versions of AnalySIS / iTEM / whatever may have changed this behavior.
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
==============================Original Headers============================== 8, 24 -- From voyles-at-engr.wisc.edu Wed May 31 09:20:47 2006 8, 24 -- Received: from mail.cae.wisc.edu (mail.cae.wisc.edu [144.92.13.31]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4VEKl98024778 8, 24 -- for {microscopy-at-microscopy.com} ; Wed, 31 May 2006 09:20:47 -0500 8, 24 -- Received: from portkey.cae.wisc.edu (portkey.cae.wisc.edu [144.92.13.118]) 8, 24 -- by mail.cae.wisc.edu (8.13.3+Sun/8.13.4) with ESMTP id k4VEKZT9014972 8, 24 -- for {microscopy-at-microscopy.com} ; Wed, 31 May 2006 09:20:35 -0500 (CDT) 8, 24 -- Received: from voyles-laptop.engr.wisc.edu (voyles_office2.msae.wisc.edu [128.104.200.162]) 8, 24 -- (authenticated bits=0) 8, 24 -- by portkey.cae.wisc.edu (8.13.4/8.13.4/Debian-3) with ESMTP id k4VEKZaj018408 8, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 8, 24 -- for {microscopy-at-microscopy.com} ; Wed, 31 May 2006 09:20:35 -0500 8, 24 -- Message-Id: {6.2.1.2.2.20060531091110.022a74b8-at-cae.wisc.edu} 8, 24 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 8, 24 -- Date: Wed, 31 May 2006 09:20:29 -0500 8, 24 -- To: microscopy-at-microscopy.com 8, 24 -- From: Paul Voyles {voyles-at-engr.wisc.edu} 8, 24 -- Subject: Re: [Microscopy] viaWWW: AnalySIS TIFF images 8, 24 -- In-Reply-To: {200605310725.k4V7PeKw012665-at-ns.microscopy.com} 8, 24 -- References: {200605310725.k4V7PeKw012665-at-ns.microscopy.com} 8, 24 -- Mime-Version: 1.0 8, 24 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 24 -- X-CAE-MailScanner-Information: Please contact security-at-engr.wisc.edu if this message contains a virus or has been corrupted in delivery. 8, 24 -- X-CAE-MailScanner: Found to be clean (benji) ==============================End of - Headers==============================
Good one John! Now I have an image of Buffy the JPEG Slayer working to save us all from the 'evil JPG.' John.
john_mackenzie-at-ncsu.edu wrote:
} } Stephanie: } } Please be advised that there is no implementation of JPG with lossless } compression anywhere in the world that I know of. The last time I saw a } program that actually was lossless was in DOS 3.3 almost twenty years ago. } } Saving with JPG at a setting of 12 removes data. period } } If you load an image and make adjustments and then resave it, the image } file will have compression on top of compression. The situation gets } worse and worse. } JPG is an evil format that is not supported for scientific imaging by } the Society for really good reasons. If a system produces a different } format, the first time you save it should ALWAYS be to tiff if you want } to save it. } } JPG is for email and web pages and is good for nothing else } } Luckily I don't have strong opinions } } John } } } }
--
********************************* C. John Runions, Ph.D. School of Biological and Molecular Sciences Oxford Brookes University Oxford, UK OX3 0BP
Just a reminder that TIFF is the officially recognized format by the Microscopy Society. Basic rule of thumb: save the original in TIFF then save a copy in JPEG for easier email communication etc.
The big problem with JPEG used to be degradation of the image on resizing, etc. I don't know if this is still the case with their new algorithms.
Hope this was helpful, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through 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 07:34 AM 5/31/2006, nizets2-at-yahoo.com wrote:
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==============================Original Headers============================== 13, 17 -- From bfoster-at-mme1.com Wed May 31 10:45:30 2006 13, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 13, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4VFjT9p014463 13, 17 -- for {microscopy-at-microscopy.com} ; Wed, 31 May 2006 10:45:30 -0500 13, 17 -- Received: (qmail 26875 invoked by uid 2020); 31 May 2006 11:02:47 -0500 13, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 13, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 31 May 2006 11:02:47 -0500 13, 17 -- Message-Id: {7.0.1.0.0.20060531104349.01ffc120-at-mme1.com} 13, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 17 -- Date: Wed, 31 May 2006 10:45:37 -0500 13, 17 -- To: nizets2-at-yahoo.com, microscopy Listserver {microscopy-at-microscopy.com} 13, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 13, 17 -- Subject: Re: [Microscopy] JPG and TIFF formats 13, 17 -- In-Reply-To: {200605311234.k4VCYvie019023-at-ns.microscopy.com} 13, 17 -- References: {200605311234.k4VCYvie019023-at-ns.microscopy.com} 13, 17 -- Mime-Version: 1.0 13, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Good evening, dear Fabio, in addition to the helpful message of Rick Powell on Silver Amplifying / Autometallographic localization of metals / Gorm Danscher papers etc. I would like to comment on your problem (since silver precipitation alone is more/less unspecific and will label a lot of metals in your specimen)....
Perhaps my message does not help you really, but if your } silver enhance kit { alone is not able to localize Cd qualitatively / quantitatively (and your Cd-concentration in the probe likely is very small) you could use also perhaps a precipitating method
} By means of crystalline precipitation as Cd (TH)2 [Cr (NH3)2 (CNS)4]2,1 0,1% of cadmium can be detected, without previous separation, in the presence of the metals of the ammonium sulphide group and the alkaline earths. Moreover, about 0,2% of cadmium can be detected by aid of a simple separation, in the presence of whatsoever metals of the hydrochloric acid and hydrogen sulphide groups. { source: Microchimica Acta Publisher: Springer Wien ISSN: 0026-3672 (Paper) 1436-5073 (Online) DOI: 10.1007/BF01471844 Issue: Volume 3, Number 4
Assuming that precipitation of Cd as CdS (with ammoniumsulfide, and -perhaps -using also KCN for masking copper) is not the method of your choice, I have had a look into PubMed and found the following reference: Biomarkers. 2002 Nov-Dec;7(6):491-500. Autometallography and metallothionein immunohistochemistry in hepatocytes of turbot (Scophthalmus maximus L.) after exposure to cadmium and depuration treatment. Amaral AF, Alvarado N, Marigomez I, Cunha R, Hylland K, Soto M. Section of Ecology, Department of Biology, University of the Azores, R Mae de Deus, 9500 Ponta Delgada, Sao Miguel, Azores, Portugal. Abstract: In this study, autometallography and immunohistochemistry were used to localize and quantify cadmium and metallothionein (MT) levels, respectively, in cellular compartments of turbot liver on exposure to cadmium for 7 days and further depuration treatment for 14 days. Metals weakly bound to proteins (i.e. MTs) in hepatocyte lysosomes were visualized as black silver deposits (BSDs) using a light microscope. With the aid of a newly developed immunohistochemical procedure, MTs were localized and semi-quantified in both the cytosolic and the lysosomal compartments of hepatocytes. The BSD extent in the lysosomes of hepatocytes increased significantly as a result of cadmium exposure. This response was evidenced after 1 h. Further, a progressive increase in the volume density of BSDs occurred up to the seventh day. Total MT immunohistochemical levels increased at a lower rate, starting after 1 day of cadmium exposure. BSD extent values recovered after depuration, whilst MT levels remain unchanged. It is possible that the detoxification rate of metals via lysosomes was diminished, whilst MT levels remained unchanged, at least after 14 days of depuration. It can be concluded that autometallography and MT immunohistochemistry are good tools for clarifying metal and metal-MT trafficking routes in hepatocytes, and also that BSD extent and MT immunohistochemical levels in the lysosomes and cytosol of fish hepatocytes can be considered to be useful biomarkers of metal exposure.
Unfortunately this is an article published in a Taylor and Francis Journal, so I was not able to retrieve the article as pdf.......but you can find the Abstract and Journal reference at: http://taylorandfrancis.metapress.com/link.asp?id=efeg34kqvvyn0vtg
The search in Pubmed for "cadmium localiz" resulted only with 17 hits.
Also I have found (for: "cadmium" and "EM" only 14 results) at: http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCP-4HHGNPH-1 &_coverDate=06%2F30%2F2006&_alid=408694459&_rdoc=1&_fmt=&_orig=search&_q d=1&_cdi=5176&_sort=d&view=c&_acct=C000057295&_version=1&_urlVersion=0&_ userid=2450241&md5=5a4679d3894972bb9ba28a4ab1f88de4
an article dealing with Cd-localization perhaps), also there only the abstract is available.
When using: } "cadmium" and "TEM" { as search phrase, I found a paper, entitled } Bioaccumulation and localization of exogenous cadmium in a teleost by electron microscopy (TEM) and its specific quantitation by electron probe X-ray microanalysis (EPMA) {. Bioorg Med Chem. 2000 Mar;8(3):475-82. by Tayal AK, Kaur I, Mathur RP.
the abstract of which tells us: } A cadmium bioconcentration study was carried out in a fresh water teleost, Colisa fasciatus, to study the bioaccumulation kinetics and fate of exogenous cadmium (Cd) in biological tissues. Study shows that on exposure of the fish to a sublethal concentration of cadmium in test water, Cd uptake results in its bioconcentration in gills, liver and muscle tissues. To explore whether the accumulated Cd reaches the membranes or inside the cells, transmission electron microscopy (TEM) of the thin sections of tissues was done after histochemical localization of Cd in cells by modified SST method.
TEM studies of sections of gills, liver and muscle tissues showed the deposits of exogenous Cd (visualized as dense clouds) in biological cells. This suggests the presence of free or loosely bound Cd on the membranes and inside the cells, which in the presence of Na2S is converted into insoluble metal sulfides. Electron probe X-ray microanalysis (EPMA) studies confirmed the presence of Cd on the membrane surface as well as inside the cells of bioindicator organs suggesting involvement of membrane transport of exogenous Cd inside the cells and its deposition as loosely bound insoluble metal complexes.
SST-Method: excerpt out of the pdf (which I am able to send to you, if you allow): } } } The SST process employs a developer with a low pH, silver nitrate as silver ion donator and a protecting colloid, gum arabic. The developer involves the use of small amounts of hydroquinone as reduction molecules. The SST results in reduced silver ion magnfications at specfic sites with gum arabic reducing the autocatalytic activity in the developer itself and the catalytic activity of the zone between the developer and the surface of the section [6,8]. However, SST visualizes only a certain fraction of heavy metals represented by free or loosely bound metal that in the presence of Na2S is converted into insoluble metal sufdes. The metal that is strongly bound to organic/inorganic ligands may not be sulfidated and remains histochemically invisible. The visualization of the fraction of Cd in fish tissue presented herein support Danscher and Norgaard [18] in that SST could be used for the demonstration of trace amounts of certain heavy metals at ultra-structural level and that the metal sufides can be shown by the technique. In their investigation, George et al.,[9] using the precipitation reactions (sufide for heavy metals), reported precipitation of Cd as a sufide in bivalve tissues. They also reported that precipitation reactions result in a better localization with a loss of 14% as compared to .....
Also, with this search phrase, you will discover some older references, like } Cellular alterations in collembolan midgut cells as a marker of heavy metal exposure: ultrastructure and intracellular metal distribution. { Sci Total Environ. 1996 Mar 29;181(3):187-200. by Pawert M, Triebskorn R, Graff S, Berkus M, Schulz J, Kohler HR. (which you can find at: http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V78-3VWF92S-1 P&_coverDate=03%2F29%2F1996&_alid=408705808&_rdoc=1&_fmt=&_orig=search&_ qd=1&_cdi=5836&_sort=d&view=c&_acct=C000057295&_version=1&_urlVersion=0& _userid=2450241&md5=77742f5cebfe3a73991ef90c3973adff )
the .pdf of which I also could send to you (please send } allowance { mail)
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Is it possible to use a immunogold purpose silver enhancement kit for revealing other heavy metals, such as cadmium, on paraffin or resin sections?
Best wishes
Fabio
Dr. Fabio D'Amico Dept. Biomedical Sciences University of Catania Via Androne 87/a I-95124 Catania ITALY tel/fax +39 095312017 email: f.damico-at-unict.it
==============================Original Headers============================== 7, 20 -- From fab-at-tariffenet.it Wed May 31 04:16:44 2006 7, 20 -- Received: from mbox.unict.it (mbox.unict.it [151.97.1.6]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4V9GiDq023000 7, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 31 May 2006 04:16:44 -0500 7, 20 -- Received: from Fabio.tariffenet.it (ssanfilippo.ipg.unict.it [151.97.169.115]) 7, 20 -- by mbox.unict.it (8.13.6/8.13.4) with ESMTP id k4V9GcWx017993 7, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 31 May 2006 11:16:38 +0200 7, 20 -- Message-Id: {7.0.1.0.0.20060531111607.01934818-at-unict.it} 7, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 7, 20 -- Date: Wed, 31 May 2006 11:16:37 +0200 7, 20 -- To: Microscopy-at-Microscopy.Com 7, 20 -- From: FD {fab-at-tariffenet.it} 7, 20 -- Subject: Silver enhancement kit 7, 20 -- Mime-Version: 1.0 7, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 7, 20 -- X-UniCT-MailScanner-Information: Please contact the ISP for more information 7, 20 -- X-UniCT-MailScanner: Found to be clean 7, 20 -- X-UniCT-MailScanner-SpamCheck: non spam, SpamAssassin (punteggio=-1.8, 7, 20 -- necessario 5, autolearn=not spam, ALL_TRUSTED -1.80) 7, 20 -- X-MailScanner-From: fab-at-tariffenet.it ==============================End of - Headers==============================
==============================Original Headers============================== 39, 28 -- From W.Muss-at-salk.at Wed May 31 11:28:09 2006 39, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 39, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k4VGS83A025354 39, 28 -- for {Microscopy-at-Microscopy.Com} ; Wed, 31 May 2006 11:28:08 -0500 39, 28 -- Received: from localhost (localhost [127.0.0.1]) 39, 28 -- by hermes.lks.at (Postfix) with ESMTP id DCC3E5A9044; 39, 28 -- Wed, 31 May 2006 18:28:05 +0200 (CEST) 39, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 39, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 39, 28 -- with ESMTP id 68448-04; Wed, 31 May 2006 18:28:05 +0200 (CEST) 39, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 39, 28 -- by hermes.lks.at (Postfix) with SMTP id 495455A900A; 39, 28 -- Wed, 31 May 2006 18:28:05 +0200 (CEST) 39, 28 -- Received: by localhost with Microsoft MAPI; Wed, 31 May 2006 18:28:02 +0200 39, 28 -- Message-ID: {01C684DF.F38CC420.W.Muss-at-salk.at} 39, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 39, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 39, 28 -- To: "'fab-at-tariffenet.it'" {fab-at-tariffenet.it} 39, 28 -- Cc: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 39, 28 -- Subject: [Microscopy]Re: (LONG) Silver enhancement kit 39, 28 -- Date: Wed, 31 May 2006 18:27:58 +0200 39, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 39, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 39, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 39, 28 -- MIME-Version: 1.0 39, 28 -- Content-Type: text/plain; charset="us-ascii" 39, 28 -- Content-Transfer-Encoding: 7bit 39, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
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A user wants to remove the data bar from a stored image. Can anyone remind me whether this is possible apart from cropping?
Dave
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==============================Original Headers============================== 5, 31 -- From David.Patton-at-uwe.ac.uk Wed May 31 11:42:42 2006 5, 31 -- Received: from mailapp01.uwe.ac.uk (mailapp01.uwe.ac.uk [164.11.132.61]) 5, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4VGgfpx003093 5, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 31 May 2006 11:42:41 -0500 5, 31 -- Received: from (164.11.132.62) by mailapp01.uwe.ac.uk via smtp 5, 31 -- id 3098_11e6e1a2_f0c5_11da_8ef2_0002b3c946e4; 5, 31 -- Wed, 31 May 2006 17:46:56 +0100 5, 31 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 5, 31 -- (egen-uwe01.campus.ads.uwe.ac.uk [164.11.249.121]) 5, 31 -- by mta02.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 5, 31 -- 2005)) with ESMTP id {0J05007CA2F5J2-at-mta02.uwe.ac.uk} for 5, 31 -- Microscopy-at-microscopy.com; Wed, 31 May 2006 17:42:41 +0100 (BST) 5, 31 -- Date: Wed, 31 May 2006 17:42:40 +0100 5, 31 -- From: David Patton {David.Patton-at-uwe.ac.uk} 5, 31 -- Subject: Question for FEI XL30 Users 5, 31 -- To: Microscopy-at-microscopy.com 5, 31 -- Message-id: {F247F674896BE243AD8263C5280E2BDB024A665C-at-egen-uwe01} 5, 31 -- MIME-version: 1.0 5, 31 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.6944.0 5, 31 -- Content-type: text/plain; charset=us-ascii 5, 31 -- Content-class: urn:content-classes:message 5, 31 -- Thread-topic: Question for FEI XL30 Users 5, 31 -- Thread-index: AcaE0Tta1uyunzIuSLuoDwJ+qDEQjg== 5, 31 -- X-MS-Has-Attach: 5, 31 -- X-MS-TNEF-Correlator: 5, 31 -- X-NAI-Spam-Score: 0 5, 31 -- X-NAI-Spam-Rules: 0 Rules triggered 5, 31 -- X-NAIMIME-Disclaimer: 1 5, 31 -- X-NAIMIME-Modified: 1 5, 31 -- Content-Transfer-Encoding: 8bit 5, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k4VGgfpx003093 ==============================End of - Headers==============================
My recent response to ESEM detectors was subjective and incomplete. My errors have brought Daniel Phifer, the application specialist of FEI to my rescue. Please read below. Daniel, thank you.
Ann Fook Yang EM Unit/ Unite EM Bldg 20, AAFC/AAC 960 Carling Ave, Ottawa, Ontario Canada K1A 0C6 Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 yanga-at-agr.gc.ca
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada ============================================================================
AnnFook, I was forwarded your list server response and wanted to give you some info to better respond.
For your samples which have little to no atomic contrast, there is likely no need for the BSE except when looking for gold labels, etc. This is usually on dried prepared biological samples. With the majority of your samples at Agri-Food Canada, the BSE and SE images may be similar as the atomic composition is very similar.
The query could be from a customer who has lots of atomic differences and thus may need a BSE detector to show the atomic differences. Maybe not, but this should be qualified to give the best response.
FEI has two detectors for picking up BSE signal. The solid state BSE crystal can be used in high and low vacuum but not in ESEM (as it requires the gas to amplify the signal. As the pressure is increased above 2-3 Torr in the chamber, the BSE crystal will get less of the signal reaching the detector as the BSE collide with the gas and scatter. For this situation, there is an optional Gaseous Backscattered Detector (GBSD) which can be used in ESEM mode (4-7Torr optimum). The GBSD will give atomic contrast (even in fully hydrated samples) using a patented detection mechanism. If BSE is desired, these two detectors are the best options. The GSED (Gaseous Secondary Electron Detector) only picks up secondary electrons which yield surface information, not composition.
It is my impression from the question that the terminology is getting confused in translation. Hopefully this information will allow you to clarify your response. Please feel free to paraphrase or plagiarize my words and make them your own if you like. I just want you to have the information. If you have questions about the technology and would like me to help you answer a query, please do not hesitate to contact me directly.
Hope all is well in Ottawa.
Daniel
Daniel Phifer
FEI Company
-----Original Message----- X-from: Yang, Ann-Fook Sent: Tuesday, May 30, 2006 11:38 AM To: Yang, Ann-Fook
You cannot use GSED in high vac; it is only for low vac. I have a XL30 ESEM and never use the backscatter detector (BSD) since installation six years ago for morphological study, except using it to compare with GSED at the very beginning. A GSED produces images a lot better than a BSD. If I were you, the pick would have been a GSED. If you need a BSD for another reason, then that is another matter to consider. As for software, you don't have to worry; just put in a new GSED and off you go. I have checked with my service engineer.
Ann Fook Yang EM Unit/ Unite EM Bldg 20, AAFC/AAC 960 Carling Ave, Ottawa,Ontario Canada K1A 0C6 Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 yanga-at-agr.gc.ca
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: Amr_elsirafy-at-hotmail.com [mailto:Amr_elsirafy-at-hotmail.com] Sent: Tuesday, May 30, 2006 9:21 AM To: Yang, Ann-Fook
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Email: Amr_elsirafy-at-hotmail.com Name: afaf Ibrahim Mahdi
Organization: Cairo university
Title-Subject: [Filtered] ESEM-detectors
Question: Hello I' d like to know is GSED is used in ESEM with tungesten filament and if it can be used in both low and high vacuum mode because the BSD in my XL30 TMP instrument is damaged and I want to know iif it is best to buy back scatter detectors or GSED and is it need a new software.
Another informative listserver for FEI/Philips XL30 ESEM's is maintained by Dr. Cameron Begg at Ohio State University. See instructions below. I hope this paste is still valid.
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} Just a reminder that TIFF is the officially recognized format } by the Microscopy Society.
More exactly ... It is the ^uncompressed^ TIFF that is the MSA standard format ... Presumably because the bitmap is intact, while compression algorithms come and go.
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} {http://www.esd.mun.ca/epma/} Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
} } At 07:34 AM 5/31/2006, 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 } } ------------------------------------------------------------- } ---------- } } ----- } } } } Hello, } } } } The problem of Eric brings a new question to me: } } The TEM pictures may be saved in JPG or TIFF formats. } } I know that TIFF format keeps all the information without } compression. } } But does JPG without compression (quality 12) deteriorates } the quality } } of the picture? } } I thought that JPG without compression was as good as TIFF. } } } } Stephane } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam protection around } } http://mail.yahoo.com } } } } ==============================Original } } Headers============================== } } 4, 18 -- From nizets2-at-yahoo.com Wed May 31 07:31:48 2006 4, 18 -- } } Received: from web37414.mail.mud.yahoo.com } (web37414.mail.mud.yahoo.com [209.191.87.67]) } } 4, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id k4VCVlda014756 } } 4, 18 -- for {microscopy-at-microscopy.com} ; Wed, 31 May } 2006 07:31:47 -0500 } } 4, 18 -- Received: (qmail 79599 invoked by uid 60001); 31 May 2006 } } 12:31:47 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; } q=dns; c=nofws; } } 4, 18 -- s=s1024; d=yahoo.com; } } 4, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Conten } t-Type:Content-Transfer-Encoding; } } 4, 18 -- } b=OR+0f0jaY3NylTkKHJTgGTfxY3WS4kLrGgRmUZ+3VSncfFVMPDvyj1Znnxyo } cY+3mH9BSMuQQ9vtla/UCLW81Yy8dCdwp9bsESZ8PVDCoKZF9jVntYwHDeYB3S sLEYlQfixGwGwS4jBQxGeBRgpLub2T5Zxfcra0ICLmIxsbvmE= ; } } 4, 18 -- Message-ID: } } {20060531123147.79597.qmail-at-web37414.mail.mud.yahoo.com} } } 4, 18 -- Received: from [80.122.101.102] by } web37414.mail.mud.yahoo.com } } via HTTP; Wed, 31 May 2006 05:31:47 PDT 4, 18 -- Date: Wed, } 31 May 2006 } } 05:31:47 -0700 (PDT) 4, 18 -- From: Stephane Nizet } {nizets2-at-yahoo.com} } } 4, 18 -- Subject: JPG and TIFF formats 4, 18 -- To: } } microscopy-at-microscopy.com 4, 18 -- MIME-Version: 1.0 4, 18 -- } } Content-Type: text/plain; charset=iso-8859-1 4, 18 -- } } Content-Transfer-Encoding: 8bit } ==============================End of - } } Headers============================== } } } ==============================Original } Headers============================== } 13, 17 -- From bfoster-at-mme1.com Wed May 31 10:45:30 2006 13, } 17 -- Received: from 5starpro.com (enterprise.5starpro.com } [207.44.136.95]) } 13, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id k4VFjT9p014463 } 13, 17 -- for {microscopy-at-microscopy.com} ; Wed, 31 May } 2006 10:45:30 -0500 } 13, 17 -- Received: (qmail 26875 invoked by uid 2020); 31 May } 2006 11:02:47 -0500 13, 17 -- Received: from } host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) } 13, 17 -- by enterprise.5starpro.com with } (DHE-RSA-AES256-SHA encrypted) SMTP; 31 May 2006 11:02:47 -0500 } 13, 17 -- Message-Id: {7.0.1.0.0.20060531104349.01ffc120-at-mme1.com} } 13, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } 13, 17 -- Date: Wed, 31 May 2006 10:45:37 -0500 13, 17 -- To: } nizets2-at-yahoo.com, microscopy Listserver {microscopy-at-microscopy.com} } 13, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 13, 17 -- } Subject: Re: [Microscopy] JPG and TIFF formats 13, 17 -- } In-Reply-To: {200605311234.k4VCYvie019023-at-ns.microscopy.com} } 13, 17 -- References: {200605311234.k4VCYvie019023-at-ns.microscopy.com} } 13, 17 -- Mime-Version: 1.0 } 13, 17 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== }
==============================Original Headers============================== 7, 21 -- From Michael-at-Shaffer.net Wed May 31 13:50:22 2006 7, 21 -- Received: from ws6-5.us4.outblaze.com (ws6-5.us4.outblaze.com [205.158.62.152]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4VIoL4l003540 7, 21 -- for {Microscopy-at-microscopy.com} ; Wed, 31 May 2006 13:50:21 -0500 7, 21 -- Received: (qmail 31139 invoked from network); 31 May 2006 18:50:20 -0000 7, 21 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 7, 21 -- by ws6-5.us4.outblaze.com with SMTP; 31 May 2006 18:50:20 -0000 7, 21 -- From: "michael shaffer" {michael-at-Shaffer.net} 7, 21 -- To: {bfoster-at-mme1.com} 7, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 21 -- Subject: RE: [Microscopy] Re: JPG and TIFF formats 7, 21 -- Date: Wed, 31 May 2006 16:20:19 -0230 7, 21 -- Message-ID: {003e01c684e3$113ef8a0$8d829986-at-roamingwolf} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 7, 21 -- Thread-Index: AcaEyWHBfsgDrt3xTX2+IpK0bgAjkQAGSztw 7, 21 -- In-Reply-To: {200605311546.k4VFkQmv015265-at-ns.microscopy.com} ==============================End of - Headers==============================
Sorry, I was incommunicado for a few days and just saw this posting.
The resolution information is not stored in the tags mentioned below for the following reason: Many word processing and page layout programs use that tag to print something at the "right" size (for example MS Word). If you enter the calibration value in this field, the software will try to print the image with a width of a few microns, resulting in a single dot on the paper. We ran into this behavior a few years ago and were forced to store the calibration values in a private tag. If you want to know what the tag is, let me know, I can find out for you.
ananlySIS does not automatically convert images to 8 bit, unless instructed to do so. One option is, as already mentioned, the possibility to automatically convert to 8-bit when saving the image. Of course, if that is selected, the software will do that.
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: eric.leroy-at-glvt-cnrs.fr [mailto:eric.leroy-at-glvt-cnrs.fr] Sent: Wednesday, May 31, 2006 3:45 To: Mike Bode
David,
Thank you for the information. I downloaded the software you told me but unfortunately, the information written int Xresolution tag is allways 200 and the ResolutionUnit is 0. So it seems that the saptial calibration is stored elsewhere.
Best regards
Eric
Le 30/05/06 17:58, « David Vowles » {djv23-at-cam.ac.uk} a écrit :
} Eric, } } The TIFF standard allows for spatial resolution data to be saved in the } fields XResolution (no of pixels per measurement unit in X dir) and } YResolution (no of pixels per measurement unit in Y dir). The measurement } unit data is contained in the ResolutionUnit field (usually inch or cm). } From these you should be able to calculate the pixel size, magn or image } width. There are a number of free TIFF tag viewers available, for example, } http://www.awaresystems.be/imaging/tiff/astifftagviewer.html, or just } Google 'tiff tag viewer' . } Hope this helps. } } David Vowles } Electron Microscope Unit } Dept of Materials Science and Metallurgy } University of Cambridge } Pembroke St Cambridge } UK CB2 3QZ } Tel: +44 (0)1223 334325 } Fax: +44 (0)1223 334567 } Email: djv23-at-cam.ac.uk } } } } } Name: Eric Leroy } } } } Organization: CNRS - LCMTR } } } } Title-Subject: [Filtered] AnalySIS TIFF images } } } } Question: Hi, } } } } We have a SIS Megaview III camera attached to our Tecnai and we use } } AnalySIS to record the images. Since we only have a full version of } } AnalySIS installed on the microscope's PC we usually treat ours images } } with ImageJ. ImageJ can perfectly read the TIFF images but is unable to } } read the spatial calibration of the image. Do you know how this spatial } } calibration is written in the image ? } } } } Thanks } } } } Eric } } } } --------------------------------------------------------------------------- } } } } ==============================Original Headers============================== } } 9, 12 -- From zaluzec-at-microscopy.com Tue May 30 08:08:56 2006 } } 9, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } } 9, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } } id k4UD8s0s005952 } } 9, 12 -- for {microscopy-at-microscopy.com} ; Tue, 30 May 2006 08:08:56 } } -0500 } } 9, 12 -- Mime-Version: 1.0 } } 9, 12 -- X-Sender: (Unverified) } } 9, 12 -- Message-Id: {p06110404c0a1f45139b1-at-[206.69.208.22]} } } 9, 12 -- Date: Tue, 30 May 2006 08:08:49 -0500 } } 9, 12 -- To: microscopy-at-microscopy.com } } 9, 12 -- From: eric.leroy-at-glvt-cnrs.fr (by way of MicroscopyListserver) } } 9, 12 -- Subject: viaWWW: AnalySIS TIFF images } } 9, 12 -- Content-Type: text/plain; charset="us-ascii" } } ==============================End of - Headers============================== }
Eric LEROY Dr. Laboratoire de Chimie Metallurgique des Terres Rares UPR 209 - CNRS Groupe des Laboratoires de Thiais 2-8, rue Henri Dunant 94320 THIAIS cedex
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Email: afaf_mahdi-at-hotmail.com Name: Afaf Ibrahim Mahdi
Organization: Cairo Univ
Title-Subject: [Filtered] ESEM detectors
Question: Hello all I have already recieved answers from some scientists for my question about GSED and it was valuable. I'am a mieralogist and I used ESEM XL30 TMP in examining geological samples and also biological samples. I used BSD fo about 90% of my work. For about 6 years I used BSD in both high vacuum and low vacuum modes by converting Key no 4 and I used only carbon sticker tofix samples. it gives satisfied EDX analyses but images some time satisfactory and sometimes it is very bad. The magnification dont exceed 8000X. Now the back scatter detector is damage and I want to buy one suitable for both modes because I don' have enough moner to buy 2 detectors. Is there one detector can used in both modes taking into account that i still have GSD or it is better to buy BSD. In the last monty I noticed that the two pumps are begging to heat severly, could be the detector is the reason. thank you
} Just a reminder that TIFF is the officially recognized format by the Microscopy Society. Basic rule of thumb: save the original in } TIFF then save a copy in JPEG for easier email communication etc. } } The big problem with JPEG used to be degradation of the image on resizing, etc. I don't know if this is still the case with their new } algorithms.
JPEG2000 is no more lossless than the original JPEG - only the method of compression has changed.
Also - I warn people to beware of TIFFs from unknown programs. The TIFF 6 specification does allow for storing JPEG compressed data in the TIFF wrapper - there is a flag that indicates the compression level.
I need to make samples disbersed on carbon-coated grids for collecting some EELS standards of various oxides in powder form. I have Maganese, Titiatium, and Ruthenium oxides in various states. Anyone know a good prep that will evenly disburse a thin layer onto a carbon-coated grid? My first thought would be to mix with a solvent and put a drop on a grid, but I figured someone here has probably already done this.
Thanks in advance, Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 6, 26 -- From lkrupp-at-us.ibm.com Thu Jun 1 10:39:52 2006 6, 26 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51Fdq5O008963 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 10:39:52 -0500 6, 26 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 6, 26 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k51FdZKg024920 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 11:39:35 -0400 6, 26 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 6, 26 -- by d01relay02.pok.ibm.com (8.12.10/NCO/VER6.8) with ESMTP id k51FdKSm103888 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 11:39:35 -0400 6, 26 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 6, 26 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k51FdKAl024003 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 11:39:20 -0400 6, 26 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 6, 26 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k51FdKjj023990 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 11:39:20 -0400 6, 26 -- Subject: Sample prep for disbersing powder samples 6, 26 -- To: microscopy-at-microscopy.com 6, 26 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 6, 26 -- Message-ID: {OFF13B2E87.F51B825E-ON85257180.005561A1-88257180.0055CAE6-at-us.ibm.com} 6, 26 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 6, 26 -- Date: Thu, 1 Jun 2006 08:38:56 -0700 6, 26 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 6, 26 -- 06/01/2006 11:39:19 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
On Jun 1, 2006, at 8:40 AM, lkrupp-at-us.ibm.com wrote:
} I need to make samples disbersed on carbon-coated grids for collecting } some } EELS standards of various oxides in powder form. I have Maganese, } Titiatium, and Ruthenium oxides in various states. Anyone know a good } prep } that will evenly disburse a thin layer onto a carbon-coated grid? My } first } thought would be to mix with a solvent and put a drop on a grid, but I } figured someone here has probably already done this. } Dear Leslie, My choice would be your first thought--it worked very well for river-bottom sediment. I would try various dilutions to find the one that gives the desired distribution of particles, and I would glow-discharge the grids. If using water leads to aggregation, try an organic solvent, and if the particles still have a tendency to aggregate, try putting the grid with the drop of solvent in an oven--faster evaporation might overcome that tendency. 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 Jun 1 12:10:36 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 k51HAawk020886 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 12:10:36 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id BEB94366A6 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 10:10:35 -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 3A54510AE0C 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 10:10:30 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606011540.k51Fe4m9009135-at-ns.microscopy.com} 4, 22 -- References: {200606011540.k51Fe4m9009135-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {a6cd43e8d0c28b6d5fc583cdda84c003-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Sample prep for disbersing powder samples 4, 22 -- Date: Thu, 1 Jun 2006 10:21:39 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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Email: Henrik.Kaker-at-guest.arnes.si Name: Henrik Kaker
Organization: SEM-EDS Lab, Metal Ravne, Slovenia
Title-Subject: [Filtered] Equipment for phase extraction from steels
Question: Dear All,
I need to choose equipment for electrolytical phase extraction from steels (carbides, nitrides, etc.) and I need any advice about equipment and procedures for electrolytical phase extraction. We need this equipment for sample preparation for XRD. Thanks in advance.
A colleague has to serial section samples embedded in Araldite. He is trimming the block using a diamond trimming tool and is trimming to a very small block face. The problem is that he is not getting ribbons formed when he sections. Does anyone have a trick to get the sections to stick together and form ribbons? Thanks in advance.
Bob Temkin
Advanced Bioimaging Centre Mount Sinai Hospital Pathology and Laboratory Medicine
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Bill, We too have used the method you've described, but I am not familiar with "glow discharge the grids". What is tis and how does it work?
Thanks, Lou Ross
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==============================Original Headers============================== 6, 19 -- From RossLM-at-missouri.edu Thu Jun 1 13:44:31 2006 6, 19 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51IiUUf020636 6, 19 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 13:44:31 -0500 6, 19 -- Received: from um-exproto9.um.umsystem.edu ([207.160.151.49]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 6, 19 -- Received: from [128.206.78.127] ([128.206.78.39]) by um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft SMTPSVC(6.0.3790.1830); 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 6, 19 -- Mime-Version: 1.0 6, 19 -- X-Sender: RossLM-at-pop.missouri.edu 6, 19 -- Message-Id: {p05200f2fc0a4e55028d2-at-[128.206.78.127]} 6, 19 -- In-Reply-To: {200606011711.k51HBix4023116-at-ns.microscopy.com} 6, 19 -- References: {200606011711.k51HBix4023116-at-ns.microscopy.com} 6, 19 -- Date: Thu, 1 Jun 2006 13:44:29 -0500 6, 19 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 19 -- From: Lou Ross {RossLM-at-missouri.edu} 6, 19 -- Subject: [Microscopy] Re: Sample prep for disbersing powder samples 6, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 19 -- X-OriginalArrivalTime: 01 Jun 2006 18:44:30.0740 (UTC) FILETIME=[6AFB6540:01C685AB] ==============================End of - Headers==============================
I would like to be short....."glow discharging" is a kind of creating a kind of "air plasma" in order to get formvar-coated grids stickier (= more hydrophilic, discharging of surface charge).
This process necessarily /usually is done in a sputter coater or "plasma etcher" or the like (must have a recipient and vacuum pump(s) rotary, and - if (perhaps) you need higher vac, diffusion oil or tubomolecular pump...) as well as an inbuilt ionizing source.
Often such a technique is used for getting formvar-coated grids (used for negative staining procedure on virus particles) again hydrophilic as well as stickier for the adsorption of virus particles to the formvar film. I enclose here - FY convenience - a thread on that issue published in Microscopy Today (and therefore was a theme via the MSA Listserver in 2005.........
NetNotes aus MICROSCOPY TODAY Vol. 13, No 3, May 2005, p. 64-65
Formvar Grids (de-charging by glow discharging and other methods) ---------------- Q: We have been using Formvar/carbon coated grids for spreading magnetic nano-particles in water and for negative staining. But as you know, an aqueous solution does not spread well on grids because of the charge characteristics on the film surface. The only way I know of for making the film surface more hydrophilic is to do a glow discharge in a sputter coater, but we do not have such a device. I heard treating coated grids with ethanol vapor works, but not for me. Does anyone have any other tricks or suggestions? Hong Yi {hyi-at-emory.edu} 20 Feb 2005 ----------- Answers: Do you have a vacuum evaporator? It's simple to rig it for glow discharge with an inexpensive Tesla coil. A plastic vacuum desiccator, the same Tesla coil, and a rough vacuum source will do the job also. Caroline Schooley {schooley-at-mcn.org} 21 Feb 2005 ------------ I don't think that a splitter coater, old or new, is going to solve your problem. I have not heard of ethanol vapors making a carbon grid more hydrophilic. Carbon coated grids lose their hydrophilic nature as they age and they become more hydrophobic. The process can be "reversed" by (a) exposure to an RF "air" plasma in a small plasma etcher (effect will last 60-90 days) or (b) a thin evaporation of VictawetO onto the grids. Our own studies would suggest that Victawet can keep the grids highly hydrophilic essentially forever (e.g. more than one year). Just remember that it is a phosphate based surfactant so if you are doing elemental analysis work, you might not want to have P showing up in your data. But if you have an ordinary vacuum evaporator and tungsten baskets, and don't have a plasma etcher, you can solve your problem with Victawet. The best bet for having carbon coated grids with the greatest hydrophilic characteristics is to make or purchase your carbon coated grids always "fresh': If the grids are purchased, and their age is uncertain, contact the manufacturer of the carbon coated grids, give them the lot number and then you will know. Charles A. Garber {cgarber-at-2spi.com} 21 Feb 2005 ---------- Another method occasionally used to make C-coated grids hydrophilic is to expose them for some minutes to the direct illumination of a UV lamp. This is done at normal atmospheric pressure, so it needs no vacuum technology. James Chalcroft {jchalcro-at-neuro.mpg.de} 21 Feb 2005 ---------- I personally don't like glow discharge at all: it's very difficult to reproduce. It depends on the equipment and there is no way to control the "amount" of discharge. I use 0.5-1 % poly-lysine from any of the EM suppliers (don't try to make the solution yourself since there is some trick required). Place the EM grid on a 10 ?l poly-lysine drop for 5-10 min, wash on a few drops of deionized water, air dry - good for at least a month. Alcian Blue works in the similar way with a similar result. Of course, these methods will only work for positively charged molecules. Sergey Ryazantsev {sryazant-at-ucla.edu} 22 Feb 2005
hope this answers your question... best regards
Wolfgang MUSS Salzburg, Austria =============================} } { {======================== ---------- Von: RossLM-at-missouri.edu[SMTP:RossLM-at-missouri.edu] Antwort an: RossLM-at-missouri.edu Gesendet: Donnerstag, 01. Juni 2006 20:50 An: W.Muss-at-salk.at Betreff: [Microscopy] Re: Sample prep for disbersing powder samples
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Thanks, Lou Ross
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Bill, We too have used the method you've described, but I am not familiar with "glow discharge the grids". What is tis and how does it work?
Thanks, Lou Ross
--------------------------
Hello Lou,
Essentially, glow discharge is a procedure that you can carry out on a standard vacuum evaporator that is equipped with a high voltage AC input. Unfortunately, not many vac evaporators have this accessory. In operation, specimens are loaded into the chamber and one end of about 8-12 inches of high purity aluminum wire (3-5 mm) is plugged into the HV feedthrough and a large loop is formed to surround the grids. The chamber is pumped down using only the rotary pump until you get approximately 50-100 millitorr vacuum. At this point, you turn up the AC high voltage (using a variAC) and you will generate a plasma similar to what one sees in a sputter coater. The plasma is thought to both clean the grid surface as well as imparting a charge that renders the grid surface (of carbon-coated grids) hydrophilic. The effect lasts about a day--less if in a high humidity environment.
We have been able to approach this technique by placing the grids inside a sputter coater and shielding them from direct line of sight with the target (to prevent coating with metal). The plasma generated will impart a hydrophobic character to the grids. But you may also get metal deposition, so do some trial runs first.
I have been depositing a lot of nanocrystals as you describe by directly depositing them on carbon or silicon substrates. The powders are suspended in acetone, shaken and (after allowing the large particles to settle) a small volume taken up in a micropitettor (about 10-20 microliters). This is then dropped directly onto the grid surface from a distance of 10 or so mm.
I can take a picture of the glow discharge setup if you like, Lou.
JB
} } } } On Jun 1, 2006, at 8:40 AM, lkrupp-at-us.ibm.com wrote: } } } } } I need to make samples disbersed on carbon-coated grids for collecting } } } some } } } EELS standards of various oxides in powder form. I have Maganese, } } } Titiatium, and Ruthenium oxides in various states. Anyone know a good } } } prep } } } that will evenly disburse a thin layer onto a carbon-coated grid? My } } } first } } } thought would be to mix with a solvent and put a drop on a grid, but I } } } figured someone here has probably already done this. } } } } } Dear Leslie, } } My choice would be your first thought--it worked very well for } } river-bottom sediment. I would try various dilutions to find the one } } that gives the desired distribution of particles, and I would } } glow-discharge the grids. If using water leads to aggregation, try an } } organic solvent, and if the particles still have a tendency to } } aggregate, try putting the grid with the drop of solvent in an } } oven--faster evaporation might overcome that tendency. } } 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 Jun 1 12:10:36 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 k51HAawk020886 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 } } 12:10:36 -0500 } } 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) } } 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id BEB94366A6 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 } } 10:10:35 -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 3A54510AE0C } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 } } 10:10:30 -0700 (PDT) } } 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) } } 4, 22 -- In-Reply-To: {200606011540.k51Fe4m9009135-at-ns.microscopy.com} } } 4, 22 -- References: {200606011540.k51Fe4m9009135-at-ns.microscopy.com} } } 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } } 4, 22 -- Message-Id: {a6cd43e8d0c28b6d5fc583cdda84c003-at-caltech.edu} } } 4, 22 -- Content-Transfer-Encoding: 7bit } } 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} } } 4, 22 -- Subject: Re: [Microscopy] Sample prep for disbersing powder samples } } 4, 22 -- Date: Thu, 1 Jun 2006 10:21:39 -0700 } } 4, 22 -- To: microscopy-at-msa.microscopy.com } } 4, 22 -- X-Mailer: Apple Mail (2.624) } } 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 } } ==============================End of - Headers============================== } } } -- } 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============================== } 6, 19 -- From RossLM-at-missouri.edu Thu Jun 1 13:44:31 2006 } 6, 19 -- Received: from um-nsmtpout1.um.umsystem.edu } (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) } 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k51IiUUf020636 } 6, 19 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 13:44:31 -0500 } 6, 19 -- Received: from um-exproto9.um.umsystem.edu } ([207.160.151.49]) by um-nsmtpout1.um.umsystem.edu with Microsoft } SMTPSVC(6.0.3790.1830); } 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 } 6, 19 -- Received: from [128.206.78.127] ([128.206.78.39]) by } um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft } SMTPSVC(6.0.3790.1830); } 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 } 6, 19 -- Mime-Version: 1.0 } 6, 19 -- X-Sender: RossLM-at-pop.missouri.edu } 6, 19 -- Message-Id: {p05200f2fc0a4e55028d2-at-[128.206.78.127]} } 6, 19 -- In-Reply-To: {200606011711.k51HBix4023116-at-ns.microscopy.com} } 6, 19 -- References: {200606011711.k51HBix4023116-at-ns.microscopy.com} } 6, 19 -- Date: Thu, 1 Jun 2006 13:44:29 -0500 } 6, 19 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} } 6, 19 -- From: Lou Ross {RossLM-at-missouri.edu} } 6, 19 -- Subject: [Microscopy] Re: Sample prep for disbersing powder samples } 6, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 6, 19 -- X-OriginalArrivalTime: 01 Jun 2006 18:44:30.0740 (UTC) } FILETIME=[6AFB6540:01C685AB] } ==============================End of - Headers==============================
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Vendors who recondition LKB 7801 B knife makers please contact me. -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 1, 20 -- From oshel1pe-at-cmich.edu Thu Jun 1 14:54:26 2006 1, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 1, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51JsP7S011509 1, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 14:54:26 -0500 1, 20 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 1, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k51KTl4l023712 1, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 16:29:47 -0400 1, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 1, 20 -- Thu, 1 Jun 2006 15:54:25 -0400 1, 20 -- Mime-Version: 1.0 1, 20 -- Message-Id: {f0623090ac0a4f6398d5a-at-[141.209.160.132]} 1, 20 -- Date: Thu, 1 Jun 2006 15:54:24 -0400 1, 20 -- To: Microscopy-at-microscopy.com 1, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 1, 20 -- Subject: LKB knifemaker service 1, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 1, 20 -- X-OriginalArrivalTime: 01 Jun 2006 19:54:25.0358 (UTC) FILETIME=[2F2B4EE0:01C685B5] 1, 20 -- X-CanItPRO-Stream: default 1, 20 -- X-Spam-Score: -4 () L_EXCH_MF 1, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
} ### apologies for multiple postings } } } } } The Department of Sedimentology and Environmental Geology, } Geoscience Center of the University } of Goettingen, is offering a } } } post-doc position in Sedimentology / Sedimentary Petrology } } } beginning 1st October, 2006. The appointment is for one year with an } option for another six month. } Salary is based on the wage agreement of the German civil service } (75% BAT IIa, approx. 30.000 EUR p.a.). } } The Geoscience Center in Goettingen offers a wide range of } state-of-the-art analytical facilities and an excellent environment } for geological, sedimentological and geochemical research ( } http://www.gzg.uni-goettingen.de ). } We are seeking for a gifted and motivated scientist with a several } years perspective for our research team. } } The successful candidate has finished his/her excellent doctoral } thesis no longer than two years ago. Ongoing and future research } should focus on one of the major research topics of the department, } such as exhumation & erosion, weathering processes, sediment } geochemistry and/or } provenance analysis. During the one-year period offered here a } research proposal should be set up to continue research on soft money. } } Teaching duties comprise sedimentary petrology as well as } sedimentary facies and basin analysis at } various levels, as well as field trips. The University of Goettingen } is attempting to increase the } proportion of women on the scientific staff and strongly encourages } qualified women to apply. } Disabled candidates with equal qualification will be preferred. } } Applicants should send a letter of interest, curriculum vitae, a } brief statement of research interests, list of publications and } contact information for two referees, no later than June 30, 2006, } to Prof. Dr. Hilmar von Eynatten, Department of Sedimentology and } Environmental Geology, Geoscience Center Goettingen, } Goldschmidtstrasse 3, D-37077 Goettingen, Germany } (hilmar.von.eynatten-at-geo.uni-goettingen.de, } http://www.sediment.uni-goettingen.de ). }
==============================Original Headers============================== 4, 20 -- From tamas.mikes-at-geo.uni-goettingen.de Fri Jun 2 09:52:03 2006 4, 20 -- Received: from mailer.gwdg.de (mailer.gwdg.de [134.76.10.26]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k52Eq205001096 4, 20 -- for {microscopy-at-microscopy.com} ; Fri, 2 Jun 2006 09:52:03 -0500 4, 20 -- Received: from sedi8.gzg.geo.uni-goettingen.de ([134.76.77.171] helo=sedi8.geo.uni-goettingen.de) 4, 20 -- by mailer.gwdg.de with esmtp (Exim 4.60) 4, 20 -- (envelope-from {tamas.mikes-at-geo.uni-goettingen.de} ) 4, 20 -- id 1FmB0I-0006jz-7I 4, 20 -- for microscopy-at-microscopy.com; Fri, 02 Jun 2006 16:52:02 +0200 4, 20 -- Message-Id: {7.0.1.0.0.20060602165219.0227a738-at-geo.uni-goettingen.de} 4, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 4, 20 -- Date: Fri, 02 Jun 2006 16:53:12 +0200 4, 20 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 4, 20 -- From: Tamas Mikes {tamas.mikes-at-geo.uni-goettingen.de} 4, 20 -- Subject: post-doc position in sedimentary petrology available 4, 20 -- Mime-Version: 1.0 4, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 4, 20 -- X-Spam-Level: / 4, 20 -- X-Spam-Report: Content analysis: 0.0 points, 6.0 required 4, 20 -- X-Virus-Scanned: (clean) by exiscan+sophie ==============================End of - Headers==============================
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I have a SEM sample of bacteria in milk. Does anyone have any suggestions on how to reduce the milk debris/fat leaving the bacteria intact? I have centrifuged the sample, took from the formed pellet and deposited onto a millipore filter on a pump, buffer washed several times and followed a standard fixation protocol though HMDS. Bacteria of other samples (not in milk) looked great, but the milk sample left way too much debris behind.
Thanks in advance for any suggestions. Lynda Schneider University of Florida
It's that time again! We are in need for volunteers for the M&M meeting in Chicago.
Student volunteers will be paid through a voucher system. Other volunteers will receive benefits according to the hours provided. As soon as you are accepted as a volunteer, your name will be provided to meeting registration. All efforts will be made to place volunteers for sessions and booths that they are interested in.
For more information and a schedule of sessions/jobs, please contact John Shields at jshields-at-cb.uga.edu (706-542-4080), or to Bill Monroe at monroe-at-emcenter.msstate.edu (662-325- 3019). John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602
706-542-4080
==============================Original Headers============================== 4, 22 -- From jpshield-at-uga.edu Mon Jun 5 09:24:38 2006 4, 22 -- Received: from puntd5.cc.uga.edu (puntd5.cc.uga.edu [128.192.1.108]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k55EObbx001319 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 5 Jun 2006 09:24:38 -0500 4, 22 -- Received: from punts4.cc.uga.edu (punts4.cc.uga.edu [128.192.1.115]) 4, 22 -- by puntd5.cc.uga.edu (MOS 3.7.4b-GA) 4, 22 -- with ESMTP id CWW21100; 4, 22 -- Mon, 5 Jun 2006 10:24:36 -0400 (EDT) 4, 22 -- Received: (from punts4.cc.uga.edu [128.192.63.198]) 4, 22 -- by punts4.cc.uga.edu (MOS 3.7.4b-GA) 4, 22 -- with HTTPS/1.1 id AVU01645 (AUTH jpshield-at-uga.edu); 4, 22 -- Mon, 5 Jun 2006 10:24:34 -0400 (EDT) 4, 22 -- From: John Shields {jpshield-at-uga.edu} 4, 22 -- Subject: volunteer workers for M&M Chicago 4, 22 -- To: msa listserve {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Cc: {monroe-at-emcenter.msstate.edu} 4, 22 -- X-Mailer: Mirapoint Webmail Direct 3.7.4b-GA 4, 22 -- MIME-Version: 1.0 4, 22 -- Content-Type: text/plain; charset=us-ascii 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- Message-Id: {20060605102434.AVU01645-at-punts4.cc.uga.edu} 4, 22 -- Date: Mon, 5 Jun 2006 10:24:34 -0400 (EDT) ==============================End of - Headers==============================
We have a grad student user who is interested in thin sectioning carbon nanotubes. The tubes are in clusters and suspended in some buffer. Is it possible to embed the clumps in epoxy resin and section them? Is diamond knife going to survive after sectioning nanotubes? The average diameter of those tubes is approx. 1.4 nm. Any suggestion will be greatly appreciated.
Thanks in advance,
Soumitra
****************************************************************************** Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282 http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu
==============================Original Headers============================== 5, 20 -- From sghoshro-at-nmsu.edu Mon Jun 5 14:13:53 2006 5, 20 -- Received: from cheech.nmsu.edu (cheech.nmsu.edu [128.123.34.14]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k55JDrU9019226 5, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 5 Jun 2006 14:13:53 -0500 5, 20 -- Received: (from nobody-at-localhost) 5, 20 -- by cheech.nmsu.edu (8.11.6/8.11.6) id k55JDrU09264 5, 20 -- for Microscopy-at-microscopy.com; Mon, 5 Jun 2006 13:13:53 -0600 5, 20 -- Received: from 128.123.105.122 ( [128.123.105.122]) 5, 20 -- as user sghoshro-at-imap.nmsu.edu by webmail.nmsu.edu with HTTP; 5, 20 -- Mon, 5 Jun 2006 13:13:53 -0600 5, 20 -- Message-ID: {1149534833.44848271153f4-at-webmail.nmsu.edu} 5, 20 -- Date: Mon, 5 Jun 2006 13:13:53 -0600 5, 20 -- From: Soumitra Ghoshroy {sghoshro-at-nmsu.edu} 5, 20 -- To: Microscopy-at-microscopy.com 5, 20 -- Subject: sectioning carbon nanotubes 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; charset=ISO-8859-1 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- User-Agent: Internet Messaging Program (IMP) 3.0 5, 20 -- X-Originating-IP: 128.123.105.122 ==============================End of - Headers==============================
We are planning to examine some cellulose made microspheres containing albumin with transmission EM. Is there anybody experienced with this material before? Are routine EM tissue processing methods suitable with this material? Thanks for any kind of helps...
Dr. Necat Yilmaz Mersin Universitesi Tip Fakultesi Histoloji ve Embriyoloji Anabilim Dali
==============================Original Headers============================== 4, 31 -- From nyilmaz-at-mersin.edu.tr Tue Jun 6 07:32:34 2006 4, 31 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 4, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k56CWVBm020888 4, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 07:32:34 -0500 4, 31 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 4, 31 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 2AFA245146 4, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 15:32:34 +0300 (EEST) 4, 31 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 4, 31 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 4, 31 -- with ESMTP id 86250-05 for {Microscopy-at-microscopy.com} ; 4, 31 -- Tue, 6 Jun 2006 15:32:21 +0300 (EEST) 4, 31 -- Received: from NEJAT1 (unknown [193.255.128.130]) 4, 31 -- by mail.mersin.edu.tr (Postfix) with SMTP id 176D84513A 4, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 15:32:17 +0300 (EEST) 4, 31 -- Message-ID: {002e01c68965$3c9f8fb0$5201a8c0-at-NEJAT1} 4, 31 -- From: "Nejat" {nyilmaz-at-mersin.edu.tr} 4, 31 -- To: "EM-Mail Group" {Microscopy-at-microscopy.com} 4, 31 -- Subject: Help for cellulose microsphere processing 4, 31 -- Date: Tue, 6 Jun 2006 15:32:06 +0300 4, 31 -- MIME-Version: 1.0 4, 31 -- Content-Type: text/plain; 4, 31 -- format=flowed; 4, 31 -- charset="windows-1254"; 4, 31 -- reply-type=original 4, 31 -- Content-Transfer-Encoding: 7bit 4, 31 -- X-Priority: 3 4, 31 -- X-MSMail-Priority: Normal 4, 31 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 4, 31 -- Disposition-Notification-To: "Nejat" {nyilmaz-at-mersin.edu.tr} 4, 31 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 4, 31 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr ==============================End of - Headers==============================
We solved our contrasting problems by just following simple rules found in this list. If it solved our problems, I thought it could be useful for others.
- We prepared "carbonated lead citrate" (the time required to "burn" it was much shorter as proposed though) - We bought decarbonated NaOH to a big american supplier
These are very simple rules which make all the difference between "I see nothing" and "oh my god I never had such a nice staining". And that's true: I've never ever had such a beautiful staining!
Thank you again for your support.
Stéphane
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==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Tue Jun 6 09:10:06 2006 7, 18 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.87.65]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k56EA6M4002486 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 09:10:06 -0500 7, 18 -- Received: (qmail 2845 invoked by uid 60001); 6 Jun 2006 14:10:06 -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=nySdMzGji0kCEsaiYe3tpvougj/SVOO9ewwIWzMh2PBKN2eoYk1WF8iHp+JGNMcn+ly4RP7tBbHr5wlMS6iU4P42BsOTjHxoB1QGGLAU7XQL7doeX+fDLkwpvBBIcPd7VR0Rka5PTOGHmO3EMGBktWvXGdeK5WnyfMEqD6FpV5o= ; 7, 18 -- Message-ID: {20060606141006.2837.qmail-at-web37412.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Tue, 06 Jun 2006 07:10:06 PDT 7, 18 -- Date: Tue, 6 Jun 2006 07:10:06 -0700 (PDT) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: Lead citrate staining: Thank you again and again.... 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==============================
Here is another weird idea from me :-D I would like to localize a fluorescent molecule in the transversal plane of a cell monolayer(not from above). We have no material for the preparation of histological sections, but we have all necessary for TEM. I wonder if I could not embed the monolayer in Epon, then cut transversal semi-thin sections and observe in epifluroscence. Would Epon hinder fluorescence? Could it be done in another resin than Epon?
Regards,
Stéphane
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 6, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5783Hql015897 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 -0500 6, 18 -- Received: (qmail 21951 invoked by uid 60001); 7 Jun 2006 08:03:17 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 18 -- b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2Mk7fPbVFk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6XsGrL3IO0Qs+45yxbbeZT0= ; 6, 18 -- Message-ID: {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 01:03:17 PDT 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: weird idea: fluorescence in Epon 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==============================
I would appreciate any comments, references, or first-hand experiences from those using a combined FIB/SEM+Cryo instrument with & without a field emission gun - where biological or polymer samples can be sliced by the ion beam then imaged. I would be interested to know things such as:
Why you chose the instrument you use? How easy is it in practice to do this sort of ion beam slicing & is it restricted to particular types of biological or polymer samples? Does the FIB mode impact on the column cleanliness & resolution over time particularly if used to thin samples for TEM? Is the instrument expensive to run & does it require an experienced operator? Anything else you feel would help those looking at this type of instrument for a multi-user facility.
Many thanks Ursula -------------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
==============================Original Headers============================== 6, 23 -- From U.J.Potter-at-bath.ac.uk Wed Jun 7 05:11:33 2006 6, 23 -- Received: from binda.bath.ac.uk (binda.bath.ac.uk [138.38.32.22]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57ABWSl029055 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 05:11:33 -0500 6, 23 -- Received: from mary.bath.ac.uk ([138.38.32.14] ident=mmdf) 6, 23 -- by binda.bath.ac.uk with smtp id 1Fnv0e-0001sg-A3 6, 23 -- for microscopy-at-microscopy.com 6, 23 -- (return-path {U.J.Potter-at-bath.ac.uk} ); Wed, 07 Jun 2006 11:11:32 +0100 6, 23 -- Received: from eapc-03.campus.bath.ac.uk 6, 23 -- ( eapc-03.campus.bath.ac.uk [138.38.136.63] ) by bath.ac.uk 6, 23 -- id ab27759 for {microscopy-at-microscopy.com} ; 7 Jun 2006 11:11 +0100 6, 23 -- Date: Wed, 07 Jun 2006 11:11:24 +0100 6, 23 -- From: Ursula Potter {U.J.Potter-at-bath.ac.uk} 6, 23 -- To: microscopy-at-microscopy.com 6, 23 -- Subject: FIB/SEM+Cryo 6, 23 -- Message-ID: {7688765.1149678684-at-eapc-03.campus.bath.ac.uk} 6, 23 -- Originator-Info: login-id=mssujp; server=imaphost.bath.ac.uk 6, 23 -- X-Mailer: Mulberry/3.1.0 (Win32) 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; charset=us-ascii; FORMAT=flowed 6, 23 -- Content-Transfer-Encoding: 7bit 6, 23 -- Content-Disposition: inline 6, 23 -- X-Scanner: 207b733ce87962cd23b7af89110027a7b5b8d829 ==============================End of - Headers==============================
Are there anyone out there using this model? I would need specially the low level protocol on commanding it's RS232C port for lab automation purposes on our lab set-up at the university. Your help is greatly appreciated.
Regards,
Berns B.
-- -------------------------------------------- Bernardino Jerez Buenaobra University Research Associate National Institute of Physics University of the Philippines Diliman Campus 1101 Quezon City Philippines VOIP : +6329205301-99 local 3703 Fax/Data: +6329280296 URL: http://www.nip.upd.edu.ph/ipl email: bbuenaobra-at-nip.upd.edu.ph --------------------------------------------------------------------------
==============================Original Headers============================== 6, 31 -- From bbuenaobra-at-nip.upd.edu.ph Wed Jun 7 07:03:13 2006 6, 31 -- Received: from mail01.up.edu.ph (ns01.up.edu.ph [202.92.128.248]) 6, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k57C3Cmo009072 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 07:03:12 -0500 6, 31 -- Received: (qmail 26698 invoked by uid 510); 7 Jun 2006 12:04:37 -0000 6, 31 -- Received: from 10.16.3.34 by web1 (envelope-from {bbuenaobra-at-nip.upd.edu.ph} , uid 501) with qmail-scanner-1.25 6, 31 -- (clamav 15 Apr 2006 09-05 +0200 6, 31 -- Clear:RC:1(10.16.3.34):. 6, 31 -- Processed in 84.072928 secs); 07 Jun 2006 12:04:37 -0000 6, 31 -- Received: from unknown (HELO web4.up.edu.ph) (10.16.3.34) 6, 31 -- by 10.16.3.142 with SMTP; 7 Jun 2006 12:03:10 -0000 6, 31 -- Received: (qmail 1210 invoked by uid 1008); 7 Jun 2006 11:55:47 -0000 6, 31 -- Received: from 10.32.148.2 by web4 (envelope-from {bbuenaobra-at-nip.upd.edu.ph} , uid 1002) with qmail-scanner-1.25 6, 31 -- (clamav 12 Oct 2005 22-35 +0200 6, 31 -- Clear:RC:1(10.32.148.2):. 6, 31 -- Processed in 1.24726 secs); 07 Jun 2006 11:55:47 -0000 6, 31 -- Received: from unknown (HELO nip.upd.edu.ph) (root-at-10.32.148.2) 6, 31 -- by 10.16.3.34 with SMTP; 7 Jun 2006 11:55:46 -0000 6, 31 -- Received: from [192.168.101.101] (suez.nip.upd.edu.ph [192.168.115.13]) 6, 31 -- by nip.upd.edu.ph (8.9.3/8.9.3) with ESMTP id SAA17852 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 18:18:40 -0400 6, 31 -- Message-ID: {4487879B.2000600-at-nip.upd.edu.ph} 6, 31 -- Date: Wed, 07 Jun 2006 18:12:43 -0800 6, 31 -- From: "Bernardino J. Buenaobra" {bbuenaobra-at-nip.upd.edu.ph} 6, 31 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 6, 31 -- X-Accept-Language: en-us, en 6, 31 -- MIME-Version: 1.0 6, 31 -- To: Microscopy-at-microscopy.com 6, 31 -- Subject: Hamamatsu C5985 Chilled CCD Camera RS232 Interface 6, 31 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 31 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Stephane, Not so weird, but might not work. The two things to worry about are autofluorescence and loss of your fluorochrome. If you have some of your cells (unlabeled) in epon you can check some sections under a fluorescence microscope and see about the autofuor. I think the loss of fluorescence is a more serious problem, either because it is extracted during dehydration/infiltration or quenched by the embedment. You may have to embed a labeled sample and find out the hard way!
Tobias } } Dear listers, } } Here is another weird idea from me :-D } I would like to localize a fluorescent molecule in the } transversal plane of a cell monolayer(not from above). } We have no material for the preparation of } histological sections, but we have all necessary for } TEM. I wonder if I could not embed the monolayer in } Epon, then cut transversal semi-thin sections and } observe in epifluroscence. } Would Epon hinder fluorescence? } Could it be done in another resin than Epon? } } Regards, } } StČphane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 } 6, 18 -- Received: from } web37410.mail.mud.yahoo.com } (web37410.mail.mud.yahoo.com [209.191.87.63]) } 6, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k5783Hql015897 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 -0500 } 6, 18 -- Received: (qmail 21951 invoked by uid } 60001); 7 Jun 2006 08:03:17 -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 6, 18 -- } b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2Mk7fPbVFk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6XsGrL3IO0Qs+45yxbbeZT0= } ; } 6, 18 -- Message-ID: {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37410.mail.mud.yahoo.com via HTTP; Wed, 07 } Jun 2006 01:03:17 PDT } 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: weird idea: fluorescence in Epon } 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==============================
Regarding the idea of localizing fluorescence label in epon, I would suggest that you not use Epon...there are autoflourescence problems (at least in our hands, using Spurrs resin). However, I think you may be happy using LR White, polymerized at 60C. We've done some similar work here and have been very impressed with the results.
Good luck,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org -----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Wednesday, June 07, 2006 1:10 AM To: drk-at-SHCC.org
Dear listers,
Here is another weird idea from me :-D I would like to localize a fluorescent molecule in the transversal plane of a cell monolayer(not from above). We have no material for the preparation of histological sections, but we have all necessary for TEM. I wonder if I could not embed the monolayer in Epon, then cut transversal semi-thin sections and observe in epifluroscence. Would Epon hinder fluorescence? Could it be done in another resin than Epon?
Regards,
Stéphane
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 6, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5783Hql015897 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 -0500 6, 18 -- Received: (qmail 21951 invoked by uid 60001); 7 Jun 2006 08:03:17 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content -Transfer-Encoding; 6, 18 -- b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2Mk7fPbV Fk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6X sGrL3IO0Qs+45yxbbeZT0= ; 6, 18 -- Message-ID: {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 01:03:17 PDT 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: weird idea: fluorescence in Epon 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============================== 16, 22 -- From drk-at-SHCC.org Wed Jun 7 12:46:36 2006 16, 22 -- Received: from shcc.org (mail.shcc.org [64.213.211.200]) 16, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57HkZdU005537 16, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 7 Jun 2006 12:46:35 -0500 16, 22 -- Received: from DRK2 ("port 1395"-at-[64.213.211.3]) by SHCC.org (PMDF V6.1 #37411) 16, 22 -- with ESMTPA id {01M3CA5GN1460004DI-at-SHCC.org} for Microscopy-at-Microscopy.Com; 16, 22 -- Wed, 07 Jun 2006 10:50:12 -0800 (PST) 16, 22 -- Date: Wed, 07 Jun 2006 10:43:17 -0700 16, 22 -- From: Doug Keene {drk-at-SHCC.org} 16, 22 -- Subject: RE: [Microscopy] weird idea: fluorescence in Epon 16, 22 -- In-reply-to: {200606070809.k5789kOv024103-at-ns.microscopy.com} 16, 22 -- To: Microscopy-at-Microscopy.Com 16, 22 -- Reply-to: drk-at-SHCC.org 16, 22 -- Message-id: {01M3CA5GPO8O0004DI-at-SHCC.org} 16, 22 -- Organization: Shriners Hospitals for Children 16, 22 -- MIME-version: 1.0 16, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 16, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 16, 22 -- Content-type: text/plain; charset=iso-8859-1 16, 22 -- Thread-Index: AcaKCQiVuqpcDFhbQBCvgNSUNvkSUwAUE+Gg 16, 22 -- Content-Transfer-Encoding: 8bit 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k57HkZdU005537 ==============================End of - Headers==============================
Don't ever hesitate. some of my best things have happened because of an offbeat idea or insane shot in the dark question during discussions. Of course, I don't want to admit that to my wife. She hates to be referred to as an offbeat idea, just as the best thing that ever happened to me O:-) .
Frankly, you're keeping the rest of us fresh and on our toes.
Paul
==============================Original Headers============================== 4, 21 -- From paul_hazelton-at-umanitoba.ca Wed Jun 7 13:31:24 2006 4, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57IVNQJ016492 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 13:31:24 -0500 4, 21 -- Received: from [130.179.152.79] (cvx-016.cc.umanitoba.ca [130.179.152.79]) 4, 21 -- (authenticated bits=0) 4, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k57IVLmc025694; 4, 21 -- Wed, 7 Jun 2006 13:31:22 -0500 (CDT) 4, 21 -- Message-ID: {44871B76.7060704-at-umanitoba.ca} 4, 21 -- Date: Wed, 07 Jun 2006 13:31:18 -0500 4, 21 -- From: Paul Hazelton {paul_hazelton-at-umanitoba.ca} 4, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 4, 21 -- X-Accept-Language: en-us, en 4, 21 -- MIME-Version: 1.0 4, 21 -- To: nizets2-at-yahoo.com 4, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 4, 21 -- Subject: Re: [Microscopy] weird idea: fluorescence in Epon 4, 21 -- References: {200606070806.k5786Xow019680-at-ns.microscopy.com} 4, 21 -- In-Reply-To: {200606070806.k5786Xow019680-at-ns.microscopy.com} 4, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 21 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Dear without an I, Success will depend upon the fluorophore. FITC and TRITC will lose their fluorescence. Alexa fluor dyes and cyanine dyes will work well. Actually, I think with reduced photobleaching. Autofluorescence will depend upon imaging modality. Spurr's looks great under the confocal, but the autolfuorescence creates an impossible haze with epi-fluorescence on thick samples. Deconvolution will clean up the haze. 3 micron sections looked very good. the new formulation of spurr's is too brittle so I'm going to try Eponate 812 or something similar for my next round of embedded fluorescence. I tried Histo-Resin and found the autofluorescence was noticeable under confocal, but didn't try LR-White.
Regards, Glen On Jun 7, 2006, at 10:49 AM, drk-at-SHCC.org wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Regarding the idea of localizing fluorescence label in epon, I } would suggest } that you not use Epon...there are autoflourescence problems (at } least in our } hands, using Spurrs resin). However, I think you may be happy } using LR } White, polymerized at 60C. We've done some similar work here and } have been } very impressed with the results. } } Good luck, } } Doug } } Douglas R. Keene } Assistant Investigator } Micro-Imaging Center } Shriners Hospital for Children } 3102 S.W. Sam Jackson Park Road } Portland, Oregon 97239 } 503-221-3434 } drk-at-shcc.org } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Wednesday, June 07, 2006 1:10 AM } To: drk-at-SHCC.org } Subject: [Microscopy] weird idea: fluorescence in Epon } } } } } ---------------------------------------------------------------------- } ------ } 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, } } Here is another weird idea from me :-D } I would like to localize a fluorescent molecule in the } transversal plane of a cell monolayer(not from above). } We have no material for the preparation of } histological sections, but we have all necessary for } TEM. I wonder if I could not embed the monolayer in } Epon, then cut transversal semi-thin sections and } observe in epifluroscence. } Would Epon hinder fluorescence? } Could it be done in another resin than Epon? } } Regards, } } Stéphane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 } 6, 18 -- Received: from web37410.mail.mud.yahoo.com } (web37410.mail.mud.yahoo.com [209.191.87.63]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k5783Hql015897 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 } -0500 } 6, 18 -- Received: (qmail 21951 invoked by uid 60001); 7 Jun 2006 } 08:03:17 } -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content- } Type:Content } -Transfer-Encoding; } 6, 18 -- } b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2M } k7fPbV } Fk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/ } 0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6X } sGrL3IO0Qs+45yxbbeZT0= ; } 6, 18 -- Message-ID: } {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37410.mail.mud.yahoo.com via } HTTP; Wed, 07 Jun 2006 01:03:17 PDT } 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: weird idea: fluorescence in Epon } 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============================== } 16, 22 -- From drk-at-SHCC.org Wed Jun 7 12:46:36 2006 } 16, 22 -- Received: from shcc.org (mail.shcc.org [64.213.211.200]) } 16, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k57HkZdU005537 } 16, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 7 Jun 2006 } 12:46:35 -0500 } 16, 22 -- Received: from DRK2 ("port 1395"-at-[64.213.211.3]) by } SHCC.org (PMDF V6.1 #37411) } 16, 22 -- with ESMTPA id {01M3CA5GN1460004DI-at-SHCC.org} for } Microscopy-at-Microscopy.Com; } 16, 22 -- Wed, 07 Jun 2006 10:50:12 -0800 (PST) } 16, 22 -- Date: Wed, 07 Jun 2006 10:43:17 -0700 } 16, 22 -- From: Doug Keene {drk-at-SHCC.org} } 16, 22 -- Subject: RE: [Microscopy] weird idea: fluorescence in Epon } 16, 22 -- In-reply-to: {200606070809.k5789kOv024103-at-ns.microscopy.com} } 16, 22 -- To: Microscopy-at-Microscopy.Com } 16, 22 -- Reply-to: drk-at-SHCC.org } 16, 22 -- Message-id: {01M3CA5GPO8O0004DI-at-SHCC.org} } 16, 22 -- Organization: Shriners Hospitals for Children } 16, 22 -- MIME-version: 1.0 } 16, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 16, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 } 16, 22 -- Content-type: text/plain; charset=iso-8859-1 } 16, 22 -- Thread-Index: AcaKCQiVuqpcDFhbQBCvgNSUNvkSUwAUE+Gg } 16, 22 -- Content-Transfer-Encoding: 8bit } 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k57HkZdU005537 } ==============================End of - } Headers==============================
==============================Original Headers============================== 6, 25 -- From glenmac-at-u.washington.edu Wed Jun 7 15:36:08 2006 6, 25 -- Received: from mxout7.cac.washington.edu (mxout7.cac.washington.edu [140.142.32.178]) 6, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57Ka8xv029950 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 15:36:08 -0500 6, 25 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 6, 25 -- by mxout7.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k57Ka6Qu010939 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 13:36:07 -0700 6, 25 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 6, 25 -- (authenticated authid=glenmac) 6, 25 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k57Ka6FG007766 6, 25 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 13:36:06 -0700 6, 25 -- Mime-Version: 1.0 (Apple Message framework v750) 6, 25 -- In-Reply-To: {200606071749.k57HnXYf008396-at-ns.microscopy.com} 6, 25 -- References: {200606071749.k57HnXYf008396-at-ns.microscopy.com} 6, 25 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 6, 25 -- Message-Id: {B6197299-29B8-44FB-9547-C7E54AE7E765-at-u.washington.edu} 6, 25 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 6, 25 -- Subject: RE: weird idea 6, 25 -- Date: Wed, 7 Jun 2006 13:36:03 -0700 6, 25 -- To: microscopy-at-microscopy.com 6, 25 -- X-Mailer: Apple Mail (2.750) 6, 25 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='IP_HTTP_ADDR 0, __C230066_P5 0, __CP_NAME_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_FUNWORDS 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0, __STOCK_PHRASE_24 0' 6, 25 -- Content-Transfer-Encoding: 8bit 6, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k57Ka8xv029950 ==============================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 wim.vandenbroeck-at-UGent.be as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: wim.vandenbroeck-at-UGent.be Name: Wim Van den Broeck
Organization: Morphology, Ghent University
Title-Subject: [Filtered] TEM - virus particles in cell culture
Question: Dear Friends,
I need to visualize viral particles grown in cell culture, but I do not have any experience with that (only with tissue samples). Could you give me any advice (collection of cells, fixation, embedding, Ö.).
Thanks in advance,
Wim Van den Broeck.
Wim Van den Broeck, DVM, MSc, PhD Professor in Cytology and Histology Department of Morphology Faculty of Veterinary Medicine, Ghent University Salisburylaan 133, B-9820 Merelbeke, BELGIUM tel.: +32 (0)9 264 77 16 fax: +32 (0)9 264 77 90 Email: wim.vandenbroeck-at-UGent.be
While on the Medical School Faculty of USC, Los Angeles I did extensive studies of hepatitis B virus in liver explants. The technique is straightforward.
Remove cells using a soft spatula or a pipette depending on whether or not the cells are attached to the surface of the culture container.
Place the cells in a centrifuge tube containg fixative. Carry out the dehydration and embedding fluid impregnation process in the centrifuge tube spinning between each stage. Finally transfer the embedding mixture with the cells into a Beem-type embedding capsule and spin the cells down into the tip.
If you wish I can send you the reference for one of my publications that outlines the techniques employed.
Best wishes,
Ted Dunn The EMscope Company Ltd. Thailand
--- wim.vandenbroeck-at-UGent.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 } ---------------------------------------------------------------------------- } } 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 wim.vandenbroeck-at-UGent.be as well } as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: wim.vandenbroeck-at-UGent.be } Name: Wim Van den Broeck } } Organization: Morphology, Ghent University } } Title-Subject: [Filtered] TEM - virus particles in } cell culture } } Question: Dear Friends, } } I need to visualize viral particles grown in cell } culture, but I do not have any experience with that } (only with tissue samples). Could you give me any } advice (collection of cells, fixation, embedding, } Ö.). } } Thanks in advance, } } Wim Van den Broeck. } } Wim Van den Broeck, DVM, MSc, PhD } Professor in Cytology and Histology } Department of Morphology } Faculty of Veterinary Medicine, Ghent University } Salisburylaan 133, B-9820 Merelbeke, } BELGIUM } tel.: +32 (0)9 264 77 16 } fax: +32 (0)9 264 77 90 } Email: wim.vandenbroeck-at-UGent.be } } } } --------------------------------------------------------------------------- } } } ==============================Original } Headers============================== } 13, 14 -- From zaluzec-at-microscopy.com Wed Jun 7 } 19:36:50 2006 } 13, 14 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 13, 14 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k580anBX013138 } 13, 14 -- for {microscopy-at-microscopy.com} ; Wed, 7 } Jun 2006 19:36:50 -0500 } 13, 14 -- Mime-Version: 1.0 } 13, 14 -- X-Sender: (Unverified) } 13, 14 -- Message-Id: } {p06110401c0ad218d9302-at-[206.69.208.22]} } 13, 14 -- Date: Wed, 7 Jun 2006 19:36:48 -0500 } 13, 14 -- To: microscopy-at-microscopy.com } 13, 14 -- From: wim.vandenbroeck-at-UGent.be (by way of } MicroscopyListserver) } 13, 14 -- Subject: viaWWW: TEM - virus particles in } cell culture } 13, 14 -- Content-Type: text/plain; } charset="iso-8859-1" } 13, 14 -- Content-Transfer-Encoding: 8bit } 13, 14 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } k580anBX013138 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 12, 20 -- From drteddunne-at-yahoo.com Wed Jun 7 23:57:20 2006 12, 20 -- Received: from web33409.mail.mud.yahoo.com (web33409.mail.mud.yahoo.com [68.142.206.141]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k584vKhx028460 12, 20 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 23:57:20 -0500 12, 20 -- Received: (qmail 54631 invoked by uid 60001); 8 Jun 2006 04:57:19 -0000 12, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 12, 20 -- s=s1024; d=yahoo.com; 12, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 12, 20 -- b=ii5iIj/zzCWEX2M/ngtTuKWaGi8Z+F/IefIeH2PH3lxLPeW1epwoduWCMBPKHXXdMNBhf+QT33GPjKZYFC7TkD2ZIvkyySuyJZb+Ee3LkP4ReWtMu81pX1SxE14EKdXGKwtDlOvan5i90MACA9VNLQLULr+BoQvcXeaygnrVzIk= ; 12, 20 -- Message-ID: {20060608045719.54629.qmail-at-web33409.mail.mud.yahoo.com} 12, 20 -- Received: from [202.47.247.136] by web33409.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 21:57:19 PDT 12, 20 -- Date: Wed, 7 Jun 2006 21:57:19 -0700 (PDT) 12, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: TEM - virus particles in cell culture 12, 20 -- To: wim.vandenbroeck-at-UGent.be 12, 20 -- Cc: microscopy-at-microscopy.com 12, 20 -- In-Reply-To: {200606080040.k580eZFL018920-at-ns.microscopy.com} 12, 20 -- MIME-Version: 1.0 12, 20 -- Content-Type: text/plain; charset=iso-8859-1 12, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
It depends if you need to visualize the viral particles inside the cells or not. If so, I would consider usual protocol for classical cell morphology.
I already observed viral particles in cells I observed for other purposes. If you need the viral particles alone, you have 2 solutions depending on the concentration of the virus in medium.
1) If it is concentrated, you just collect the supernatant, centrifuge 5 min at 5000 RPM to pellet the cells and cell debris, and collect the supernatant again.
2) If it is diluted, you have to find an ultrafast centrifuge and perform an additional centrifugation to pellet your virus particles and concentrate them.
Then you can just do a negative staining (PTA worked well with rhinoviruses for me). You will find tons of protocols on the net.
Good luck.
Stephane
--- wim.vandenbroeck-at-UGent.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 } ---------------------------------------------------------------------------- } } 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 wim.vandenbroeck-at-UGent.be as well } as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: wim.vandenbroeck-at-UGent.be } Name: Wim Van den Broeck } } Organization: Morphology, Ghent University } } Title-Subject: [Filtered] TEM - virus particles in } cell culture } } Question: Dear Friends, } } I need to visualize viral particles grown in cell } culture, but I do not have any experience with that } (only with tissue samples). Could you give me any } advice (collection of cells, fixation, embedding, } Ö.). } } Thanks in advance, } } Wim Van den Broeck. } } Wim Van den Broeck, DVM, MSc, PhD } Professor in Cytology and Histology } Department of Morphology } Faculty of Veterinary Medicine, Ghent University } Salisburylaan 133, B-9820 Merelbeke, } BELGIUM } tel.: +32 (0)9 264 77 16 } fax: +32 (0)9 264 77 90 } Email: wim.vandenbroeck-at-UGent.be } } } } --------------------------------------------------------------------------- } } } ==============================Original } Headers============================== } 13, 14 -- From zaluzec-at-microscopy.com Wed Jun 7 } 19:36:50 2006 } 13, 14 -- Received: from [206.69.208.22] } (mac22.zaluzec.com [206.69.208.22]) } 13, 14 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k580anBX013138 } 13, 14 -- for {microscopy-at-microscopy.com} ; Wed, 7 } Jun 2006 19:36:50 -0500 } 13, 14 -- Mime-Version: 1.0 } 13, 14 -- X-Sender: (Unverified) } 13, 14 -- Message-Id: } {p06110401c0ad218d9302-at-[206.69.208.22]} } 13, 14 -- Date: Wed, 7 Jun 2006 19:36:48 -0500 } 13, 14 -- To: microscopy-at-microscopy.com } 13, 14 -- From: wim.vandenbroeck-at-UGent.be (by way of } MicroscopyListserver) } 13, 14 -- Subject: viaWWW: TEM - virus particles in } cell culture } 13, 14 -- Content-Type: text/plain; } charset="iso-8859-1" } 13, 14 -- Content-Transfer-Encoding: 8bit } 13, 14 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } k580anBX013138 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 13, 20 -- From nizets2-at-yahoo.com Thu Jun 8 01:33:06 2006 13, 20 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k586X6Rv008226 13, 20 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 01:33:06 -0500 13, 20 -- Received: (qmail 55709 invoked by uid 60001); 8 Jun 2006 06:33:05 -0000 13, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 13, 20 -- s=s1024; d=yahoo.com; 13, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 13, 20 -- b=Fsb/2635sW1xpqYSJnpNsxRuXjWSQC4ZSMEQFn3Yquyv/aPwzxX0FRbUXkLKZbkhxW68SgzbmFvBvPPlenBsbYoFrfcTMqb1AEv3c+TGRtOO04pJOKy2iFrdunIgjwMtvkzWuNbp9OESpfyRU5/X8aoheanFV5OzQL4k8yq9fWU= ; 13, 20 -- Message-ID: {20060608063305.55707.qmail-at-web37410.mail.mud.yahoo.com} 13, 20 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 23:33:05 PDT 13, 20 -- Date: Wed, 7 Jun 2006 23:33:05 -0700 (PDT) 13, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 13, 20 -- Subject: Re: [Microscopy] viaWWW: TEM - virus particles in cell culture 13, 20 -- To: wim.vandenbroeck-at-UGent.be 13, 20 -- Cc: microscopy-at-microscopy.com 13, 20 -- In-Reply-To: {200606080042.k580gPDu021981-at-ns.microscopy.com} 13, 20 -- MIME-Version: 1.0 13, 20 -- Content-Type: text/plain; charset=iso-8859-1 13, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
For those interested in the technique, it is simply available in the internet site of EMS. Look in Technical-} tips and articles--} "A stable lead staining solution".
For the very practical side of the preparation of carbonated lead citrate, here is what I exactly did: I heated 2g at 250°C for 3h in a ceramic beaker (called "melting pot" on EMS site) with a glass cover. I have the chance to work with chemists so the equipment is not a problem: I used a temperature-controlled special oven connected to a mobile hood.
I controlled the color of the powder. It turns grey, then yellowish grey, a little bit like humid sand. I stopped then. If one is unsure, one can still heat several beakers each containing 1g of lead citrate and heat them for different times (2h, 3h, 4h).
I think it is pretty straithforward because it worked the first time I tried. I stored the powder in a sealed glass flask (don't know if it is the best though).
Now IMHO, seeing the easiness of the technique and the cheap price of carbonate-free NaOH, there is really no reason NOT to do it :-D
Stephane
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==============================Original Headers============================== 8, 19 -- From nizets2-at-yahoo.com Thu Jun 8 01:55:43 2006 8, 19 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.87.65]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k586thms018635 8, 19 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 01:55:43 -0500 8, 19 -- Received: (qmail 55159 invoked by uid 60001); 8 Jun 2006 06:55:43 -0000 8, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 19 -- s=s1024; d=yahoo.com; 8, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 19 -- b=HSwCug+LY+IF9G7UfVpv3nug1PPUOuKu932Mci/MLXwwynTrFZBvG/o4Fi3NLhxTH7BOceQ0qBQfwoAbazPN3jdO08n6Ux3BTmfGzWngftdTk9q0wgb4vzW4h+aWlkz5SlhxyofQMtJimVunPbuxbr/q6gUn8x9QmTJmDfyIHW8= ; 8, 19 -- Message-ID: {20060608065543.55157.qmail-at-web37412.mail.mud.yahoo.com} 8, 19 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 23:55:43 PDT 8, 19 -- Date: Wed, 7 Jun 2006 23:55:43 -0700 (PDT) 8, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 19 -- Subject: Re: [Microscopy] Lead citrate staining: Thank you again and again.... 8, 19 -- To: microscopy-at-microscopy.com 8, 19 -- In-Reply-To: {p0621021bc0acd7188676-at-[10.0.1.3]} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: text/plain; charset=iso-8859-1 8, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Thank you for your numerous answers. Lots of them show a concern about autofluorescence, but I must say that I was more concerned about the quenching by Epon and the processing steps: dehydration and embedding. I wondered if and how it could "damage" the fluorochrome.
We use Epon 812 (glycidether 100) and we just cut empty block at 300 to 500nm thickness to observe the autofluorescence with the "green" filter (our fluorochrome is Alexa488). Actually there IS some autofluorescence, but not dramatic even with a thickness of 500nm. I don't think it will perturb the observation, but it depends of course how well the alexa will sustain the processing.
An excellent remark was made, pointing out that glutaraldehyde must be avoided because it brings autofluorescence. I knew it but recalling me was not bad :-D Anyway glutar is not necessary for LM observation.
I will definitely try light fixation (4% PFA for 20 min), fast dehydration in alcohol and direct embedding in Epon.
Another concern is the localisation of the cells and cell comparments. Do some of you have an idea how well work DIC or phase contrast with cells embedded in Epon? Could I just use general staining protocols hematoxylin-eosin before embedding in Epon?
Stephane
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==============================Original Headers============================== 11, 19 -- From nizets2-at-yahoo.com Thu Jun 8 07:47:23 2006 11, 19 -- Received: from web37402.mail.mud.yahoo.com (web37402.mail.mud.yahoo.com [209.191.87.55]) 11, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58ClNTT023147 11, 19 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 07:47:23 -0500 11, 19 -- Received: (qmail 73767 invoked by uid 60001); 8 Jun 2006 12:47:22 -0000 11, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 19 -- s=s1024; d=yahoo.com; 11, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 11, 19 -- b=iV+QTQg28QV+UWQjdDyBjy4AJOhD8LlQroQUt++DI1fpwtngYP1g0BTye9C09Ku/OrUlcNGCcYXn305nvBYPPrCCh5x8htA4EZTmi/om8WXtIK2/iQjH179gPv46rnhLUg/D+9WRqfRTh7T0s4plthEYocx1ZegPqnfIsczbFfE= ; 11, 19 -- Message-ID: {20060608124722.73765.qmail-at-web37402.mail.mud.yahoo.com} 11, 19 -- Received: from [80.122.101.102] by web37402.mail.mud.yahoo.com via HTTP; Thu, 08 Jun 2006 05:47:22 PDT 11, 19 -- Date: Thu, 8 Jun 2006 05:47:22 -0700 (PDT) 11, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 11, 19 -- Subject: Re: [Microscopy] RE: weird idea 11, 19 -- To: microscopy-at-microscopy.com 11, 19 -- In-Reply-To: {200606072041.k57KfLwR006361-at-ns.microscopy.com} 11, 19 -- MIME-Version: 1.0 11, 19 -- Content-Type: text/plain; charset=iso-8859-1 11, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Dear Stephane, Don't use eosin...it fluoresces very intensely (in the usual "Rhodamine" wavelengths). In fact, the slide I use when teaching people how to use the confocal is a standard paraffin section stained with H&E. It never seems to bleach, and it fluoresces like mad at 488 (the connective tissue), 543, and 633.
I've looked at semi-thin resin sections with DIC...it works. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 2, 23 -- From lcgould-at-med.cornell.edu Thu Jun 8 08:28:44 2006 2, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 2, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58DSgdm001635 2, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 08:28:43 -0500 2, 23 -- Received: from mpx2.med.cornell.edu (pc113142-10.med.cornell.edu [140.251.11.119]) 2, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k58DSdim006316 2, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 09:28:41 -0400 (EDT) 2, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 2, 23 -- by mpx2.med.cornell.edu 2, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 2, 23 -- with ESMTPA id {0J0J00KT0MRQ1450-at-mpx2.med.cornell.edu} for 2, 23 -- microscopy-at-microscopy.com; Thu, 08 Jun 2006 09:28:39 -0400 (EDT) 2, 23 -- Date: Thu, 08 Jun 2006 09:21:47 -0400 2, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 2, 23 -- Subject: Re: [Microscopy] weird idea 2, 23 -- In-reply-to: {200606081248.k58Cm7jk024433-at-ns.microscopy.com} 2, 23 -- Sender: lcgould-at-med.cornell.edu 2, 23 -- To: nizets2-at-yahoo.com, Microscopy Listserver {microscopy-at-microscopy.com} 2, 23 -- Message-id: {p06230903c0add44359ef-at-[140.251.48.23]} 2, 23 -- MIME-version: 1.0 2, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 2, 23 -- References: {200606081248.k58Cm7jk024433-at-ns.microscopy.com} 2, 23 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.6.8.60432 ==============================End of - Headers==============================
I put my TEM negatives on a lightbox and photograph them with a Canon G3 digital camera. Then I convert to a positive with PhotoShop.
Geoff
aarti_harle-at-yahoo.co.in wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 9, 35 -- From mcauliff-at-umdnj.edu Thu Jun 8 08:35:24 2006 9, 35 -- Received: from zix04.umdnj.edu (zix04.UMDNJ.EDU [130.219.34.127]) 9, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58DZOFj011595 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:24 -0500 9, 35 -- Received: from zix04.umdnj.edu (ZixVPM [127.0.0.1]) 9, 35 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id C348C4BE51 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:23 -0500 (CDT) 9, 35 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 9, 35 -- by zix04.umdnj.edu (Proprietary) with ESMTP id 05B9F4BE6A 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:21 -0500 (CDT) 9, 35 -- Received: from ([130.219.34.131]) 9, 35 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.16981519; 9, 35 -- Thu, 08 Jun 2006 09:35:01 -0400 9, 35 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 9, 35 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 9, 35 -- id {0J0J00501N192Q-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 9, 35 -- for microscopy-at-msa.microscopy.com; Thu, 08 Jun 2006 09:35:01 -0400 (EDT) 9, 35 -- Received: from [127.0.0.1] ([10.138.2.123]) 9, 35 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 9, 35 -- 2004)) with ESMTP id {0J0J005AJN211E-at-Polaris.umdnj.edu} ; Thu, 9, 35 -- 08 Jun 2006 09:34:50 -0400 (EDT) 9, 35 -- Date: Thu, 08 Jun 2006 09:35:48 -0400 9, 35 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 9, 35 -- Subject: Re: [Microscopy] Scanner 9, 35 -- In-reply-to: {200606081144.k58BiOek013698-at-ns.microscopy.com} 9, 35 -- To: aarti_harle-at-yahoo.co.in, 9, 35 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 9, 35 -- Message-id: {448827B4.5000709-at-umdnj.edu} 9, 35 -- MIME-version: 1.0 9, 35 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 9, 35 -- Content-transfer-encoding: 7BIT 9, 35 -- X-Accept-Language: en-us, en 9, 35 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 9, 35 -- Gecko/20040804 Netscape/7.2 (ax) 9, 35 -- References: {200606081144.k58BiOek013698-at-ns.microscopy.com} ==============================End of - Headers==============================
I'll send a pdf of a short article on scanners I wrote for Microscopy Today (May 2006). My personal favourite scanner is the Epson V750 Pro (Ł600) and the Cheaper sister the Epson V700 (Ł400). The older model Epson 4990 Photo is not quite as good but cheaper at Ł280. All these scan film up to full A4 size, and have an easy to use twain interface. They also do reflective scans for paper and photographs. My favourite scanner review site for pro-sumer (i.e. cheap) scanners is http://www.photo-i.co.uk.
As mentioned you can use a camera and stand to copy film (used to be called an epidiascope in my Quantimet image analyser days) - and you can get pretty good optical microscope images just by resting a compact digital camera lens against the eyepiece. These days though I think for film, 6,400 dpi pro-sumer flatbed scanners do it better, more conveniently and relatively cheaply.
For image analysis there's lot to choose from, e.g. Improvision OpenLabs and Velocity (http://www.improvision.co.uk) ImageProPlus (http://www.mediacy.com) Kinetic Imaging (now http://www.andor.com/products) MetaMorph (http://www.moleculardevices.com/pages/software/metamorph.html)
These packages are expensive though (typically Ł3k+). They all deal with scanned tif images (and much more).
However if you don't have Ł3,000 for the above software, have at look at the freeware ImageJ (for the PC) at http://rsb.info.nih.gov/ij - you often need some of the plug-ins as well (there is also the sister program NIHImage for Apple fans at http://rsb.info.nih.gov/nih-image). Not always an easy program to get to grips with quickly (like most image analysis software), but quite powerful with plug-ins. For simple image manipulation, Photoshop CS2 is the obvious choice - otherwise the cut-down Photoshop Elements is pretty good and is supplied free with the above scanners (as a slightly older version).
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
I wonder if the autofluorescence might be due primarily to the MNA (aka NMA) in the usual Luft formulation?
When i found that NMA caused the cured resin to react horribly with MnO4 section stain --- (useful because the Mn04-Pb staining sequence gives much more contrast than any other stain we've ever tried) --- I learned to avoid MNA and just empirically readjusted DDSA and Epon ratios empirically for a firm enough cured resin, needing no MNA, yet compatible with MnO4 staining. However, I soon abandoned Epon and Spurr of all kinds in favor of an Araldite 506-DDSA-DER 736 mixture (10:15:2 gm by weight) because it sections thinner and accepts Mn-Pb staining with less graininess. So that Araldite mix is what i will try soon with Alexa 488-phalloidin.
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-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
Hi Mike, Interesting thought about the NMA, and your Araldite recipe looks worth a try. I had dropped the DMAE in Spurr's to 1/2 and then to 1/4 of the original recipe, thinking that it might generate radicals reacting with the fluorophores. The result was slightly reduced autofluorescence after a 3 day cure time at 60 C. No apparent effect on the fluorophores themselves, but didn't spend any time measuring.
FYI, alcohols will cause phalloidin to dissociate from the actin. Maybe it will survive a higher alcohol, propyl or butyl, but an actin antibody would be a better choice if needing to dehydrate the sample.
Regards, Glen
Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
On Jun 8, 2006, at 9:17 AM, Mike Reedy wrote:
} Wonderful weird idea! } } I wonder if the autofluorescence might be due primarily to the MNA } (aka NMA) in the usual Luft formulation? } } When i found that NMA caused the cured resin to react horribly with } MnO4 section stain --- (useful because the Mn04-Pb staining } sequence gives much more contrast than any other stain we've ever } tried) --- I learned to avoid MNA and just empirically readjusted } DDSA and Epon ratios empirically for a firm enough cured resin, } needing no MNA, yet compatible with MnO4 staining. However, I soon } abandoned Epon and Spurr of all kinds in favor of an Araldite 506- } DDSA-DER 736 mixture (10:15:2 gm by weight) because it sections } thinner and accepts Mn-Pb staining with less graininess. So that } Araldite mix is what i will try soon with Alexa 488-phalloidin. } } -mike reedy- } } --------------------------------------------------------------------- } } ------- } } 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 without an I, } } Success will depend upon the fluorophore. FITC and TRITC will lose } } their fluorescence. Alexa fluor dyes and cyanine dyes will work } } well. Actually, I think with reduced photobleaching. } } Autofluorescence will depend upon imaging modality. Spurr's looks } } great under the confocal, but the autolfuorescence creates an } } impossible haze with epi-fluorescence on thick samples. } } Deconvolution will clean up the haze. 3 micron sections looked very } } good. the new formulation of spurr's is too brittle so I'm going to } } try Eponate 812 or something similar for my next round of embedded } } fluorescence. I tried Histo-Resin and found the autofluorescence was } } noticeable under confocal, but didn't try LR-White. } } } } Hardie, MacDonald, Rubel, Brain Res. 1000:200-210, 2003 } } } } Regards, } } Glen } } On Jun 7, 2006, at 10:49 AM, drk-at-SHCC.org wrote: } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } } MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/ } } } FAQ.html } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } } } } Regarding the idea of localizing fluorescence label in epon, I } } } would suggest } } } that you not use Epon...there are autoflourescence problems (at } } } least in our } } } hands, using Spurrs resin). However, I think you may be happy } } } using LR } } } White, polymerized at 60C. We've done some similar work here and } } } have been } } } very impressed with the results. } } } } } } Good luck, } } } } } } Doug } } } } } } Douglas R. Keene } } } Assistant Investigator } } } Micro-Imaging Center } } } Shriners Hospital for Children } } } 3102 S.W. Sam Jackson Park Road } } } Portland, Oregon 97239 } } } 503-221-3434 } } } drk-at-shcc.org } } } -----Original Message----- } } } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } } } Sent: Wednesday, June 07, 2006 1:10 AM } } } To: drk-at-SHCC.org } } } Subject: [Microscopy] weird idea: fluorescence in Epon } } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } 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, } } } } } } Here is another weird idea from me :-D } } } I would like to localize a fluorescent molecule in the } } } transversal plane of a cell monolayer(not from above). } } } We have no material for the preparation of } } } histological sections, but we have all necessary for } } } TEM. I wonder if I could not embed the monolayer in } } } Epon, then cut transversal semi-thin sections and } } } observe in epifluroscence. } } } Would Epon hinder fluorescence? } } } Could it be done in another resin than Epon? } } } } } } Regards, } } } } } } StČphane } } }
==============================Original Headers============================== 9, 25 -- From glenmac-at-u.washington.edu Thu Jun 8 11:55:26 2006 9, 25 -- Received: from mxout1.cac.washington.edu (mxout1.cac.washington.edu [140.142.32.134]) 9, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58GtQZE014722 9, 25 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 11:55:26 -0500 9, 25 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.33.9]) 9, 25 -- by mxout1.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k58GtPsV019189 9, 25 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 09:55:25 -0700 9, 25 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 9, 25 -- (authenticated authid=glenmac) 9, 25 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k58GtMgj017005 9, 25 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 9, 25 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 09:55:25 -0700 9, 25 -- Mime-Version: 1.0 (Apple Message framework v750) 9, 25 -- In-Reply-To: {p06210229c0adfa70cf1a-at-[10.0.1.3]} 9, 25 -- References: {200606072039.k57KdsxX003708-at-ns.microscopy.com} {p06210229c0adfa70cf1a-at-[10.0.1.3]} 9, 25 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 25 -- Message-Id: {208F7BAE-D0C3-4CBF-B33A-BBFD3BDF8880-at-u.washington.edu} 9, 25 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 9, 25 -- Subject: RE: weird idea 9, 25 -- Date: Thu, 8 Jun 2006 09:55:17 -0700 9, 25 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 9, 25 -- X-Mailer: Apple Mail (2.750) 9, 25 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_NAME_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_FUNWORDS 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0' 9, 25 -- Content-Transfer-Encoding: 8bit 9, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k58GtQZE014722 ==============================End of - Headers==============================
Shrunali Could I suggest you check out the Microscopy List Server Archive? There has been extensive and intelligent discussion about this subject quite frequently over the last couple of years
Richard Harris Laboratory Supervisor, Imaging and Analysis Department of Biology, University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- X-from: aarti_harle-at-yahoo.co.in [mailto:aarti_harle-at-yahoo.co.in] Sent: June 8, 2006 7:48 AM To: rjharris-at-uwo.ca
Hello,
Can anyone suggest good scanner to scan TEM negatives and prints and and latest Imange Analysis software.
Regards Shrunali Kulkarni Scientist IMTech
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 4, 18 -- From aarti_harle-at-yahoo.co.in Thu Jun 8 06:42:41 2006 4, 18 -- Received: from web8324.mail.in.yahoo.com (web8324.mail.in.yahoo.com [202.43.219.162]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58BgdP1011677 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 06:42:40 -0500 4, 18 -- Received: (qmail 80614 invoked by uid 60001); 8 Jun 2006 11:42:38 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.co.in; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content -Transfer-Encoding; 4, 18 -- b=4O7v0G9DukweatO+2R8G5gg8q2vMGakVzH/e8sNmdfB5lHYsou5keT5GIf5MCJ3QXABL+p6TYN 2J8ErZVja7YfTq2GEVJSWlBmkVhK7a8GWj2EHuUnSD/dSKfIhsaVfnZmLPKTxxwkjJsKthJ4k7ai d29uMqt8F5DjHJQgxz+8E= ; 4, 18 -- Message-ID: {20060608114238.80612.qmail-at-web8324.mail.in.yahoo.com} 4, 18 -- Received: from [203.197.210.214] by web8324.mail.in.yahoo.com via HTTP; Thu, 08 Jun 2006 04:42:38 PDT 4, 18 -- Date: Thu, 8 Jun 2006 04:42:38 -0700 (PDT) 4, 18 -- From: Aarti Harle {aarti_harle-at-yahoo.co.in} 4, 18 --
You can purchase many different scanners for prices under $500 that will do the job for routine negatives. What is critical is that they are capable of both reflective and transmittal illumination. For negatives you need to be able to transmit the light through the negative while normal scanning of documents utilizes reflective light.
We normally scan routine negatives at 600dpi. However, if we need to enlarge the image substantially, we will often scan at 1200dpi or higher.
Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
} From: {mcauliff-at-umdnj.edu} } Reply-To: {mcauliff-at-umdnj.edu} } Date: Thu, 8 Jun 2006 08:37:52 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] Re: Scanner } } } } } ---------------------------------------------------------------------------- } 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 put my TEM negatives on a lightbox and photograph them with a Canon G3 } digital camera. Then I convert to a positive with PhotoShop. } } Geoff } } } aarti_harle-at-yahoo.co.in wrote: } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Hello, } } } } Can anyone suggest good scanner to scan TEM negatives } } and prints and and latest Imange Analysis software. } } } } Regards } } Shrunali Kulkarni } } Scientist } } IMTech } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam protection around } } http://mail.yahoo.com } } } } } } } } } -- } -- } ********************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane, Piscataway, NJ 08854 } voice: (732)-235-4583; fax: -4029 } mcauliff-at-umdnj.edu } ********************************************** } } } } ==============================Original Headers============================== } 9, 35 -- From mcauliff-at-umdnj.edu Thu Jun 8 08:35:24 2006 } 9, 35 -- Received: from zix04.umdnj.edu (zix04.UMDNJ.EDU [130.219.34.127]) } 9, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k58DZOFj011595 } 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:24 -0500 } 9, 35 -- Received: from zix04.umdnj.edu (ZixVPM [127.0.0.1]) } 9, 35 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id C348C4BE51 } 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:23 -0500 } (CDT) } 9, 35 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) } 9, 35 -- by zix04.umdnj.edu (Proprietary) with ESMTP id 05B9F4BE6A } 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:21 -0500 } (CDT) } 9, 35 -- Received: from ([130.219.34.131]) } 9, 35 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.16981519; } 9, 35 -- Thu, 08 Jun 2006 09:35:01 -0400 } 9, 35 -- Received: from conversion-daemon.Polaris.umdnj.edu by } Polaris.umdnj.edu } 9, 35 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) } 9, 35 -- id {0J0J00501N192Q-at-Polaris.umdnj.edu} (original mail from } mcauliff-at-umdnj.edu) } 9, 35 -- for microscopy-at-msa.microscopy.com; Thu, 08 Jun 2006 09:35:01 -0400 } (EDT) } 9, 35 -- Received: from [127.0.0.1] ([10.138.2.123]) } 9, 35 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 } (built Oct 21 } 9, 35 -- 2004)) with ESMTP id {0J0J005AJN211E-at-Polaris.umdnj.edu} ; Thu, } 9, 35 -- 08 Jun 2006 09:34:50 -0400 (EDT) } 9, 35 -- Date: Thu, 08 Jun 2006 09:35:48 -0400 } 9, 35 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} } 9, 35 -- Subject: Re: [Microscopy] Scanner } 9, 35 -- In-reply-to: {200606081144.k58BiOek013698-at-ns.microscopy.com} } 9, 35 -- To: aarti_harle-at-yahoo.co.in, } 9, 35 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} } 9, 35 -- Message-id: {448827B4.5000709-at-umdnj.edu} } 9, 35 -- MIME-version: 1.0 } 9, 35 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 } 9, 35 -- Content-transfer-encoding: 7BIT } 9, 35 -- X-Accept-Language: en-us, en } 9, 35 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) } 9, 35 -- Gecko/20040804 Netscape/7.2 (ax) } 9, 35 -- References: {200606081144.k58BiOek013698-at-ns.microscopy.com} } ==============================End of - Headers==============================
==============================Original Headers============================== 7, 23 -- From dsherman-at-purdue.edu Thu Jun 8 13:23:12 2006 7, 23 -- Received: from 1061exfe02.adpc.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58INBT5004486 7, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 13:23:12 -0500 7, 23 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe02.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 23 -- Thu, 8 Jun 2006 14:23:11 -0400 7, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 7, 23 -- Thu, 8 Jun 2006 18:23:08 +0000 7, 23 -- User-Agent: Microsoft-Entourage/11.2.3.060209 7, 23 -- Date: Thu, 08 Jun 2006 14:23:07 -0400 7, 23 -- Subject: Re: [Microscopy] Re: Scanner 7, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 23 -- To: {mcauliff-at-umdnj.edu} 7, 23 -- CC: "message to: MSA list" {microscopy-at-microscopy.com} 7, 23 -- Message-ID: {C0ADE34B.127D7%dsherman-at-purdue.edu} 7, 23 -- Thread-Topic: [Microscopy] Re: Scanner 7, 23 -- Thread-Index: AcaLKJa01VvqA/cbEdq0UwARJN08Mg== 7, 23 -- In-Reply-To: {200606081337.k58DbqJa015892-at-ns.microscopy.com} 7, 23 -- Mime-version: 1.0 7, 23 -- Content-type: text/plain; 7, 23 -- charset="US-ASCII" 7, 23 -- Content-transfer-encoding: 7bit 7, 23 -- X-OriginalArrivalTime: 08 Jun 2006 18:23:11.0835 (UTC) FILETIME=[99963AB0:01C68B28] ==============================End of - Headers==============================
Seeing at the Nanoscale IV, July 17-20, University of Pennsylvania, Philadelphia.
All users of SPM and metrology instruments are invited to Philadelphia next month to meet and discuss their work informally with colleagues and nanoscience pioneers from around the world!
The three-day, event-filled "Seeing at the Nanoscale" international conference includes two-and-a-half days of technical presentations, an evening poster session, and a reception at the National Constitution Center.
Technical Program: · Nanomechanical and Local Property Measurements · Visualization I: Biomolecules and Biological Processes · Visualization II: Materials and Polymer Systems · Measurements of Electrical, Optical, Magnetic, and Thermal Properties of Materials at the Nanoscale · Instrumentation: New Tools and Techniques for Nanoscience
For more information and to register online, please go to: www.veeco.com/nanoconference
____________________________ Marlene Carlyle Veeco Instruments Conference Coordinator 112 Robin Hill Road Santa Barbara, CA 93117 Tel: 805-967-1400 (ext. 2312) Fax: 805-967-7717 Email: mcarlyle-at-veeco.com ____________________________
==============================Original Headers============================== 8, 20 -- From MCarlyle-at-veeco.com Thu Jun 8 17:50:46 2006 8, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.VEECO.com [68.111.35.167]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58Mojq9021045 8, 20 -- for {Microscopy-at-Microscopy.com} ; Thu, 8 Jun 2006 17:50:45 -0500 8, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 20 -- content-class: urn:content-classes:message 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; 8, 20 -- charset="iso-8859-1" 8, 20 -- Subject: SPM - Seeing at the Nanoscale IV Conference 8, 20 -- Date: Thu, 8 Jun 2006 15:50:44 -0700 8, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F4201AD54D5-at-sboexch2.int.veeco.com} 8, 20 -- X-MS-Has-Attach: 8, 20 -- X-MS-TNEF-Correlator: 8, 20 -- Thread-Topic: SPM - Seeing at the Nanoscale IV Conference 8, 20 -- Thread-Index: AcaLTfmt6Y/+7q5oQh++fBJptUPOrw== 8, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 8, 20 -- To: {Microscopy-at-Microscopy.com} 8, 20 -- Content-Transfer-Encoding: 8bit 8, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k58Mojq9021045 ==============================End of - Headers==============================
Yes, this has been a recurring issue. The archives are a good source for reflection.
Also, one should scan as a transparency rather than as a negative. Then invert in PS or whatever image app you use. Maximum D value is highly desirable and this is engaged with transparencies.
gary g.
At 10:54 AM 6/8/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Thu Jun 8 18:06:35 2006 9, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58N6ZD1031643 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 18:06:35 -0500 9, 20 -- Received: (qmail 18405 invoked from network); 8 Jun 2006 16:06:34 -0700 9, 20 -- Received: by simscan 1.1.0 ppid: 18402, pid: 18403, t: 0.2173s 9, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 20 -- by qsmtp1 with SMTP; 8 Jun 2006 16:06:34 -0700 9, 20 -- Message-Id: {7.0.1.0.2.20060608160413.023955c8-at-gaugler.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 20 -- Date: Thu, 08 Jun 2006 16:06:35 -0700 9, 20 -- To: rjharris-at-uwo.ca 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] RE: Scanner 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200606081754.k58Hslnn029206-at-ns.microscopy.com} 9, 20 -- References: {200606081754.k58Hslnn029206-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both fisher.phyllis-at-mayo.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: I am considering adding a cathode luminescence tip to an old Centaurus backscatter detector on an Hitachi 4700 FESEM. Does anyone have experience with this tip and/or suggestions or comments regarding such an installation?
Dear Group, I am looking for someone who is going to participate the meeting on Microscopy and Microanalysis 2006 in Chicago and wants to share a room (hotel, hostel or guest house). Please inform me offline!
Grzegorz Tylko Department of Cytology and Histology Institute of Zoology Jagiellonian Univeristy Ingardena 6, 30-060 Krakow Poland tel.: +48 12 663 24 25 fax: +48 12 634 49 51 mobile: +48 698 55 55 85 e-mail: tylko-at-zuk.iz.uj.edu.pl
==============================Original Headers============================== 4, 37 -- From tylko-at-zuk.iz.uj.edu.pl Fri Jun 9 01:45:34 2006 4, 37 -- Received: from theta.uoks.uj.edu.pl (theta.uoks.uj.edu.pl [149.156.89.30]) 4, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k596jXRs028765 4, 37 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 01:45:33 -0500 4, 37 -- Received: from zuk.iz.uj.edu.pl (zuk.iz.uj.edu.pl [149.156.72.18]) 4, 37 -- by theta.uoks.uj.edu.pl (8.13.6/8.13.6) with ESMTP id k596jUlp002534 4, 37 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 08:45:31 +0200 (CEST) 4, 37 -- Received: from ZUK/SpoolDir by zuk.iz.uj.edu.pl (Mercury 1.48); 4, 37 -- 9 Jun 06 08:45:31 +0100 4, 37 -- Received: from SpoolDir by ZUK (Mercury 1.48); 9 Jun 06 08:45:08 +0100 4, 37 -- Received: from gt (149.156.77.111) by zuk.iz.uj.edu.pl (Mercury 1.48); 4, 37 -- 9 Jun 06 08:45:04 +0100 4, 37 -- Message-ID: {003301c68b90$3d231a60$6f4d9c95-at-gt} 4, 37 -- From: "Grzegorz Tylko" {tylko-at-zuk.iz.uj.edu.pl} 4, 37 -- To: {microscopy-at-microscopy.com} 4, 37 -- Subject: MM2006-accommodation 4, 37 -- Date: Fri, 9 Jun 2006 08:45:04 +0200 4, 37 -- MIME-Version: 1.0 4, 37 -- Content-Type: text/plain; 4, 37 -- format=flowed; 4, 37 -- charset="iso-8859-2"; 4, 37 -- reply-type=original 4, 37 -- Content-Transfer-Encoding: 7bit 4, 37 -- X-Priority: 3 4, 37 -- X-MSMail-Priority: Normal 4, 37 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 4, 37 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 4, 37 -- X-Antivirus: Dr.Web (R) for Mail Servers on pandora host 4, 37 -- X-Antivirus-Code: 100000 4, 37 -- X-Spam-Flag: NO 4, 37 -- X-Spam-Status: score=-5.9 required=5.0 4, 37 -- X-Spam-Report: 4, 37 -- * -3.3 ALL_TRUSTED Passed through trusted hosts only via SMTP 4, 37 -- * -2.6 BAYES_00 BODY: Bayesian spam probability is 0 to 1% 4, 37 -- * [score: 0.0000] 4, 37 -- X-Spam-Checker-Version: SpamAssassin 3.0.5 (2005-11-28) on 4, 37 -- pandora.uoks.uj.edu.pl ==============================End of - Headers==============================
The only problem with archived scanner info is that it goes out of date the minute a new scanner hits the streets. I bought a Ł250 Canon 9550F at home [Xmas 2005] under the mistaken view that consumer flatbeds can't get any better than this for the price - I was very wrong (and annoyingly it was my money). Going back 5 years a scanner that produced images like the 9950F would have cost Ł4,000.
And I remember paying Ł5,000 for an office 386 computer with 8Mb of RAM and a 120Mb hard drive back in 1989 - I wouldn't recommend it as particularly good value these days. It did allow me to ditch the expensive IBM central mainframe and run Fortran FTN77 independently on my PC though.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: rjharris-at-uwo.ca [mailto:rjharris-at-uwo.ca] Sent: 08 June 2006 18:57 To: keith.morris-at-ucl.ac.uk
Shrunali Could I suggest you check out the Microscopy List Server Archive? There has been extensive and intelligent discussion about this subject quite frequently over the last couple of years
Richard Harris Laboratory Supervisor, Imaging and Analysis Department of Biology, University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- X-from: aarti_harle-at-yahoo.co.in [mailto:aarti_harle-at-yahoo.co.in] Sent: June 8, 2006 7:48 AM To: rjharris-at-uwo.ca
Hello,
Can anyone suggest good scanner to scan TEM negatives and prints and and latest Imange Analysis software.
Regards Shrunali Kulkarni Scientist IMTech
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
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Hi all,
I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated.
Thank You,
Jennifer Ray Sloppy Materials Science & Engineering Department Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 9, 17 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 11:45:10 -0500 9, 17 -- Received: (from webmail-at-localhost) 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Message-Id: {200606091645.MAA01431-at-webmail15.cac.psu.edu} 9, 17 -- To: microscopy-at-microscopy.com 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 9, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 9, 17 -- X-Mailer: Penn State WebMail 9, 17 -- X-Sender: jds451 9, 17 -- X-Originating-IP: 128.118.37.54 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
Jennifer Ray Sloppy wrote: ==================================================== I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated. ============================================================ Some number of years ago, when our firm was first getting active in surface analysis and the need for calibrating etch rates became apparent, and the "standard" at the time for calibrating etching rates was to etch through an anodic film grown on a thin tantalum foil of known thickness, we found this could be cross-sectioned by diamond knife ultramicrotomy. Other than possessing a lot of experience with the ultramicrotomy of "difficult and hard" samples, we vacuum embedded the sample with our own SPI-Pon™ 812 epoxy embedding resin, and did the thin sectioning with a 45 deg angle SPI Supplies® Brand Materials Science Diamond Knife but with a fairly ordinary (e.g. not very new) ultramicrotome. Separation of the anodized layer from the embedding resin could be minimized by using a vacuum impregnation. We were not always successful in keeping the anodized layer/substrate tantalum interface intact. We could get less separation by using a 35 deg knife but this wore out the knife very quickly and since we did not mind some separation, we used mainly a 45 deg. knife.
But one could get good TEM views of the ultramicrotome-produced cross-section for making (anodized layer) thickness measurements and high resolution images showing the morphology and fine structure of the anodized layer.
Disclaimer: SPI Supplies offers both the above-mentioned embedding resin and diamond knife and we also offer, through Structure Probe, Inc. this kind of a laboratory service for others.
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============================== 16, 24 -- From cgarber-at-2spi.com Fri Jun 9 13:43:39 2006 16, 24 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59IhdBg009769 16, 24 -- for {microscopy-at-msa.microscopy.com} ; Fri, 9 Jun 2006 13:43:39 -0500 16, 24 -- Received: from ibm1x23g2abfyg ([68.246.87.223]) 16, 24 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k59IhXNu008521; 16, 24 -- Fri, 9 Jun 2006 14:43:36 -0400 16, 24 -- X-IDV-FirstRcvd: [68.246.87.223] 16, 24 -- X-IDV-HELO: ibm1x23g2abfyg 16, 24 -- Message-ID: {00b401c68bf4$9b4db500$df57f644-at-ibm1x23g2abfyg} 16, 24 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 16, 24 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 16, 24 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 16, 24 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 16, 24 -- Date: Fri, 9 Jun 2006 14:39:43 -0400 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-Type: text/plain; 16, 24 -- charset="Windows-1252" 16, 24 -- X-Priority: 3 16, 24 -- X-MSMail-Priority: Normal 16, 24 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 16, 24 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 16, 24 -- Content-Transfer-Encoding: 8bit 16, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k59IhdBg009769 ==============================End of - Headers==============================
I have a colleague who wants to do in situ hybridizations on sponge larvae (yes, sponges are animals and they have larval forms!). The larvae are less than a millimeter long and quite delicate.
She tried fixing some in 4% paraformaldehyde in sea water, and the larvae immediately exploded. She also tried Carnoy's fix, and 4% PFA in Millonig's phosphate buffer, and the same thing happened.
The only thing I could think of to suggest was dropping to 1 or 2% PFA in sea water. I've seen references to using Bouins for light microscopy or osmium tetroxide for TEM, but I don't think either of these approaches would be good for in situs.
Any other suggestions?
Gary P. Radice Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173
==============================Original Headers============================== 9, 21 -- From gradice-at-richmond.edu Fri Jun 9 13:48:42 2006 9, 21 -- Received: from monty.richmond.edu (monty.richmond.edu [141.166.24.29]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59Imfs9015519 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 13:48:42 -0500 9, 21 -- Received: from rayon.richmond.edu (rayon.richmond.edu [141.166.30.10]) 9, 21 -- by monty.richmond.edu (8.11.6/8.11.6) with ESMTP id k59Imf119051 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 14:48:41 -0400 9, 21 -- Received: from [141.166.185.175] (ffa185175.richmond.edu [141.166.185.175]) 9, 21 -- by rayon.richmond.edu (8.12.11.20060308/8.12.11) with ESMTP id k59ImfT5023124 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 14:48:41 -0400 9, 21 -- Mime-Version: 1.0 (Apple Message framework v750) 9, 21 -- Content-Transfer-Encoding: 7bit 9, 21 -- Message-Id: {063B77D0-FAF3-4B5D-AB90-04354229FE6A-at-richmond.edu} 9, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 21 -- To: microscopy-at-microscopy.com 9, 21 -- From: Gary Radice {gradice-at-richmond.edu} 9, 21 -- Subject: fixing sponge larvae for in situ hybridization 9, 21 -- Date: Fri, 9 Jun 2006 14:47:52 -0400 9, 21 -- X-Mailer: Apple Mail (2.750) 9, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED 9, 21 -- X-Scanned-By: MIMEDefang 2.49 on 141.166.30.10 ==============================End of - Headers==============================
There is a trick that I pulled with MoS2 films on Si in which I had similar problems as you are having. What I did was to first deposit the film through a gridded mask (I think that it was 400 mesh, but can't remember for sure.) Then I prepared the samples as normal. What happens is that you have enough Si-epoxy-Si areas when the samples are put together to hold the sample stack together. It worked quite well. For you, you will have to make a mask to prevent the anodic Ta2O5 from forming and then remove your mask prior to making your cross section.
I also have another solution that might work for you. These are Stainless steel rods that were electrospark cut with slits in them. You must precisely cut the width of two strips of material and remove the burrs and put them together between the rods. This gives you more support with less glue interfaces. If you send me your address, I can send you some 3mm tubes and some rods to try out.
The third method that might work for you is the Technoorg-Linda method. This is illustrated in the Arpad Barna article: Barna, A. (1992) "Topographic kinetics and practice of low-angle ion beam thinning", Specimen Preparation for TEM of Materials. III, Mat. Res. Soc. Symp. Proc. Vol. 254. R. Anderson, B. Tracy, and J. Bravman, eds. pp. 3–22. You can also check the following articles: "Preparation of Cross-Sectional TEM Samples for Low-Angle Ion Milling", J.P. MCCAFFREY AND A. BARNA, MICROSCOPY RESEARCH AND TECHNIQUE 36:362–367 (1997). I can also send you a copy of a video that shows how to do it. Basically, two pieces of the sample are cut down to precise sizes and put in a special titanium grid face-to-face. Then the grid is manipulated with tools to clamp the samples in place tightly. The assembly is then put into an epoxy. You can use M-bond 610, but T-L recommends a special Araldit mixture with carbon that avoids the differential sputter removal problem with non-filled epoxies. The sample is then ground down with 600 grit paper to a thickness of about 30-50 µm. You polish with 1 µm paste to finish. It is then ion milled at a high voltage and a low angle, say about 5 to 7 degrees with full rotation for say about an hour depending on the sputter rate. This will give a dimple in the epoxy joint. Then go to a very low angle, ~2 degrees, on one side of the sample and mill until the topography is pushed to one side of the sample. Not all commercially available ion mills are capable of going to such a low angle. You must use a holder that is capable of it. Some mills have holders that are more massive that allow the heat to be dissipated more readily. Use the holder that is made to be milled from one side and at very low angles. You will see the topography of the sample pushed to the opposite side of the sample from where the gun is. Turn the sample over and mill in such a way that the ion direction would appear to oppose the direction from the first side milling. When you are done, the topography will again be pushed away from the ion gun and it will be on the opposite side of the sample of the glue joint than where it is on the first side. Finish the sample using the same angles, but low energy. Unless you have a special low energy gun such as the one available from Technoorg-Linda, most commercially available guns are limited to about 3 kV because they do not efficiently ionize the gas and the beam spread. The low energy guns available from T-L will go as low as 100 eV.
If you have any questions, please give me a call.
-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: jds451-at-psu.edu [mailto:jds451-at-psu.edu] Sent: Friday, June 09, 2006 9:49 AM To: Walck-at-SouthBayTech.com
Hi all,
I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated.
Thank You,
Jennifer Ray Sloppy Materials Science & Engineering Department Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 9, 17 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 11:45:10 -0500 9, 17 -- Received: (from webmail-at-localhost) 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Message-Id: {200606091645.MAA01431-at-webmail15.cac.psu.edu} 9, 17 -- To: microscopy-at-microscopy.com 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 9, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 9, 17 -- X-Mailer: Penn State WebMail 9, 17 -- X-Sender: jds451 9, 17 -- X-Originating-IP: 128.118.37.54 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
==============================Original Headers============================== 24, 28 -- From walck-at-southbaytech.com Fri Jun 9 14:21:22 2006 24, 28 -- Received: from ylpvm29.prodigy.net (ylpvm29-ext.prodigy.net [207.115.57.60]) 24, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59JLLic030645 24, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 14:21:21 -0500 24, 28 -- Received: from pimout5-ext.prodigy.net (pimout5-int.prodigy.net [207.115.4.21]) 24, 28 -- by ylpvm29.prodigy.net (8.12.10 outbound/8.12.10) with ESMTP id k59JKHVu031016 24, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 15:20:18 -0400 24, 28 -- X-ORBL: [64.169.193.90] 24, 28 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 24, 28 -- by pimout5-ext.prodigy.net (8.13.6 out.dk/8.13.6) with ESMTP id k59JLG3o212652; 24, 28 -- Fri, 9 Jun 2006 15:21:16 -0400 24, 28 -- From: "Scott Walck" {walck-at-southbaytech.com} 24, 28 -- To: {jds451-at-psu.edu} 24, 28 -- Cc: {Microscopy-at-microscopy.com} 24, 28 -- Subject: RE: [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep 24, 28 -- Date: Fri, 9 Jun 2006 12:21:21 -0700 24, 28 -- Message-ID: {002801c68bf9$e4fd4940$7801a8c0-at-dynamicbl8uno3} 24, 28 -- MIME-Version: 1.0 24, 28 -- Content-Type: text/plain; 24, 28 -- charset="iso-8859-1" 24, 28 -- X-Priority: 3 (Normal) 24, 28 -- X-MSMail-Priority: Normal 24, 28 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 24, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, 28 -- Importance: Normal 24, 28 -- In-Reply-To: {200606091648.k59GmbKg003550-at-ns.microscopy.com} 24, 28 -- Content-Transfer-Encoding: 8bit 24, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k59JLLic030645 ==============================End of - Headers==============================
For cross sectional samples of metal/metal oxide, a rocking motion in ion mill (or single section mode in PIPS) serves best. Align the glue line perpendicular with the ion beam, allowing the beam ONLY bombarding from the backside of the film. With such arrangement, the substrate is acting as a shield for the oxide film, protecting it from being overly milled.
Chengyu Song National Center for Electron Microscopy Lawrence Berkeley National Laboratory
jds451-at-psu.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 been attempting to prepare cross sectional TEM samples of anodic tantala } (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an } approximate thickness of 200um. The oxide film is grown anodically on the Ta } using electrochemistry. } } In the literature, I have seen cross sectional TEM images of anodic tantala } grown on sputter deposited tantalum; an aluminum or silicon substrate is } typically used. I am curious if anyone has had success in preparing anodic } Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer) } } My main problem seems to be with de-lamination of the cross section sandwich. } Here is my attempted procedure: } } I first bound two faces of my sample together and placed the sandwich in a press } while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich } frequently breaks apart when I begin polishing. To address this problem, I } encased my sandwich in a brass tube and fitted the tube with supporting brass } shims, all held in place with epoxy. Using this technique, I was able to thin } some parts of the specimen to 20-50 um, but the epoxy was no longer present at } this point. When I attempted to ion-mill, the sandwiches splintered apart, and } any of the oxide that may have been surviving was obliterated in the ion mill. } } Any suggestions or advice would be greatly appreciated. } } Thank You, } } Jennifer Ray Sloppy } Materials Science & Engineering Department } Ph.D. Candidate } Pennsylvania State University } } } ==============================Original Headers============================== } 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 } 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) } 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 } 9, 17 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 11:45:10 -0500 } 9, 17 -- Received: (from webmail-at-localhost) } 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; } 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) } 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) } 9, 17 -- Message-Id: {200606091645.MAA01431-at-webmail15.cac.psu.edu} } 9, 17 -- To: microscopy-at-microscopy.com } 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep } 9, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} } 9, 17 -- X-Mailer: Penn State WebMail } 9, 17 -- X-Sender: jds451 } 9, 17 -- X-Originating-IP: 128.118.37.54 } 9, 17 -- MIME-Version: 1.0 } 9, 17 -- Content-Type: text/plain } ==============================End of - Headers==============================
==============================Original Headers============================== 6, 25 -- From csong-at-lbl.gov Fri Jun 9 17:56:04 2006 6, 25 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 6, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59Mu3DI012665 6, 25 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 17:56:03 -0500 6, 25 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 6, 25 -- by mta2.lbl.gov (8.13.6/8.13.6) with ESMTP id k59Mu2tk022358 6, 25 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 15:56:02 -0700 (PDT) 6, 25 -- Received: from lbl.gov (apple-0-5-2-e0-6a-f3.dhcp.lbl.gov [131.243.3.239]) 6, 25 -- by mta2.lbl.gov (8.13.6/8.13.6) with ESMTP id k59Mu1Ys022355; 6, 25 -- Fri, 9 Jun 2006 15:56:01 -0700 (PDT) 6, 25 -- Message-ID: {4489FCBD.6117147D-at-lbl.gov} 6, 25 -- Date: Fri, 09 Jun 2006 15:57:49 -0700 6, 25 -- From: chengyu song {csong-at-lbl.gov} 6, 25 -- Reply-To: csong-at-lbl.gov 6, 25 -- Organization: lbl 6, 25 -- X-Mailer: Mozilla 4.73C-CCK-MCD LBNL V4.73 Build 2 (Macintosh; U; PPC) 6, 25 -- X-Accept-Language: en,pdf 6, 25 -- MIME-Version: 1.0 6, 25 -- To: jds451-at-psu.edu, microscopy-at-microscopy.com 6, 25 -- Subject: Re: [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep 6, 25 -- References: {200606091648.k59GmVj4003324-at-ns.microscopy.com} 6, 25 -- Content-Type: text/plain; charset=gb2312; x-mac-type="54455854"; x-mac-creator="4D4F5353" 6, 25 -- Content-Transfer-Encoding: 7bit 6, 25 -- X-Virus-Scanned: ClamAV 0.88.2/1524/Fri Jun 9 14:28:03 2006 on mta2 6, 25 -- X-Virus-Status: Clean ==============================End of - Headers==============================
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Title-Subject: [Filtered] SEM - cathode luminescence detector
Question: I am considering adding a cathode luminescence tip to an old Centaurus backscatter detector on an Hitachi 4700 FESEM. Do you have any suggestions or comments on such a move?
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Email: warner.rr-at-pg.com Name: Ron Warner
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Title-Subject: [Filtered] Reichert CS Auto, free to a good home
Question: We have the ancient Reichert CS Auto freeze substitution unit available to a good home. It was working when we last used it, about 3 years ago. The body shows some of its years (so does mine), but the internals should be fine. You pay the shipping.
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Email: aetmicro-at-optonline.net Name: Andrew Thelian
Organization: Nanoprobes, Inc.
Title-Subject: [Filtered] Philips EM 300 camera chamber issues
Question: Hi All :),
I had an issues with the plate chambers on my Philips EM 300... The venting valve jammed... So I naturally took it upon myself to attempt a repair... the overall repair seemed to have worked, the valve knob no longer jams or sticks like it had...
my problem now is...
when turning the knob(clockwise) to 10/4 o clock... the vacuum pump initiates like it is suppose to... i then press the knob in creating a seal(i wait on the vacuum light sequence)... then the last step is to finish the circuit and leave the knob at its resting position at 9/3 o clock... the rough pump shuts off as i attempt to complete the circuit...
I really doubt that PFA in itseld could make a structure explode. Your colleague should look at the osmoticity of the medium used for the fixation. I don't know sponges very well, but there is a chance that the intracellular osmoticity is regulated and is not the same as the extracellular medium (sea water). If the fixative stops the ion pumps present in the membrane, it may be possible that the high salt concentration of sea water provokes an osmotic shock (even though in this case one should expect a shrinkage of the cell, but you never know). If the intracellular osmotocity of sponge cells is unknown, he'd better try fixation media with different osmotic forces.
Stephane
--- gradice-at-richmond.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a colleague who wants to do in situ } hybridizations on sponge } larvae (yes, sponges are animals and they have } larval forms!). The } larvae are less than a millimeter long and quite } delicate. } } } She tried fixing some in 4% paraformaldehyde in sea } water, and the } larvae immediately exploded. She also tried Carnoy's } fix, and 4% PFA } in Millonig's } phosphate buffer, and the same thing happened. } } The only thing I could think of to suggest was } dropping to 1 or 2% } PFA in sea water. I've seen references to using } Bouins for light } microscopy or osmium tetroxide for TEM, but I don't } think either of } these approaches would be good for in situs. } } } Any other suggestions? } } } Gary P. Radice } Department of Biology 804-289-8107 (voice) } University of Richmond 804-289-8233 (FAX) } Richmond VA 23173 } } } ==============================Original } Headers============================== } 9, 21 -- From gradice-at-richmond.edu Fri Jun 9 } 13:48:42 2006 } 9, 21 -- Received: from monty.richmond.edu } (monty.richmond.edu [141.166.24.29]) } 9, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k59Imfs9015519 } 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 } Jun 2006 13:48:42 -0500 } 9, 21 -- Received: from rayon.richmond.edu } (rayon.richmond.edu [141.166.30.10]) } 9, 21 -- by monty.richmond.edu (8.11.6/8.11.6) with } ESMTP id k59Imf119051 } 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 } Jun 2006 14:48:41 -0400 } 9, 21 -- Received: from [141.166.185.175] } (ffa185175.richmond.edu [141.166.185.175]) } 9, 21 -- by rayon.richmond.edu } (8.12.11.20060308/8.12.11) with ESMTP id } k59ImfT5023124 } 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 } Jun 2006 14:48:41 -0400 } 9, 21 -- Mime-Version: 1.0 (Apple Message framework } v750) } 9, 21 -- Content-Transfer-Encoding: 7bit } 9, 21 -- Message-Id: } {063B77D0-FAF3-4B5D-AB90-04354229FE6A-at-richmond.edu} } 9, 21 -- Content-Type: text/plain; charset=US-ASCII; } delsp=yes; format=flowed } 9, 21 -- To: microscopy-at-microscopy.com } 9, 21 -- From: Gary Radice {gradice-at-richmond.edu} } 9, 21 -- Subject: fixing sponge larvae for in situ } hybridization } 9, 21 -- Date: Fri, 9 Jun 2006 14:47:52 -0400 } 9, 21 -- X-Mailer: Apple Mail (2.750) } 9, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED } 9, 21 -- X-Scanned-By: MIMEDefang 2.49 on } 141.166.30.10 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 8, 20 -- From nizets2-at-yahoo.com Mon Jun 12 02:48:31 2006 8, 20 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5C7mVOh008885 8, 20 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 02:48:31 -0500 8, 20 -- Received: (qmail 1895 invoked by uid 60001); 12 Jun 2006 07:48:30 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=aQZxRVgMXGOewWw84RPkTRum1TKUECd1jKiE4vTgiSiUlxyZU6MXAexB+h8c1oO9fBMN2EDCBtGE2krNmPSuTCm4032fu7lSkwMC39aWgnfANNEbEv7I6ZUoKxkaTk14YiSdlp2eZqP8YPWj7g0CxPiCufoTJy7eFTBIm1rVQr4= ; 8, 20 -- Message-ID: {20060612074830.1893.qmail-at-web37410.mail.mud.yahoo.com} 8, 20 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Mon, 12 Jun 2006 00:48:30 PDT 8, 20 -- Date: Mon, 12 Jun 2006 00:48:30 -0700 (PDT) 8, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 20 -- Subject: Re: [Microscopy] fixing sponge larvae for in situ hybridization 8, 20 -- To: gradice-at-richmond.edu 8, 20 -- Cc: microscopy-at-microscopy.com 8, 20 -- In-Reply-To: {200606091852.k59Iqpon027420-at-ns.microscopy.com} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I notice that the Epson V750 Pro flatbed scanner now has an optional fluid mounting accessory that involves a bottom sealed lift out tray (that 'prevents' fluid seepage into the scanner and is 'easy clean'). See it at:
Professional drum scanners use mounting fluid to improve film scan quality if you really need the best possible scan, and have a private income (most of us don't). If you are flatbed scanning odd specimens or really want to use oil or mounting fluid, this accessory looks very useful. I don't think the kit's generally available yet though.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: mey-at-amnh.org [mailto:mey-at-amnh.org] Sent: 25 May 2006 20:47 To: keith.morris-at-ucl.ac.uk
Hi Mike, I have been exploring and having success using a flatbed scanner for particle analysis, but actually started out in this endeavor by scanning rocks and rock cores. We now use an Epson XL 1640 scanner, which has a focus function, essentially allowing us to get right on the surface of the samples. However, a pretty flat cut is needed to get overall excellent quality. I found that this works much better than greasing up the bed with oils, glycerin or other sticky stuff. We used a thin plastic sheet (same material as thin clear view-foils) to protect the glass from the rocks. As a side note, I too saw some strange artifacts with oils, which could (?) be due to "internal" reflections or interference patterns between the glass-plastic or micro-bubbles on or close the sample surface. My major concern was contaminating the samples with oils, especially when doing trace element work or isotope analysis for age dating purposes of the rock cores. Not that the oil itself would be the major culprit, but more the rock dust flying around everywhere in the cutting lab sticking to the greasy surfaces. Rock cores sometimes have oil on them from the drilling process and is usually washed off with sulfo. However, the porosity of the sample may contribute to deeper contamination, especially is if the sample shows cracks or micro-fractures. I had to consider this for archival purposes and potential future analysis on the same samples as new technologies emerge.
michael-at-Shaffer.net wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } We have been using a flatbed scanner for documenting rock core before } processing it for other analytical techniques. Now, we want to explore the } technique a bit further, possibly towards applied image analysis, but need } one last ingredient for better quality scans. We want to use some akin to } immersion oil for the interface between rock and flatbed, but want to find } something inexpensive, non-toxic and easy to clean. Any thoughts? } } TIA & 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 } } } } } ==============================Original Headers============================== } 7, 19 -- From Michael-at-Shaffer.net Thu May 25 05:52:02 2006 } 7, 19 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4PAq1qK008574 } 7, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 25 May 2006 05:52:01 -0500 } 7, 19 -- Received: (qmail 17426 invoked from network); 25 May 2006 10:52:00 -0000 } 7, 19 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) } 7, 19 -- by ws6-4.us4.outblaze.com with SMTP; 25 May 2006 10:52:00 -0000 } 7, 19 -- From: "michael shaffer" {michael-at-Shaffer.net} } 7, 19 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} } 7, 19 -- Subject: imaging rock core with flatbed scanner } 7, 19 -- Date: Thu, 25 May 2006 08:21:59 -0230 } 7, 19 -- Message-ID: {000f01c67fe9$40c556e0$8d829986-at-roamingwolf} } 7, 19 -- MIME-Version: 1.0 } 7, 19 -- Content-Type: text/plain; } 7, 19 -- charset="US-ASCII" } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- X-Mailer: Microsoft Office Outlook 11 } 7, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 7, 19 -- Thread-Index: AcZ/6T9zCvUQjGd1R8+CGbe9ZHz2pA== } ==============================End of - Headers============================== } }
Greetings listers. We have an image analysis problem, and I could use your help. We are frequently asked to characterize the cell size and structure of various polymer foams for our customers. To date we have relied upon an ASTM method that involves counting the number of cell intersections with a reference line drawn on the image to compute average cell size. The method assumes a degree of homogeneity and cell density that is often an imperfect representation of our customer's samples, and in any event it would be far more desirable to select and measure the cells directly with our ImageProPlus software. There's the rub. We're having the devil of a time getting good segmentation of the open cells against the background matrix. We've tried a number of filters and operations, but thus far without success. I have posted two examples through the link listed below (I disabled it to get through the spam filter). To download, just right click on the file name and "save target as".
**http://www.impactanalytical.com/data/**
If any of you experts out there can set us on the right track, or alternatively explain why we're wasting our time, I would GREATLY appreciate it. My head hurts and the walls are in need of repair!
Incidentally, I have tried varying kV, and collecting in backscatter as well as backscatter plus/minus for topography. Thus far these efforts have not solved the fundamental problem that contrast along the rim of a given cell shifts from light to dark, confounding segmentation. Thanks for your consideration - there's a windy city beer in the bargain if my travel request goes through! Yours, Matt Matthew Stephenson Impact Analytical/MMI 1910 W. Saint Andrews Rd. Midland, MI 48640 (989)832-5555 X506
==============================Original Headers============================== 1, 14 -- From stephenson-at-impactanalytical.com Mon Jun 12 07:19:49 2006 1, 14 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CCJnEA003629 1, 14 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Jun 2006 07:19:49 -0500 1, 14 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 1, 14 -- id {M30M2R2Q} ; Mon, 12 Jun 2006 08:19:47 -0400 1, 14 -- Message-ID: {089C7709BE9235448E3622C6F38D828F06C69196-at-mitconexch.mitcon.org} 1, 14 -- From: "Stephenson, Matthew" {stephenson-at-impactanalytical.com} 1, 14 -- To: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 1, 14 -- Subject: Image Analysis problem 1, 14 -- Date: Mon, 12 Jun 2006 08:19:46 -0400 1, 14 -- MIME-Version: 1.0 1, 14 -- X-Mailer: Internet Mail Service (5.5.2656.59) 1, 14 -- Content-Type: text/plain ==============================End of - Headers==============================
Your images are nearly identical to one that I use as a lab experiment in the image analysis courses I teach.
If you examine the image visually, how do you "see" the edges of the pores? There is always some local brightness change associated with the edge. A good way to highlight these edges is to duplicate the image, and perform a morphological erosion to replace each pixel with its darkest neighbor. Then subtract that image from the original, which will leave the locallly lightest pixels. These highlight the edges. Of course, you will want to do a bit of cleanup on the images first (a median filter or one of its derivatives is probably better than a smoothing convolution).
Your major problem is not ** measuring ** the circular features in your image, it is relating those measurements to the pores. This is not an imaging problem but a stereological one. The sections through the pores are NOT the pore sizes. There is a lot of literature on the relationships between the sizes of 2D sections through features and the sizes of the 3D structures.
John Russ
-----Original Message----- X-from: stephenson-at-impactanalytical.com To: DrJohnRuss-at-AOL.com Sent: Mon, 12 Jun 2006 07:20:35 -0500
Greetings listers. We have an image analysis problem, and I could use your help. We are frequently asked to characterize the cell size and structure of various polymer foams for our customers. To date we have relied upon an ASTM method that involves counting the number of cell intersections with a reference line drawn on the image to compute average cell size. The method assumes a degree of homogeneity and cell density that is often an imperfect representation of our customer's samples, and in any event it would be far more desirable to select and measure the cells directly with our ImageProPlus software. There's the rub. We're having the devil of a time getting good segmentation of the open cells against the background matrix. We've tried a number of filters and operations, but thus far without success. I have posted two examples through the link listed below (I disabled it to get through the spam filter). To download, just right click on the file name and "save target as".
**http://www.impactanalytical.com/data/**
If any of you experts out there can set us on the right track, or alternatively explain why we're wasting our time, I would GREATLY appreciate it. My head hurts and the walls are in need of repair!
Incidentally, I have tried varying kV, and collecting in backscatter as well as backscatter plus/minus for topography. Thus far these efforts have not solved the fundamental problem that contrast along the rim of a given cell shifts from light to dark, confounding segmentation. Thanks for your consideration - there's a windy city beer in the bargain if my travel request goes through! Yours, Matt Matthew Stephenson Impact Analytical/MMI 1910 W. Saint Andrews Rd. Midland, MI 48640 (989)832-5555 X506
==============================Original Headers============================== 1, 14 -- From stephenson-at-impactanalytical.com Mon Jun 12 07:19:49 2006 1, 14 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CCJnEA003629 1, 14 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Jun 2006 07:19:49 -0500 1, 14 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 1, 14 -- id {M30M2R2Q} ; Mon, 12 Jun 2006 08:19:47 -0400 1, 14 -- Message-ID: {089C7709BE9235448E3622C6F38D828F06C69196-at-mitconexch.mitcon.org} 1, 14 -- From: "Stephenson, Matthew" {stephenson-at-impactanalytical.com} 1, 14 -- To: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 1, 14 -- Subject: Image Analysis problem 1, 14 -- Date: Mon, 12 Jun 2006 08:19:46 -0400 1, 14 -- MIME-Version: 1.0 1, 14 -- X-Mailer: Internet Mail Service (5.5.2656.59) 1, 14 -- Content-Type: text/plain ==============================End of - Headers==============================
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==============================Original Headers============================== 17, 25 -- From DrJohnRuss-at-aol.com Mon Jun 12 08:09:54 2006 17, 25 -- Received: from imo-m20.mx.aol.com (imo-m20.mx.aol.com [64.12.137.1]) 17, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CD9r0E014603 17, 25 -- for {microscopy-at-sparc5.microscopy.com} ; Mon, 12 Jun 2006 08:09:53 -0500 17, 25 -- Received: from DrJohnRuss-at-aol.com 17, 25 -- by imo-m20.mx.aol.com (mail_out_v38_r7.5.) id v.4c9.f9cb4e (15886); 17, 25 -- Mon, 12 Jun 2006 09:09:47 -0400 (EDT) 17, 25 -- Received: from MBLK-M40 (mblk-m40.mblk.aol.com [64.12.136.84]) by air-id08.mx.aol.com (v109.13) with ESMTP id MAILINID81-3e0e448d679b241; Mon, 12 Jun 2006 09:09:47 -0400 17, 25 -- Content-Transfer-Encoding: 7bit 17, 25 -- Date: Mon, 12 Jun 2006 09:09:47 -0400 17, 25 -- Message-Id: {8C85C389D78B718-17B0-4362-at-MBLK-M40.sysops.aol.com} 17, 25 -- From: drjohnruss-at-aol.com 17, 25 -- References: {200606121220.k5CCKZt8004417-at-ns.microscopy.com} 17, 25 -- Cc: stephenson-at-impactanalytical.com 17, 25 -- Received: from 65.190.140.228 by MBLK-M40.sysops.aol.com (64.12.136.84) with HTTP (WebMailUI); Mon, 12 Jun 2006 09:09:47 -0400 17, 25 -- X-MB-Message-Source: WebUI 17, 25 -- X-MB-Message-Type: User 17, 25 -- In-Reply-To: {200606121220.k5CCKZt8004417-at-ns.microscopy.com} 17, 25 -- X-Mailer: AOL WebMail 17789 17, 25 -- Subject: Re: [Microscopy] Image Analysis problem 17, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 17, 25 -- MIME-Version: 1.0 17, 25 -- To: microscopy-at-ns.microscopy.com 17, 25 -- X-AOL-IP: 64.12.136.84 17, 25 -- X-Spam-Flag: NO ==============================End of - Headers==============================
There have been several good suggestions already, so I will try to add a few hints that have worked for me. First of all, do make sure you are not
milling in the same direction as the epoxy line, as that will preferentially mill the epoxy away. Use beam modulation or the equivalent
to restrict the beam to milling perpendicular to the interface. Also, I've noticed the thinner the epoxy line, the better the milling. When gluing your two sample pieces together, make sure the surfaces are absolutely clean, check the epoxy (I always use G-1) for specks of 'stuff', and use the strongest clamp/vise you can. This will protect the epoxy line as well. Finally, when you do have a hard-to-glue surface, sometimes this can be improved by coating it with a thin layer of a material that will adhere to the G-1, such as chromium. You didn't mention dimpling in your sample prep, but this might also help. In my experience dimpling is less preferential than ion milling, and can help you get a thinner sample that requires less milling.
Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
jds451-at-psu.edu 06/09/2006 09:49 AM Please respond to jds451-at-psu.edu
To Leslie E Krupp/Almaden/IBM-at-IBMUS cc
Subject [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep
---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hi all,
I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the
Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a
press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated.
Thank You,
Jennifer Ray Sloppy Materials Science & Engineering Department Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 9, 17 -- for |--microscopy-at-microscopy.com--|; Fri, 9 Jun 2006 11:45:10 -0500 9, 17 -- Received: (from webmail-at-localhost) 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Message-Id: |--200606091645.MAA01431-at-webmail15.cac.psu.edu--| 9, 17 -- To: microscopy-at-microscopy.com 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 9, 17 -- From: "Jen Sloppy" |--jds451-at-psu.edu--| 9, 17 -- X-Mailer: Penn State WebMail 9, 17 -- X-Sender: jds451 9, 17 -- X-Originating-IP: 128.118.37.54 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
|--br--||--font size=2 face="sans-serif"--|Hi Jennifer-|--/font--| |--br--| |--br--||--font size=2 face="sans-serif"--|There have been several good suggestions already, so I will try to add a few hints that have worked for me. First of all, do make sure you are not milling in the same direction as the epoxy line, as that will preferentially mill the epoxy away. Use beam modulation or the equivalent to restrict the beam to milling perpendicular to the interface. Also, I've noticed the thinner the epoxy line, the better the milling. When gluing your two sample pieces together, make sure the surfaces are absolutely clean, check the epoxy (I always use G-1) for specks of 'stuff', and use the strongest clamp/vise you can. This will protect the epoxy line as well. Finally, when you do have a hard-to-glue surface, sometimes this can be improved by coating it with a thin layer of a material that will adhere to the G-1, such as chromium. You didn't mention dimpling in your sample prep, but this might also help. In my experience dimpling is less preferential than ion milling, and can help you get a thinner sample that requires less milling.|--/font--| |--br--| |--br--||--font size=2 face="sans-serif"--|Leslie|--/font--| |--br--| |--br--||--font size=2 face="sans-serif"--|Leslie Krupp (Thompson)|--br--| IBM Almaden Research|--br--| 650 Harry Road, K19/D1|--br--| San Jose, CA 95120-6099|--br--| (408) 927-3856|--/font--| |--br--| |--br--| |--br--| |--table width=100%--| |--tr valign=top--| |--td width=40%--||--font size=1 face="sans-serif"--||--b--|jds451-at-psu.edu|--/b--| |--/font--| |--p--||--font size=1 face="sans-serif"--|06/09/2006 09:49 AM|--/font--| |--table border--| |--tr valign=top--| |--td bgcolor=white--| |--div align=center--||--font size=1 face="sans-serif"--|Please respond to|--br--| jds451-at-psu.edu|--/font--||--/div--||--/table--| |--br--| |--td width=59%--| |--table width=100%--| |--tr valign=top--| |--td--| |--div align=right--||--font size=1 face="sans-serif"--|To|--/font--||--/div--| |--td--||--font size=1 face="sans-serif"--|Leslie E Krupp/Almaden/IBM-at-IBMUS|--/font--| |--tr valign=top--| |--td--| |--div align=right--||--font size=1 face="sans-serif"--|cc|--/font--||--/div--| |--td--| |--tr valign=top--| |--td--| |--div align=right--||--font size=1 face="sans-serif"--|Subject|--/font--||--/div--| |--td--||--font size=1 face="sans-serif"--|[Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep|--/font--||--/table--| |--br--| |--table--| |--tr valign=top--| |--td--| |--td--||--/table--| |--br--||--/table--| |--br--| |--br--| |--br--||--tt--||--font size=2--||--br--| |--br--| |--br--| ----------------------------------------------------------------------------|--br--| The Microscopy ListServer -- CoSponsor: The Microscopy Society of America|--br--| To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver|--br--| On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html|--br--| ----------------------------------------------------------------------------|--br--| |--br--| Hi all,|--br--| |--br--| I have been attempting to prepare cross sectional TEM samples of anodic tantala|--br--| (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an|--br--| approximate thickness of 200um. The oxide film is grown anodically on the Ta|--br--| using electrochemistry. |--br--| |--br--| In the literature, I have seen cross sectional TEM images of anodic tantala|--br--| grown on sputter deposited tantalum; an aluminum or silicon substrate is|--br--| typically used. I am curious if anyone has had success in preparing anodic|--br--| Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)|--br--| |--br--| My main problem seems to be with de-lamination of the cross section sandwich.|--br--| Here is my attempted procedure: |--br--| |--br--| I first bound two faces of my sample together and placed the sandwich in a press|--br--| while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich|--br--| frequently breaks apart when I begin polishing. To address this problem, I|--br--| encased my sandwich in a brass tube and fitted the tube with supporting brass|--br--| shims, all held in place with epoxy. Using this technique, I was able to thin|--br--| some parts of the specimen to 20-50 um, but the epoxy was no longer present at|--br--| this point. When I attempted to ion-mill, the sandwiches splintered apart, and|--br--| any of the oxide that may have been surviving was obliterated in the ion mill.|--br--| |--br--| Any suggestions or advice would be greatly appreciated.|--br--| |--br--| Thank You,|--br--| |--br--| Jennifer Ray Sloppy|--br--| Materials Science & Engineering Department|--br--| Ph.D. Candidate|--br--| Pennsylvania State University|--br--| |--br--| |--br--| ==============================Original Headers==============================|--br--| 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006|--br--| 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201])|--br--| 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752|--br--| 9, 17 -- for <microscopy-at-microscopy.com>; Fri, 9 Jun 2006 11:45:10 -0500|--br--| 9, 17 -- Received: (from webmail-at-localhost)|--br--| 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431;|--br--| 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT)|--br--| 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT)|--br--| 9, 17 -- Message-Id: <200606091645.MAA01431-at-webmail15.cac.psu.edu>|--br--| 9, 17 -- To: microscopy-at-microscopy.com|--br--| 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep|--br--| 9, 17 -- From: "Jen Sloppy" <jds451-at-psu.edu>|--br--| 9, 17 -- X-Mailer: Penn State WebMail|--br--| 9, 17 -- X-Sender: jds451|--br--| 9, 17 -- X-Originating-IP: 128.118.37.54|--br--| 9, 17 -- MIME-Version: 1.0|--br--| 9, 17 -- Content-Type: text/plain|--br--| ==============================End of - Headers==============================|--br--| |--/font--||--/tt--| |--br--| --=_alternative 005BDC628825718B_=--
==============================Original Headers============================== 33, 44 -- From lkrupp-at-us.ibm.com Mon Jun 12 11:45:33 2006 33, 44 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 33, 44 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CGjWFJ030320 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 11:45:33 -0500 33, 44 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 33, 44 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGjWqG004007 33, 44 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 12:45:32 -0400 33, 44 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 33, 44 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5CGjW0x268382 33, 44 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 12:45:32 -0400 33, 44 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 33, 44 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5CGjVvI008729 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 12:45:31 -0400 33, 44 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 33, 44 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGjVZ8008714; 33, 44 -- Mon, 12 Jun 2006 12:45:31 -0400 33, 44 -- In-Reply-To: |--200606091649.k59GnIlh004856-at-ns.microscopy.com--| 33, 44 -- MIME-Version: 1.0 33, 44 -- X-MIMETrack: S/MIME Sign by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:15 AM, 33, 44 -- Serialize by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:15 AM, 33, 44 -- Serialize complete at 06/12/2006 09:43:15 AM, 33, 44 -- S/MIME Sign failed at 06/12/2006 09:43:15 AM: The cryptographic key was not 33, 44 -- found, 33, 44 -- S/MIME Sign by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:22 AM, 33, 44 -- Serialize by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:22 AM, 33, 44 -- Serialize complete at 06/12/2006 09:43:22 AM, 33, 44 -- S/MIME Sign failed at 06/12/2006 09:43:22 AM: The cryptographic key was not 33, 44 -- found, 33, 44 -- Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 33, 44 -- 06/12/2006 12:45:30, 33, 44 -- Serialize complete at 06/12/2006 12:45:30 33, 44 -- To: jds451-at-psu.edu, Microscopy-at-microscopy.com 33, 44 -- Subject: Re: [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep 33, 44 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 33, 44 -- Message-ID: |--OF2B7D685D.EA6215AC-ON8525718B.005B2D78-8825718B.005BDC65-at-us.ibm.com--| 33, 44 -- From: Leslie E Krupp |--lkrupp-at-us.ibm.com--| 33, 44 -- Date: Mon, 12 Jun 2006 12:45:27 -0400 33, 44 -- Content-Type: multipart/alternative; boundary="=_alternative 005BDC628825718B_=" ==============================End of - Headers==============================
==============================Original Headers============================== 37, 29 -- From lkrupp-at-us.ibm.com Mon Jun 12 11:52:44 2006 37, 29 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 37, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CGqhxa030442 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 11:52:43 -0500 37, 29 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 37, 29 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGqhDZ018610 37, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:43 -0400 37, 29 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 37, 29 -- by d01relay04.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5CGqhL1140474 37, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:43 -0400 37, 29 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 37, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5CGqgWC001568 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:42 -0400 37, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 37, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGqgYV001558 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:42 -0400 37, 29 -- To: microscopy-at-microscopy.com 37, 29 -- Subject: Re: Tantalum / Ta Oxide Cross Section TEM Sample Prep 37, 29 -- MIME-Version: 1.0 37, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 37, 29 -- Message-ID: {OF138984EA.85B0B778-ON8525718B.005C97EB-8825718B.005D1109-at-us.ibm.com} 37, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 37, 29 -- Date: Mon, 12 Jun 2006 09:52:41 -0700 37, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 37, 29 -- 06/12/2006 12:52:42, 37, 29 -- Serialize complete at 06/12/2006 12:52:42 37, 29 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
Many thanks to all the folks who responded to my post. Here is a summary of the suggestions.
At least 4 or 5 people suggested sonicating the powders in a solvent prior to disbursing. After this step, some suggested putting a drop on a filmed grid, or using some type of atomizer to spray the dispersion onto a grid. (Perfume sprayer, air can held in a jar, nebulizer)
A couple people recommended simply mixing the powder with a solvent without sonicating and either letting a drop dry on the grid, or heating it to aid evaporation.
There were two mentions of dry loading the grid, which is dumping some of the powder on the grid and tapping off the excess. This is the method I tried first as it was the easiest, but I have not looked at the grid yet to see if there is enough material to do EELS on.
Thanks again, Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 7, 29 -- From lkrupp-at-us.ibm.com Mon Jun 12 13:11:36 2006 7, 29 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CIBZl6009945 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 13:11:36 -0500 7, 29 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 7, 29 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CIBZMS030830 7, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:11:35 -0400 7, 29 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 7, 29 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5CIA9EN157808 7, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:10:09 -0400 7, 29 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 7, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5CIA9B0006658 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:10:09 -0400 7, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 7, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CIA8Mr006651 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:10:08 -0400 7, 29 -- To: microscopy-at-microscopy.com 7, 29 -- Subject: Powder sample prep summary 7, 29 -- MIME-Version: 1.0 7, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 7, 29 -- Message-ID: {OF7E95A364.DAB1C69A-ON8525718B.005CF6C7-8825718B.00642817-at-us.ibm.com} 7, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 7, 29 -- Date: Mon, 12 Jun 2006 11:10:07 -0700 7, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 7, 29 -- 06/12/2006 14:10:08, 7, 29 -- Serialize complete at 06/12/2006 14:10:08 7, 29 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
Great suggestions, John, I will give them a try. And point well taken regarding the indirect relation of the 2D feature to the 3D pore size. The ASTM method we use (#3576) makes that dimensional correction, but only by assuming close-packed, spherical features. That said, our customers have real use for comparative data from their less ideal samples. If we can select their 2D open cells, they can compare one region to another as a function of molding conditions, one sample to another as a function of processing or R&D variations, or see how they stack up with the competition. But if I can get your suggestions to work, we can offer them stereological analysis as well. Just have to see if they're willing to pay...Thanks again! Yours, Matt
-----Original Message----- X-from: drjohnruss-at-aol.com [mailto:drjohnruss-at-aol.com] Sent: Monday, June 12, 2006 9:10 AM To: microscopy-at-sparc5.microscopy.com Cc: stephenson-at-impactanalytical.com
Greetings listers. We have an image analysis problem, and I could use your help. We are frequently asked to characterize the cell size and structure of various polymer foams for our customers. To date we have relied upon an ASTM method that involves counting the number of cell intersections with a reference line drawn on the image to compute average cell size. The method assumes a degree of homogeneity and cell density that is often an imperfect representation of our customer's samples, and in any event it would be far more desirable to select and measure the cells directly with our ImageProPlus software. There's the rub. We're having the devil of a time getting good segmentation of the open cells against the background matrix. We've tried a number of filters and operations, but thus far without success. I have posted two examples through the link listed below (I disabled it to get through the spam filter). To download, just right click on the file name and "save target as".
**http://www.impactanalytical.com/data/**
If any of you experts out there can set us on the right track, or alternatively explain why we're wasting our time, I would GREATLY appreciate it. My head hurts and the walls are in need of repair!
Incidentally, I have tried varying kV, and collecting in backscatter as well as backscatter plus/minus for topography. Thus far these efforts have not solved the fundamental problem that contrast along the rim of a given cell shifts from light to dark, confounding segmentation. Thanks for your consideration - there's a windy city beer in the bargain if my travel request goes through! Yours, Matt Matthew Stephenson Impact Analytical/MMI 1910 W. Saint Andrews Rd. Midland, MI 48640 (989)832-5555 X506
==============================Original Headers============================== 1, 14 -- From stephenson-at-impactanalytical.com Mon Jun 12 07:19:49 2006 1, 14 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CCJnEA003629 1, 14 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Jun 2006 07:19:49 -0500 1, 14 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 1, 14 -- id {M30M2R2Q} ; Mon, 12 Jun 2006 08:19:47 -0400 1, 14 -- Message-ID: {089C7709BE9235448E3622C6F38D828F06C69196-at-mitconexch.mitcon.org} 1, 14 -- From: "Stephenson, Matthew" {stephenson-at-impactanalytical.com} 1, 14 -- To: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 1, 14 --
Hi All,
I have a user who has presented a very tough problem.
They are looking at a polymer tube (ID ~ .1 - 1 micron) which is filled with a liquid with CNT suspended in it. The tube is the result of drawing from a larger tube and they are hoping to have aligned CNT. They would like to verify this in some way.
I have access to a FESEM and a conventional TEM. The SEM is not environmental. I could not come up with a way to image these. The SEM will not be able to resolve the CNT in any case I think. The TEM could but the sample would have to be thinned to the point that it is likely the inner core of the tube is penetrated. I don't think liquid in the TEM is viable.
I have suggested XRD as a possibility, but the CNT signal on top of the tube and liquid signal may be problematic.
Any thoughts?
Thanks,
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
==============================Original Headers============================== 10, 23 -- From murraytm-at-u.washington.edu Mon Jun 12 14:42:08 2006 10, 23 -- Received: from mxout7.cac.washington.edu (mxout7.cac.washington.edu [140.142.32.178]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CJg7v8000335 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:42:08 -0500 10, 23 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.33.9]) 10, 23 -- by mxout7.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg7wC030917 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:42:07 -0700 10, 23 -- X-Auth-Received: from [128.95.118.89] (tstoebe-mse.ad.engr.washington.edu [128.95.118.89]) 10, 23 -- (authenticated authid=murraytm) 10, 23 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg4aU013369 10, 23 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:42:07 -0700 10, 23 -- Mime-Version: 1.0 (Apple Message framework v749.3) 10, 23 -- X-Priority: 3 10, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 10, 23 -- Message-Id: {427C2B76-E9DC-4F69-9B49-0256CEBCB34D-at-u.washington.edu} 10, 23 -- Content-Transfer-Encoding: 7bit 10, 23 -- From: Tom Murray {murraytm-at-u.washington.edu} 10, 23 -- Subject: Imaging Carbon Nanotubes 10, 23 -- Date: Mon, 12 Jun 2006 12:42:03 -0700 10, 23 -- To: "Microscopy ListServer (E-mail)" {Microscopy-at-microscopy.com} 10, 23 -- X-Mailer: Apple Mail (2.749.3) 10, 23 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_NOT_1 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __HAS_X_PRIORITY 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0' ==============================End of - Headers==============================
Any suggestions on the best way to image protein crystals? They are 20-100 um in length and very stable in high molarity salt solutions or PEG solutions, but dissolve readily in less dilute aqueous solutions. We would like to avoid chemical fixation.
Joe Neilly
==============================Original Headers============================== 2, 18 -- From joe.p.neilly-at-abbott.com Mon Jun 12 15:33:39 2006 2, 18 -- Received: from abtmx3.abbott.com (abtmx3.abbott.com [130.36.44.93]) 2, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CKXdov011623 2, 18 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 15:33:39 -0500 2, 18 -- Received: from abtapn561.northamerica.intra.abbott.com (abtapn561.northamerica.intra.abbott.com [10.236.188.103]) 2, 18 -- by abtmx3.abbott.com (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5CKXcRN008161 2, 18 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 15:33:38 -0500 (CDT) 2, 18 -- To: microscopy-at-microscopy.com 2, 18 -- Subject: Imaging Protein Crystals 2, 18 -- MIME-Version: 1.0 2, 18 -- X-Mailer: Lotus Notes Release 6.5.4 HF761 November 21, 2005 2, 18 -- Message-ID: {OF00F34D2B.F71960B2-ON8625718B.00707FED-8625718B.0070F0E1-at-abbott.com} 2, 18 -- From: Joseph P Neilly {joe.p.neilly-at-abbott.com} 2, 18 -- Date: Mon, 12 Jun 2006 15:33:01 -0500 2, 18 -- X-MIMETrack: Serialize by Router on ABTAPN561/ESVR/ABBOTT(Release 6.5.4FP2|September 12, 2005) at 2, 18 -- 06/12/2006 15:33:33, 2, 18 -- Serialize complete at 06/12/2006 15:33:33 2, 18 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
Greetings, If the goal is not to image the tubes per se but to tell whether they have a net alignment, polarized light microscopy could work. I think sensitivity is ok (With a good instrument, you can detect birefringent retardation of 1 nm and with a really good instrument you can get down to almost 0.1 nm). I expect the problem will come from the polymer tube. Its material could be birefringent and if the inner diameter is really 0.1 micron (rather than 1 micron) then that will probably be too small. But if your i.d.s are closer to 1 micron and the polymer tube itself behaves in the light, it could work...
Good luck! Tobias } } } Hi All, } } I have a user who has presented a very tough problem. } } They are looking at a polymer tube (ID ~ .1 - 1 micron) which is } filled with a liquid with CNT suspended in it. The tube is the } result of drawing from a larger tube and they are hoping to have } aligned CNT. They would like to verify this in some way. } } I have access to a FESEM and a conventional TEM. The SEM is not } environmental. I could not come up with a way to image these. The } SEM will not be able to resolve the CNT in any case I think. The TEM } could but the sample would have to be thinned to the point that it is } likely the inner core of the tube is penetrated. I don't think } liquid in the TEM is viable. } } I have suggested XRD as a possibility, but the CNT signal on top of } the tube and liquid signal may be problematic. } } Any thoughts? } } Thanks, } } 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 } } } ==============================Original Headers============================== } 10, 23 -- From murraytm-at-u.washington.edu Mon Jun 12 14:42:08 2006 } 10, 23 -- Received: from mxout7.cac.washington.edu } (mxout7.cac.washington.edu [140.142.32.178]) } 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k5CJg7v8000335 } 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 } 14:42:08 -0500 } 10, 23 -- Received: from smtp.washington.edu (smtp.washington.edu } [140.142.33.9]) } 10, 23 -- by mxout7.cac.washington.edu } (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg7wC030917 } 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 } 12:42:07 -0700 } 10, 23 -- X-Auth-Received: from [128.95.118.89] } (tstoebe-mse.ad.engr.washington.edu [128.95.118.89]) } 10, 23 -- (authenticated authid=murraytm) } 10, 23 -- by smtp.washington.edu } (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg4aU013369 } 10, 23 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) } 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 } 12:42:07 -0700 } 10, 23 -- Mime-Version: 1.0 (Apple Message framework v749.3) } 10, 23 -- X-Priority: 3 } 10, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 10, 23 -- Message-Id: {427C2B76-E9DC-4F69-9B49-0256CEBCB34D-at-u.washington.edu} } 10, 23 -- Content-Transfer-Encoding: 7bit } 10, 23 -- From: Tom Murray {murraytm-at-u.washington.edu} } 10, 23 -- Subject: Imaging Carbon Nanotubes } 10, 23 -- Date: Mon, 12 Jun 2006 12:42:03 -0700 } 10, 23 -- To: "Microscopy ListServer (E-mail)" {Microscopy-at-microscopy.com} } 10, 23 -- X-Mailer: Apple Mail (2.749.3) } 10, 23 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, } Report='__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_NOT_1 0, __CT 0, } __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, } __HAS_X_PRIORITY 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, } __SANE_MSGID 0, __STOCK_CRUFT 0' } ==============================End of - Headers==============================
On Jun 12, 2006, at 1:33 PM, joe.p.neilly-at-abbott.com wrote:
} Any suggestions on the best way to image protein crystals? They are } 20-100 um in length and very stable in high molarity salt solutions or } PEG } solutions, but dissolve readily in less dilute aqueous solutions. We } would like to avoid chemical fixation. } Dear Joe, CryoTEM is a good method if the crystals are thin enough--10s of nm. They would appear lighter against the darker buffer if the salt concentration is high enough, but they might have very little contrast in a PEG buffer. If the crystals are well-ordered, you should see strong diffraction spots and strong spots in the FFT of an image. Plunge freezing would be good for thin crystals. 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 Mon Jun 12 16:25:51 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 k5CLPoGs032600 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 12 Jun 2006 16:25:51 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 438BB109B07 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 12 Jun 2006 14:25:50 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id E83C33546E 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 12 Jun 2006 14:25:43 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606122033.k5CKXmWv011796-at-ns.microscopy.com} 4, 22 -- References: {200606122033.k5CKXmWv011796-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {7af473644f8ee4dcc48438e217b44feb-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Imaging Protein Crystals 4, 22 -- Date: Mon, 12 Jun 2006 14:37:12 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
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==============================Original Headers============================== 9, 16 -- From zaluzec-at-aaem.amc.anl.gov Mon Jun 12 16:51:05 2006 9, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 9, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CLp3Ho010662 9, 16 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 16:51:04 -0500 9, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 9, 16 -- by aaem.amc.anl.gov (8.12.11.20060308/8.12.10) with ESMTP id k5CLouFN027186 9, 16 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 16:50:57 -0500 9, 16 -- Mime-Version: 1.0 9, 16 -- Message-Id: {p06110404c0b391c03333-at-[146.139.72.105]} 9, 16 -- In-Reply-To: {200606122033.k5CKXdgd011630-at-ns.microscopy.com} 9, 16 -- References: {200606122033.k5CKXdgd011630-at-ns.microscopy.com} 9, 16 -- Date: Mon, 12 Jun 2006 16:50:55 -0500 9, 16 -- To: microscopy-at-microscopy.com 9, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 9, 16 -- Subject: Re: [Microscopy] Imaging Protein Crystals 9, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mhunt-at-flinnsci.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, May 26, 2006 at 09:59:18 ---------------------------------------------------------------------------
Email: mhunt-at-flinnsci.com Name: Maureen Hunt
Organization: Flinn
Education: 9-12th Grade High School
Location: Batavia, Illinois, USA
Question: I am a former materials microscopist and teacher who is trying to find a more exciting way to teach mitosis and meiosis. Currently, everyone uses prepared slides of whitefish blastula to teach animal mitosis and either prepared slides or unprepared slides of allium root tips to teach plant mitosis. Is there a protist or other "animal-like" eukaryote that can be given to the students, squashed and stained to show animal mitosis? I have googled and come up short. Any suggestions? Thank-you for your time.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sullivanmassi-at-comcast.net) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 12, 2006 at 09:12:29 ---------------------------------------------------------------------------
Question: Do you have any recommendations on purchasing microscopes for high school biology? Is there a particular manufacturer that you would go with in terms of value, reliability, etc? Are there new technologies that would fall within a typical high school budget to consider?
Since MItosis and Meiosis are both dynamic processes, I would recommend that you look for movies of the processes.I did a Google search on "mitosis video", and came up with about 300,000 hits. The first few looked quite impressive.
I would also recommend that you teach the function rather than the structure. That is, mitosis functions in normal organismal growth, while meiosis has a significant role only in reproduction.
I can't resist one story. After a lecture that I gave on mitosis in a non-majors Biology course, a student came up to me and said "You have finally cleared up a major mystery for me. I've always wondered how it is that the skin completely covers a little baby, and that it still completely covers the adult that the baby becomes. Now I understand"
Date sent: Mon, 12 Jun 2006 22:04:50 -0500 To: jbs-at-temple.edu X-from: mhunt-at-flinnsci.com Send reply to: mhunt-at-flinnsci.com
This is an extremely broad and unbounded question. How much money are you willing to spend and what analysis methods do you want (LM, DF, DIC, PH, POL, etc.)?
I would recommend querying the usenet group about your question, as it comes up frequently.
sci.techniques.microscopy
Many Web apps have Usenet avenues. There is a large number of informed and experienced folks who deal with LMs on a daily basis. Many are quite helpful for non-professional applications. This is good since the opposite can get out of hand for some folks (i.e., too pricey).
gary g.
At 08:07 PM 6/12/2006, you wrote:
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==============================Original Headers============================== 12, 21 -- From gary-at-gaugler.com Mon Jun 12 23:09:20 2006 12, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D49KoL020603 12, 21 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 23:09:20 -0500 12, 21 -- Received: (qmail 22096 invoked from network); 12 Jun 2006 21:09:19 -0700 12, 21 -- Received: by simscan 1.1.0 ppid: 22093, pid: 22094, t: 0.1669s 12, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 21 -- by qsmtp3 with SMTP; 12 Jun 2006 21:09:19 -0700 12, 21 -- Message-Id: {7.0.1.0.2.20060612210013.024ea218-at-gaugler.com} 12, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 21 -- Date: Mon, 12 Jun 2006 21:09:21 -0700 12, 21 -- To: sullivanmassi-at-comcast.net 12, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 21 -- Subject: Re: [Microscopy] AskAMicroscopist: purchasing microscopes for 12, 21 -- high school biology 12, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 21 -- In-Reply-To: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- References: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- Mime-Version: 1.0 12, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (Goinoutonalamb-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 12, 2006 at 23:04:35 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both Goinoutonalamb-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: Goinoutonalamb-at-yahoo.com Name: Michael Lamb
Organization: Nassau Community College
Education: Undergraduate College
Location: Upton, New York
Title: Tomographic Reconstruction
Question: I am collaborating with a group on a 3D visualization project through Brookhaven National Labs and I have a question pertaining to 3D reconstruction by means of electron tomography. From the literature that I have read, I've gathered that it's recommended that a 200 kv beam or higher is used when working with sections of 120 nm or thicker. Unfortunately, the philips EM-300 that we are working with is only capable of producing a 100 kv beam. Because of the time constraints of our project (10 weeks) it will be very difficult to determine what section thicknesses/maximum tilt angles will work with 100kv with trial and error. Is there a formula, or a guideline of some sort to follow to determine how much mass a 100kv beam can pass with minimal loss of resolution? We are planning on doing a tilt series from -60 to +60 degrees, in 2 degree increments, at 120 nm sections (hopefully). If anyone could lend me some insight as to whether or not this is feasible, and how far we can go, I would greatly appreciate it!
I'm sure there are several articles (also in books, like the one from Joachim Frank on electron tomography) in which the problem of specimen thickness vs. resolution has been discussed.
see e.g. the following article:
Biophys J, February 1998, p. 1031-1042, Vol. 74, No. 2
Electron Tomography of Ice-Embedded Prokaryotic Cells Rudo Grimm,* Hapreet Singh,* Reinhard Rachel,# Dieter Typke,* Wolfram Zillig,* and Wolfgang Baumeister*
best regards, Reinhard Rachel --------------------- PD Dr.Reinhard Rachel Universität Regensburg Zentrum für EM der NWF III am Lehrstuhl fuer Anatomie D-93053 Regensburg - Germany fon +49 941 943 1666, 1720 fax +49 941 943 2868
==============================Original Headers============================== 7, 27 -- From reinhard.rachel-at-biologie.uni-regensburg.de Tue Jun 13 01:49:39 2006 7, 27 -- Received: from rrzmta2.rz.uni-regensburg.de (rrzmta2.rz.uni-regensburg.de [132.199.1.17]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5D6ncdB017577 7, 27 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 01:49:39 -0500 7, 27 -- Received: from rrzmta2.rz.uni-regensburg.de (localhost [127.0.0.1]) 7, 27 -- by localhost (Postfix) with SMTP id 707C3432F3; 7, 27 -- Tue, 13 Jun 2006 08:49:40 +0200 (CEST) 7, 27 -- Received: from listserv.uni-regensburg.de (titan5.rz.uni-regensburg.de [132.199.4.105]) 7, 27 -- by rrzmta2.rz.uni-regensburg.de (Postfix) with ESMTP id 59096434F0; 7, 27 -- Tue, 13 Jun 2006 08:49:40 +0200 (CEST) 7, 27 -- Received: from TITAN5/SpoolDir by listserv.uni-regensburg.de (Mercury 1.48); 7, 27 -- 13 Jun 06 08:49:38 +0200 7, 27 -- Received: from SpoolDir by TITAN5 (Mercury 1.48); 13 Jun 06 08:49:23 +0200 7, 27 -- From: "Reinhard Rachel" {reinhard.rachel-at-biologie.uni-regensburg.de} 7, 27 -- Organization: Universitaet Regensburg 7, 27 -- To: microscopy-at-microscopy.com 7, 27 -- Date: Tue, 13 Jun 2006 08:49:18 +0200 7, 27 -- MIME-Version: 1.0 7, 27 -- Subject: Tomographic Reconstruction: section thickness 7, 27 -- Cc: Goinoutonalamb-at-yahoo.com 7, 27 -- Priority: normal 7, 27 -- X-mailer: Pegasus Mail for Windows (4.31) 7, 27 -- Content-type: text/plain; charset=ISO-8859-1 7, 27 -- Content-description: Mail message body 7, 27 -- Message-Id: {20060613064940.59096434F0-at-rrzmta2.rz.uni-regensburg.de} 7, 27 -- Content-Transfer-Encoding: 8bit 7, 27 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k5D6ncdB017577 ==============================End of - Headers==============================
Probably the most interesting way to teach mitosis is to synchronize cells in culture. It is probably not the easiest though ;-). It is probable that unsynchronized but very actively growing cells will show enough mitotic events to cover the whole cycle. Most of the time they are easy to recognize by normal phase constrast, although fluorescent staining like DAPI are very fast to perform and very impressive to observe. I must admit that after working now for more than 10 years with cells in culture, seeing them split in late telophase is still an exciting event. I witness events which are actually occuring in my own body at that very moment!!!
I agree that you should insist on the functional part though. It is not easy to comprehend these phenomenon in themself, but understand the difference between mitosis and meiosis is really a hard task.
Good luck
Stephane
--- mhunt-at-flinnsci.com wrote:
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==============================Original Headers============================== 10, 20 -- From nizets2-at-yahoo.com Tue Jun 13 03:02:15 2006 10, 20 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D82EIE028952 10, 20 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 03:02:15 -0500 10, 20 -- Received: (qmail 61561 invoked by uid 60001); 13 Jun 2006 08:02:14 -0000 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 20 -- s=s1024; d=yahoo.com; 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 10, 20 -- b=tOIkUDC8s+gOtnIjmKkJvWbDTGAQj+LFZnsv1ozdZzAzDVLXwfVMD/L2Wg73iyegOLD44ZKSqYVGjNScvzxwgYuXu9AWm8tLly+hjFfmeL0wkxd9mT+mpTwPjeXPGCz7M934lDezDdmiDR+9C3R1aYtmYl6GWYsoE0iyVEXRny4= ; 10, 20 -- Message-ID: {20060613080214.61559.qmail-at-web37413.mail.mud.yahoo.com} 10, 20 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Tue, 13 Jun 2006 01:02:14 PDT 10, 20 -- Date: Tue, 13 Jun 2006 01:02:14 -0700 (PDT) 10, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: trying to find a more exciting way to teach 10, 20 -- To: mhunt-at-flinnsci.com 10, 20 -- Cc: microscopy-at-microscopy.com 10, 20 -- In-Reply-To: {200606130311.k5D3B2ck012854-at-ns.microscopy.com} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 10, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
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 iztok.dogsa-at-ijs.si as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: iztok.dogsa-at-ijs.si Name: iztok dogsa
Organization: IJS
Title-Subject: [Filtered] confocal spinning disk microscope
Question: Has anyone experince with confocal spinning disk microscopy?
Which system would you suggest?
We'd like to achieve really high frame rates, but don't want to buy the laser. The arc lamp is Ok for us.
What about BD CARV II imaging system? Is it better than olympus DSU?
We get asked this fairly regularly by schools and parents (so we have stock replies). Often professional microscopists aren't the best source of information for cheap microscope's as all our optical systems are always over Ł10,000, and most lab mainstays like our three confocal microscopes come in at nearer Ł200,000 each (and users still complain about image quality).
Cheap microscopes under Ł100 are always a disappointment and toy (often called student) microscopes can be very poor. You can easily buy second-hand via ebay, but again there are risks that the optics will be damaged or simply very dirty and difficult to clean (look for old branded 'laboratory' microscopes e.g. Bausch & Lomb) and besides you can't really fill a classroom quickly that way, just get a good one for demos. Local labs may have a good old microscope they no longer need and they may give it to schools (or sell them cheaply) if you acknowledge them - do ask parents and write begging letters if you any spare time (but don't expect anything - look for old establishments that will have years of stuff accumulated). Also try any local microscope enthusiast clubs (they may even be persuaded to come in and talk/demo to pupils) and visit other schools for info and a try-out.
There are suppliers geared up to providing microscopes for schools, so ask around at other school's science departments, but expect to pay nearer Ł500 each for a quality setup (although with those like bottom end Meade [www.meade.com] at around Ł100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section. For pond life etc. a stereoscopic 'dissecting' microscope (40x to 120x mag) is ideal, and of course you can get a really long way with a good magnifying glass (not the really small hi-mag cheap lens ones, try before you buy) - I have a few excellent ones at home for Ł1 and a good low mag Osram one that includes an illuminating halogen bulb at Ł8.
Generally prepared slides can rapidly get a bit boring for under 14s, but living or unusual things always attract an audience. Also try your flatbed film scanner (not LiDe and from around Ł60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class can look at a computer screen with some pushing and shoving. Best to try them on 'approval' as many cheap microscopes can be disappointing if you expect too much.
Excellent pre-prepared stained slides of plant stems and leaves or bits of rats, insects etc.. can be bought, but they tend to be expensive and are easily broken by any age-group. Mounted slides keep well though, so 'vintage' ones even from 50 years ago can still look OK - most schools will have some specimen slides knocking about.
At home and our Primary school (under 11) we use the Digital Blue QX-5 (Ł70) - it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. See http://micro.magnet.fsu.edu/optics/intelplay/index.html (not updated for the QX-5 but all applies - the site even discusses ways of contrast enhancement etc..). Once on the PC the 640x480 images can be manipulated and pasted etc, and the QX-5 does time-lapse for things like crystal growth. living plants growing and small animals. The similar but far better built Olympus MIC-D was great but being over Ł500 it was just too expensive for most schools and is now discontinued - there are other similar budget systems about though.
The macro on a good digital camera (like the image stabilised 5MP Canon S2IS going cheap at Ł200 over here - http://www.dpreview.com/reviews/canons2is) is also worth a try, particularly with a small tripod and halogen bendy desk lamp if very close-up, but I'm not sure I'd like a class with 20 boys near my Olympus E500 digital SLR system though. You can get quite reasonable pictures by resting a small compact digital camera lens against the eyepiece of a microscope.
By the way do try growing crystals on a slide, a few drops of a saturated solution of salt (NaCl) or copper sulphate will grow superb crystals on the surface of a slide when viewed under a microscope (but it takes a few hours for the crystals to form and they often look best before the liquids all gone). Just make sure they don't drive the objective tips into the solution. It's not biology but its fun.
Regards
Keith
Try looking in Amazon.com for decent microscope books with lots of pictures (and they have a good customer review system). Plus try web searches for general sites like these (and for more specific topics) :
http://www.101science.com/Microscope.htm http://micro.magnet.fsu.edu/ http://www.microscopy-uk.org.uk/index.html http://www.btinternet.com/~stephen.durr/ http://www.mccroneatlas.com http://www.diatoms.co.uk [for fun images made of diatoms]
---------------------------------------------------------- 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: gary-at-gaugler.com [mailto:gary-at-gaugler.com] Sent: 13 June 2006 05:12 To: keith.morris-at-ucl.ac.uk
This is an extremely broad and unbounded question. How much money are you willing to spend and what analysis methods do you want (LM, DF, DIC, PH, POL, etc.)?
I would recommend querying the usenet group about your question, as it comes up frequently.
sci.techniques.microscopy
Many Web apps have Usenet avenues. There is a large number of informed and experienced folks who deal with LMs on a daily basis. Many are quite helpful for non-professional applications. This is good since the opposite can get out of hand for some folks (i.e., too pricey).
gary g.
At 08:07 PM 6/12/2006, you wrote:
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==============================Original Headers============================== 12, 21 -- From gary-at-gaugler.com Mon Jun 12 23:09:20 2006 12, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D49KoL020603 12, 21 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 23:09:20 -0500 12, 21 -- Received: (qmail 22096 invoked from network); 12 Jun 2006 21:09:19 -0700 12, 21 -- Received: by simscan 1.1.0 ppid: 22093, pid: 22094, t: 0.1669s 12, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 21 -- by qsmtp3 with SMTP; 12 Jun 2006 21:09:19 -0700 12, 21 -- Message-Id: {7.0.1.0.2.20060612210013.024ea218-at-gaugler.com} 12, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 21 -- Date: Mon, 12 Jun 2006 21:09:21 -0700 12, 21 -- To: sullivanmassi-at-comcast.net 12, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 21 -- Subject: Re: [Microscopy] AskAMicroscopist: purchasing microscopes for 12, 21 -- high school biology 12, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 21 -- In-Reply-To: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- References: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- Mime-Version: 1.0 12, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 39, 27 -- From keith.morris-at-ucl.ac.uk Tue Jun 13 08:34:16 2006 39, 27 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 39, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DDYFDs002151 39, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 08:34:16 -0500 39, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 39, 27 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 39, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 39, 27 -- id 1Fq922-000671-Gd; Tue, 13 Jun 2006 14:34:10 +0100 39, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 39, 27 -- To: {sullivanmassi-at-comcast.net} 39, 27 -- Cc: {Microscopy-at-microscopy.com} 39, 27 -- Subject: RE: [Microscopy] Re: AskAMicroscopist: purchasing microscopes for 39, 27 -- Date: Tue, 13 Jun 2006 14:34:09 +0100 39, 27 -- Message-ID: {001b01c68eee$0d1cf500$7b865290-at-keithhigrade} 39, 27 -- MIME-Version: 1.0 39, 27 -- Content-Type: text/plain; 39, 27 -- charset="iso-8859-1" 39, 27 -- X-Mailer: Microsoft Office Outlook 11 39, 27 -- thread-index: AcaOn5P2KNX7omNpRVK9M3jp3oFQuwANfAkw 39, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 39, 27 -- In-Reply-To: {200606130412.k5D4CMXC030741-at-ns.microscopy.com} 39, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 39, 27 -- X-UCL-MailScanner: Found to be clean 39, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 39, 27 -- X-Spam-Status: No 39, 27 -- Content-Transfer-Encoding: 8bit 39, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5DDYFDs002151 ==============================End of - Headers==============================
On topic: The May issue of Microscopy Today contained the following joke:
"Scientifically, maybe body cells -do- replace themselves completely in seven years ⒠but, legally, you’re still married."
Ron Anderson, MT Editor
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 } ---------------------------------------------------------------------------- } } Hi, } } Probably the most interesting way to teach mitosis is } to synchronize cells in culture. It is probably not } the easiest though ;-). It is probable that } unsynchronized but very actively growing cells will } show enough mitotic events to cover the whole cycle. } Most of the time they are easy to recognize by normal } phase constrast, although fluorescent staining like } DAPI are very fast to perform and very impressive to } observe. } I must admit that after working now for more than 10 } years with cells in culture, seeing them split in late } telophase is still an exciting event. I witness events } which are actually occuring in my own body at that } very moment!!! } } I agree that you should insist on the functional part } though. It is not easy to comprehend these phenomenon } in themself, but understand the difference between } mitosis and meiosis is really a hard task. } } Good luck } } Stephane } } --- mhunt-at-flinnsci.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 } } ---------------------------------------------------------------------------- } } } Below is the result of your feedback form } } (NJZFM-ultra-55). It was submitted by } } (mhunt-at-flinnsci.com) from } } } } } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } } } on Friday, May 26, 2006 at 09:59:18 } } } } } --------------------------------------------------------------------------- } } } Email: mhunt-at-flinnsci.com } } Name: Maureen Hunt } } } } Organization: Flinn } } } } Education: 9-12th Grade High School } } } } Location: Batavia, Illinois, USA } } } } Question: I am a former materials microscopist and } } teacher who is trying to find a more exciting way to } } teach mitosis and meiosis. Currently, everyone uses } } prepared slides of whitefish blastula to teach } } animal mitosis and either prepared slides or } } unprepared slides of allium root tips to teach plant } } mitosis. Is there a protist or other "animal-like" } } eukaryote that can be given to the students, } } squashed and stained to show animal mitosis? I have } } googled and come up short. Any suggestions? } } Thank-you for your time. } } } } } } } --------------------------------------------------------------------------- } } } } } ==============================Original } } Headers============================== } } 7, 13 -- From zaluzec-at-ultra5.microscopy.com Mon Jun } } 12 22:04:39 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 } } k5D34caY027718 } } 7, 13 -- for {microscopy-at-microscopy.com} ; Mon, 12 } } Jun 2006 22:04:38 -0500 } } 7, 13 -- Mime-Version: 1.0 } } 7, 13 -- X-Sender: zaluzec-at-ultra5.microscopy.com } } (Unverified) } } 7, 13 -- Message-Id: } } {p06110403c0b3db9f2108-at-[206.69.208.22]} } } 7, 13 -- Date: Mon, 12 Jun 2006 22:04:36 -0500 } } 7, 13 -- To: microscopy-at-microscopy.com } } 7, 13 -- From: mhunt-at-flinnsci.com (by way of } } Ask-A-Microscopist) } } 7, 13 -- Subject: AskAMicroscopist: trying to find } } a more exciting way to teach } } 7, 13 -- mitosis } } 7, 13 -- Content-Type: text/plain; } } charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam } protection around } http://mail.yahoo.com } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 10, 20 -- From nizets2-at-yahoo.com Tue Jun 13 03:02:15 2006 } 10, 20 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) } 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D82EIE028952 } 10, 20 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 03:02:15 -0500 } 10, 20 -- Received: (qmail 61561 invoked by uid 60001); 13 Jun 2006 08:02:14 -0000 } 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 10, 20 -- s=s1024; d=yahoo.com; } 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 10, 20 -- b=tOIkUDC8s+gOtnIjmKkJvWbDTGAQj+LFZnsv1ozdZzAzDVLXwfVMD/L2Wg73iyegOLD44ZKSqYVGjNScvzxwgYuXu9AWm8tLly+hjFfmeL0wkxd9mT+mpTwPjeXPGCz7M934lDezDdmiDR+9C3R1aYtmYl6GWYsoE0iyVEXRny4= ; } 10, 20 -- Message-ID: {20060613080214.61559.qmail-at-web37413.mail.mud.yahoo.com} } 10, 20 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Tue, 13 Jun 2006 01:02:14 PDT } 10, 20 -- Date: Tue, 13 Jun 2006 01:02:14 -0700 (PDT) } 10, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: trying to find a more exciting way to teach } 10, 20 -- To: mhunt-at-flinnsci.com } 10, 20 -- Cc: microscopy-at-microscopy.com } 10, 20 -- In-Reply-To: {200606130311.k5D3B2ck012854-at-ns.microscopy.com} } 10, 20 -- MIME-Version: 1.0 } 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 } 10, 20 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 20 -- From randerson20-at-tampabay.rr.com Tue Jun 13 09:09:14 2006 5, 20 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01.tampabay.rr.com [65.32.5.131]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DE9DBu012872 5, 20 -- for {Microscopy-at-Microscopy.Com} ; Tue, 13 Jun 2006 09:09:13 -0500 5, 20 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 20 -- by ms-smtp-01.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k5DE994f016559; 5, 20 -- Tue, 13 Jun 2006 10:09:12 -0400 (EDT) 5, 20 -- Message-ID: {448EC703.2040702-at-tampabay.rr.com} 5, 20 -- Date: Tue, 13 Jun 2006 10:09:07 -0400 5, 20 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 5, 20 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 5, 20 -- MIME-Version: 1.0 5, 20 -- To: nizets2-at-yahoo.com, Listserver {Microscopy-at-Microscopy.Com} 5, 20 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: trying to find a more exciting 5, 20 -- way to teach 5, 20 -- References: {200606130802.k5D82UNl029179-at-ns.microscopy.com} 5, 20 -- In-Reply-To: {200606130802.k5D82UNl029179-at-ns.microscopy.com} 5, 20 -- Content-Type: text/plain; charset=UTF-8; format=flowed 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
The Olympus with the EM camera is great for high frame rates. No laser. The spinning slits are not as confocal as spinning pinholes, but it is quite bright. I was recently torturing the EM camera and were getting rates that I did not think were possible. Check it out. David
On Jun 13, 2006, at 5:21 AM, iztok.dogsa-at-ijs.si wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://www.microscopy.com/MLFormMail.html } ---------------------------------------------------------------------- } ----- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both iztok.dogsa-at-ijs.si as well as the MIcroscopy } Listserver } ---------------------------------------------------------------------- } ----- } } Email: iztok.dogsa-at-ijs.si } Name: iztok dogsa } } Organization: IJS } } Title-Subject: [Filtered] confocal spinning disk microscope } } Question: Has anyone experince with confocal spinning disk microscopy? } } Which system would you suggest? } } We'd like to achieve really high frame rates, but don't want to buy } the laser. The arc lamp is Ok for us. } } What about BD CARV II imaging system? Is it better than olympus DSU? } } Thanks & regards to you all, } } Iztok } } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Tue Jun 13 07:15:39 2006 } 12, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k5DCFcbR018836 } 12, 12 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 } 07:15:39 -0500 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p0611040ec0b45c3f4728-at-[206.69.208.22]} } 12, 12 -- Date: Tue, 13 Jun 2006 07:15:37 -0500 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: iztok.dogsa-at-ijs.si (by way of MicroscopyListserver) } 12, 12 -- Subject: viaWWW: confocal spinning disk microscope } 12, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
==============================Original Headers============================== 6, 22 -- From Elliott-at-arizona.edu Tue Jun 13 09:58:05 2006 6, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DEw5Rp024151 6, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 09:58:05 -0500 6, 22 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 6, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id A917BE400CD 6, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 07:58:04 -0700 (MST) 6, 22 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 6, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id BBDDBE41125 6, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 07:58:00 -0700 (MST) 6, 22 -- Mime-Version: 1.0 (Apple Message framework v750) 6, 22 -- In-Reply-To: {200606131221.k5DCLHVB025321-at-ns.microscopy.com} 6, 22 -- References: {200606131221.k5DCLHVB025321-at-ns.microscopy.com} 6, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 22 -- Message-Id: {FC930766-D51F-4A66-9CC4-F179FC978836-at-arizona.edu} 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- From: David Elliott {Elliott-at-arizona.edu} 6, 22 -- Subject: Re: [Microscopy] viaWWW: confocal spinning disk microscope 6, 22 -- Date: Tue, 13 Jun 2006 07:57:57 -0700 6, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 6, 22 -- X-Mailer: Apple Mail (2.750) 6, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Thanks to everyone who responded to my question regarding sectioning carbon nanotubes. This listserve is a great resource.
Best wishes,
Soumitra
********************************************************************************** Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282 http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu
==============================Original Headers============================== 5, 20 -- From sghoshro-at-nmsu.edu Tue Jun 13 10:02:51 2006 5, 20 -- Received: from cheech.nmsu.edu (cheech.nmsu.edu [128.123.34.14]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DF2oXd032025 5, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 10:02:51 -0500 5, 20 -- Received: (from nobody-at-localhost) 5, 20 -- by cheech.nmsu.edu (8.11.6/8.11.6) id k5DF2oI04183 5, 20 -- for Microscopy-at-microscopy.com; Tue, 13 Jun 2006 09:02:50 -0600 5, 20 -- Received: from 128.123.105.122 ( [128.123.105.122]) 5, 20 -- as user sghoshro-at-imap.nmsu.edu by webmail.nmsu.edu with HTTP; 5, 20 -- Tue, 13 Jun 2006 09:02:50 -0600 5, 20 -- Message-ID: {1150210970.448ed39a418dd-at-webmail.nmsu.edu} 5, 20 -- Date: Tue, 13 Jun 2006 09:02:50 -0600 5, 20 -- From: Soumitra Ghoshroy {sghoshro-at-nmsu.edu} 5, 20 -- To: Microscopy-at-microscopy.com 5, 20 -- Subject: sectioning carbon nanotubes 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; charset=ISO-8859-1 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- User-Agent: Internet Messaging Program (IMP) 3.0 5, 20 -- X-Originating-IP: 128.123.105.122 ==============================End of - Headers==============================
I'm in the market for a stand alone, oil-less, glow discharge unit. Recommendations would be greatly appreciated.
Thanks, Kim
Kimberly A. Riddle Florida State University Biological Science Imaging Resource 119 Bio Unit I, 4370 Tallahassee, FL 32306 tel: 850.644.6519 fax: 850.644.0481 http://bsir.bio.fsu.edu
==============================Original Headers============================== 8, 21 -- From riddle-at-bio.fsu.edu Tue Jun 13 11:07:07 2006 8, 21 -- Received: from bio.fsu.edu (bio.fsu.edu [128.186.38.55]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DG77pA013652 8, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 11:07:07 -0500 8, 21 -- Received: from riddlepc.bio.fsu.edu (riddlepc.bio.fsu.edu [128.186.14.73]) 8, 21 -- by bio.fsu.edu (8.13.1/8.13.1) with ESMTP id k5DG2ioB085887 8, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 8, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 12:02:44 -0400 (EDT) 8, 21 -- (envelope-from riddle-at-bio.fsu.edu) 8, 21 -- Message-Id: {6.2.5.6.2.20060613120255.02985e88-at-bio.fsu.edu} 8, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 8, 21 -- Date: Tue, 13 Jun 2006 12:06:10 -0400 8, 21 -- To: Microscopy-at-microscopy.com 8, 21 -- From: Kim Riddle {riddle-at-bio.fsu.edu} 8, 21 -- Subject: glow discharge units 8, 21 -- Mime-Version: 1.0 8, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 21 -- X-FSU-Biology-MailScanner: Found to be clean 8, 21 -- X-FSU-Biology-MailScanner-SpamCheck: not spam, SpamAssassin (score=-4.399, 8, 21 -- required 8, autolearn=not spam, ALL_TRUSTED -1.80, BAYES_00 -2.60) 8, 21 -- X-FSU-Biology-MailScanner-From: riddle-at-bio.fsu.edu ==============================End of - Headers==============================
On Jun 13, 2006, at 9:07 AM, riddle-at-bio.fsu.edu wrote:
} I'm in the market for a stand alone, oil-less, glow discharge unit. } Recommendations would be greatly appreciated. } Dear Kim, We have used our Harrick Basic Plasma Cleaner for several trouble-free years, and they also make an Extended Plasma Cleaner, which is a larger model. You would have to buy an oil-free pump to go with either unit, but a diaphragm pump would be adequate for glow-discharging. 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 Jun 13 13:49:31 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 k5DInUSV004811 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 13:49:31 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 71257109A09 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 11:49:30 -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 EE7CC109B2A 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 11:49:24 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606131607.k5DG7FwU013823-at-ns.microscopy.com} 4, 22 -- References: {200606131607.k5DG7FwU013823-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {3476722cb03ade7cd1e8f052a6ab2203-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] glow discharge units 4, 22 -- Date: Tue, 13 Jun 2006 11:49:40 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
I have an individual learning TEM. He will be doing a lot of mouse brain tissue infected with HIV. What sort of safety or handling procedures should we follow? I don't have experience in this area with this sort of pathogen. The tissue is arriving in my lab fixed in 2%glutaraldehyde, 2%paraformaldehyde, 0.5%acrolein in .1M phosphate buffer which I provided to his lab. So, for processing up through embedding can we just handle the samples the same as any other tissue? When sectioning the blocks do the shavings from trimming the block, sectioning, etc. have to be specially collected? If he nicks himself with the razor blade while trimming the block is there an increased risk? Should I have him wearing exam gloves while trimming? Any and all advice will be greatly appreciated.
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center 986395 Nebraska Medical Center Omaha, NE 68198-6395 tbargar-at-unmc.edu 402-559-7347
==============================Original Headers============================== 6, 17 -- From tbargar-at-unmc.edu Tue Jun 13 13:56:53 2006 6, 17 -- Received: from zixvpm01.unmc.edu (zixvpm01.unmc.edu [192.198.54.126]) 6, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DIurm7014950 6, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 13:56:53 -0500 6, 17 -- Received: from zixvpm01.unmc.edu (ZixVPM [127.0.0.1]) 6, 17 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 1395C4C0CE 6, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 13:48:17 -0500 (CDT) 6, 17 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 6, 17 -- by zixvpm01.unmc.edu (Proprietary) with ESMTP id 07BB54C0C9 6, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 13:48:16 -0500 (CDT) 6, 17 -- Subject: TEM of HIV in brain tissue 6, 17 -- To: Microscopy-at-MSA.Microscopy.Com 6, 17 -- Message-ID: {OF5A64AA61.19F4D29B-ON8625718C.0065E182-8625718C.006814A8-at-unmc.edu} 6, 17 -- From: Tom W Bargar {tbargar-at-unmc.edu} 6, 17 -- Date: Tue, 13 Jun 2006 13:56:50 -0500 6, 17 -- MIME-Version: 1.0 6, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Does anyone have a copy of a service manual for the Sorvall MT-2B microtomes? I have the (original) manuals that came with the things, but not a take-it-apart-and-fix-it service manual. If such things still exist, I would appreciate a photocopy. Many thanks!
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 2, 20 -- From oshel1pe-at-cmich.edu Tue Jun 13 15:08:08 2006 2, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DK88C8026809 2, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 15:08:08 -0500 2, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 2, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5DKh34l019524 2, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 16:43:03 -0400 2, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 2, 20 -- Tue, 13 Jun 2006 16:08:07 -0400 2, 20 -- Mime-Version: 1.0 2, 20 -- Message-Id: {f06230913c0b4cb45f81f-at-[141.209.160.132]} 2, 20 -- Date: Tue, 13 Jun 2006 16:08:05 -0400 2, 20 -- To: Microscopy-at-microscopy.com 2, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 2, 20 -- Subject: Sorvall MT-2B service manuals? 2, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 2, 20 -- X-OriginalArrivalTime: 13 Jun 2006 20:08:08.0177 (UTC) FILETIME=[16908610:01C68F25] 2, 20 -- X-CanItPRO-Stream: default 2, 20 -- X-Spam-Score: -4 () L_EXCH_MF 2, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Glutaraldehyde and Formaldehyde are both very good inactivators. When I do level 4 stuff they treat in 4%PF for 48hr to 4 weeks, depending on the protocol they follow at the time, so maybe you want to increase to a more Karnovsky like fix and make the PF 4%. That works for them, it should work for you. Under either condition, by the time you get your stuff it will be well inactivated, and safe.
Paul
==============================Original Headers============================== 4, 21 -- From paul_hazelton-at-umanitoba.ca Tue Jun 13 15:11:29 2006 4, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DKBSB0031349 4, 21 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 15:11:29 -0500 4, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 4, 21 -- (authenticated bits=0) 4, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k5DKBQrf001617; 4, 21 -- Tue, 13 Jun 2006 15:11:26 -0500 (CDT) 4, 21 -- Message-ID: {448F1BEA.8000609-at-umanitoba.ca} 4, 21 -- Date: Tue, 13 Jun 2006 15:11:22 -0500 4, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 4, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 4, 21 -- X-Accept-Language: en-us, en 4, 21 -- MIME-Version: 1.0 4, 21 -- To: tbargar-at-unmc.edu 4, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 4, 21 -- Subject: Re: [Microscopy] TEM of HIV in brain tissue 4, 21 -- References: {200606131858.k5DIwHVW018669-at-ns.microscopy.com} 4, 21 -- In-Reply-To: {200606131858.k5DIwHVW018669-at-ns.microscopy.com} 4, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Question: Hi All, I was wondering if anyone call suggest a marker for endothelial cells in the rat brain (cryostat sections). I wish to count the density of blood vessels in the brains of rat pups who have been exposed to undernutrition or glucocorticoid treatment. I have tried CD31 from Chemicon with little success. I am looking for a structural marker of blood vessels- a marker which will stain all of the blood vessels and not be influenced or altered by undernutrition or hypoxia. Thanks in advance, Emily
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Email: Dandersen-at-optimedica.com Name: Dan Andersen
Organization: OptiMedica
Title-Subject: [Filtered] Histo Prep of ophthalmic samples for EM
Question: I am looking for a private laboraory to process some ocular samples I wish to analyze under EM.
On Jun 12, 2006, at 9:23 PM, Goinoutonalamb-at-yahoo.com wrote:
} Question: I am collaborating with a group on a 3D visualization } project through Brookhaven National Labs and I have a question } pertaining to 3D reconstruction by means of electron tomography. From } the literature that I have read, I've gathered that it's recommended } that a 200 kv beam or higher is used when working with sections of 120 } nm or thicker. Unfortunately, the philips EM-300 that we are working } with is only capable of producing a 100 kv beam. Because of the time } constraints of our project (10 weeks) it will be very difficult to } determine what section thicknesses/maximum tilt angles will work with } 100kv with trial and error. Is there a formula, or a guideline of } some sort to follow to determine how much mass a 100kv beam can pass } with minimal loss of resolution? We are planning on doing a tilt } series from -60 to +60 degrees, in 2 degree increments, at 120 nm } sections (hopefully). If anyone could lend me some insight as to } whether or not this is feasible, and how ! } far we can go, I would greatly appreciate it! } Dear Michael, Some issues not answered by the other person who responded to your post are what the specimen consists of, and what resolution you want. For biological materials and polymers consisting of low-Z materials, the penetration of the beam will be greater than for electronics components consisting of silicon or germanium, and minerals containing high-Z atoms will be more restricted still. On the other hand, some of the restriction on thickness comes from the fact that at high tilt, the effective thickness is greater than the specimen thickness--you mention 60 degrees, at which angle it is twice the thickness. You may be able to get significant information by taking stereo pairs; these are obtained at small tilts, usually {10 degrees, so a thicker specimen can be observed in 3-D through a stereo viewer, which may be all you need, and 3-D reconstructions can be made with such software as sterecon, which was used at the Wadsworth Center in Albany NY when I was there. If your specimen exceeds 120 nm by a small amount, you can still try tomography with an angular range less that +/-60 degrees and obtain useful data (although artifacts from the missing wedge will be more pronounced the smaller the angular range), and you can take dual-axis tilt series, which will reduce the missing wedge to a missing pyramid and lessen the missing data artifacts. Finally, if the Wadsworth Center is still a Regional Resource, you might be able to get time on one of their higher-voltage EMs, and they have a 1.2 MV scope, which should provide sufficient penetrating power even for roughly 1 um sections and/or for higher-Z materials. Good luck. 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 Jun 13 19:38:17 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 k5E0cGUs007753 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 19:38:17 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id A8AF235624 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 17:38:15 -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 C25E7109C6C 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 17:38:09 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606130423.k5D4Nlk9004749-at-ns.microscopy.com} 4, 22 -- References: {200606130423.k5D4Nlk9004749-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {fee7f3cf2eefaec0e4d6d3b0b3620142-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] AskAMicroscopist: Tomographic Reconstruction 4, 22 -- Date: Tue, 13 Jun 2006 17:38:26 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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Email: aribaudo-at-triprinceton.org Name: Anthony Ribaudo
Organization: TRI Princeton
Title-Subject: [Filtered] TEM Sample Preparation of Paper using Foraldehyde
Question: Dear All,
Can anyone suggest the experimental setup, i.e. the apparatus needed to prepare paper samples that utilizes formaldehyde to crosslink the paper fibers. Formaldehyde imparts water-resistant qualities and increases the dimensional stability against moisture. The formaldehyde reaction conditions recommended are as follow: Temperature 140˘C. Reaction period 0.5-48 hours. Formaldehyde concentration introduced in vapor phase 2.19 x 10-2 mol/L.
After treatment the paper sample is embedded in epoxy resin and ultrasectioning.
Thank you,
Anthony Ribaudo TRI Princeton 601 Prospect Avenue Princeton, NJ 08542
I was asked to look into the morphology and cause of tin whiskers in high tin content lead free solders by a guy I meet. Some of the following questions have been asked for 50 years. Maybe someone out there can share some more recent expertise about tin whiskers.
* What is the root cause of crystalline tin whiskers? (It's probably not resolved yet. There are lots of conflicting opinions and observations on Google.)
* How do you prevent tin whiskers from growing out of almost pure tin solder joints? (He is not that interested in a thin film coating solution.) * What is the mechanism(s) that can cause 2 micron diameter or less "fibers" (et al) to grow right out of the surface of solder joins to a length of up to one observed to be 10 mm long? * What are the EDS results of tin whisker analysis? * Are the EDS analysis results settled science? * Have solder joints been ion milled to explore the interior of the solder or is this strictly a compression/stress surface phenomena? Or both?
He is NOT interested in high humidity dendritic growth of tin on PCB surfaces reputed to be caused by electric lines and associated equipotential lines. He is interested in the shorts caused by tin whiskers bridging solder joints through the air or a vacuum.
This raises the following big question. * How do you accelerate the growth of tin whiskers so that they can be studied without waiting 1-15 years for them to develop?
Please reply off list with any insights you have on this problem. I don't think this subject will have wide appeal. Thank you in advance for any perspectives that can be shared with him. If you are speculating on a solution or theory, please say so in your reply.
Paul
Disclaimers: I do not work for this guy's company, the guy, or any company or organization working on tin whiskers. I was astonished at the phenomenon, morphology, and how they ruined an obiting satellite. Tin whiskers grow in air or vacuum.
==============================Original Headers============================== 13, 24 -- From beaurega-at-westol.com Thu Jun 15 09:54:50 2006 13, 24 -- Received: from smtp-gateway-5.winbeam.com (smtp-gateway-5.winbeam.com [64.84.97.70]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FEsnwQ003359 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 09:54:50 -0500 13, 24 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 13, 24 -- by smtp-gateway-5.winbeam.com (8.13.2/8.12.8) with SMTP id k5FEs1LD015079 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 10:54:21 -0400 13, 24 -- Received: (qmail 30540 invoked by uid 89); 15 Jun 2006 14:53:29 -0000 13, 24 -- Received: from pitts-69-72-12-3.dynamic-dialup.coretel.net (HELO millenium) (69.72.12.3) 13, 24 -- by mail.winbeam.com with SMTP; 15 Jun 2006 14:53:29 -0000 13, 24 -- Message-Id: {3.0.6.32.20060615105431.007edaf0-at-pop3.norton.antivirus} 13, 24 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 13, 24 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 13, 24 -- Date: Thu, 15 Jun 2006 10:54:31 -0400 13, 24 -- To: microscopy-at-microscopy.com 13, 24 -- From: Beaurega {beaurega-at-westol.com} 13, 24 -- Subject: Tin Whiskers in high tin content solders. 13, 24 -- Mime-Version: 1.0 13, 24 -- Content-Type: text/plain; charset="us-ascii" 13, 24 -- X-Winbeam-MailScanner-Information: - Please contact Technical Support for more information 13, 24 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) 13, 24 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, score=0, 13, 24 -- required 6, autolearn=disabled) 13, 24 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
You're right. There is not a clear concensus on the formation of tin whiskers. A couple of great sites on the subject are http://www.inemi.org/cms/newsroom/Presentations/TinWhiskerWorkshop_ECTC0 6.html and http://nepp.nasa.gov/whisker/
If you wander around on these sites, you'll find enough reports to read to keep you busy for quite a while. I notice that an April 2006 report from NASA ( http://nepp.nasa.gov/whisker/reference/tech_papers/2006-Leidecker-Tin-Wh isker-Failures.pdf ) states that "If you can't live with tin whiskers, then "Don't Use Tin"."
There are things that will keep whiskers from growing, most notably adding lead (but that probably doesn't help much).
Scratching the surface of the tin will influence more whisker growth (but I'm not sure if it'll be faster whisker growth). There are some extensive studies that have been done to try to find out what accelerates whisker growth, so I'd read those before starting my own study.
Hope this helps.
Diane _____________________________ Diane Ciaburri Principal Materials Engineer General Dynamics 100 Plastics Ave., Rm 2552A Pittsfield MA 01201 phone: (413) 494-3430 email: diane.ciaburri-at-gd-ais.com
-----Original Message----- X-from: beaurega-at-westol.com [mailto:beaurega-at-westol.com] Sent: Thursday, June 15, 2006 10:55 AM To: Ciaburri, Diane A.
Hello,
Anyone with contact info for a 3rd party service person/organization that can provide support services for a Rigaku Model: Ultra-18 XRD instrument, please contact Mr. Ross Potoff at the University of Arizona (520) 621-8101 or email to Potoff-at-email.arizona.edu.
Regards,
Bob Roberts EM Lab Services, Inc. 449 NW 62nd St. Topeka, Kansas 66617-1780 785.246.1232 voice 785.246.0168 fax www.emlabservices.com
==============================Original Headers============================== 6, 23 -- From emlabservices-at-cox.net Thu Jun 15 12:25:28 2006 6, 23 -- Received: from centrmmtao04.cox.net (centrmmtao04.cox.net [70.168.83.80]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FHPQ41028447 6, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 12:25:27 -0500 6, 23 -- Received: from EMLabServices ([24.255.213.54]) by centrmmtao04.cox.net 6, 23 -- (InterMail vM.6.01.06.01 201-2131-130-101-20060113) with SMTP 6, 23 -- id {20060615172523.FNKV14774.centrmmtao04.cox.net-at-EMLabServices} 6, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 13:25:23 -0400 6, 23 -- Message-ID: {006b01c690a0$b0915740$6400a8c0-at-EMLabServices} 6, 23 -- From: "EM Lab Services" {emlabservices-at-cox.net} 6, 23 -- To: {Microscopy-at-microscopy.com} 6, 23 -- Subject: XRD: Service/support for Rigaku 6, 23 -- Date: Thu, 15 Jun 2006 11:25:25 -0600 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- format=flowed; 6, 23 -- charset="iso-8859-1"; 6, 23 -- reply-type=original 6, 23 -- Content-Transfer-Encoding: 7bit 6, 23 -- X-Priority: 3 6, 23 -- X-MSMail-Priority: Normal 6, 23 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 6, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
I would be curious to see any feedback on this subject, in the name of education.
I have a couple questions. Some of the lead-free solders have silver in them. Are you (or he) sure the whiskers are tin, or could they be silver? When you say "vacuum", are you referring to the low pressures in orbit, or a clean, high vacuum in a chamber of some sort? (or, perhaps, vacuum tube electronic component)
I believe the fields associated with a bias have something to do with it, but don't have any facts, or know what the mechanism is. I wonder if a high (strong) bias, in a vacuum chamber, would accelerate the phenomenon.
Looking forward to learning more. Thanks, Darrell
beaurega-at-westol.com wrote on 06/15/2006 10:55:59 AM:
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} } Hi, } } I was asked to look into the morphology and cause of tin whiskers in high } tin content lead free solders by a guy I meet. Some of the following } questions have been asked for 50 years. Maybe someone out there can share } some more recent expertise about tin whiskers. } } * What is the root cause of crystalline tin whiskers? (It's probably not } resolved yet. There are lots of conflicting opinions and observations on } Google.) } } * How do you prevent tin whiskers from growing out of almost pure tin } solder joints? } (He is not that interested in a thin film coating solution.) } * What is the mechanism(s) that can cause 2 micron diameter or less } "fibers" (et al) to grow right out of the surface of solder joins to a } length of up to one observed to be 10 mm long? } * What are the EDS results of tin whisker analysis? } * Are the EDS analysis results settled science? } * Have solder joints been ion milled to explore the interior of the solder } or is this strictly a compression/stress surface phenomena? Or both? } } He is NOT interested in high humidity dendritic growth of tin on PCB } surfaces reputed to be caused by electric lines and associated } equipotential lines. He is interested in the shorts caused by tin whiskers } bridging solder joints through the air or a vacuum. } } This raises the following big question. } * How do you accelerate the growth of tin whiskers so that they can be } studied without waiting 1-15 years for them to develop? } } Please reply off list with any insights you have on this problem. I don't } think this subject will have wide appeal. Thank you in advance for any } perspectives that can be shared with him. } If you are speculating on a solution or theory, please say so in your reply. } } Paul } } Disclaimers: I do not work for this guy's company, the guy, or any company } or organization working on tin whiskers. } I was astonished at the phenomenon, morphology, and how they ruined an } obiting satellite. } Tin whiskers grow in air or vacuum. } } } } } } } ==============================Original Headers============================== } 13, 24 -- From beaurega-at-westol.com Thu Jun 15 09:54:50 2006 } 13, 24 -- Received: from smtp-gateway-5.winbeam.com (smtp-gateway-5. } winbeam.com [64.84.97.70]) } 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k5FEsnwQ003359 } 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 09:54:50 -0500 } 13, 24 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) } 13, 24 -- by smtp-gateway-5.winbeam.com (8.13.2/8.12.8) with SMTP } id k5FEs1LD015079 } 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 10:54:21 -0400 } 13, 24 -- Received: (qmail 30540 invoked by uid 89); 15 Jun 2006 14: } 53:29 -0000 } 13, 24 -- Received: from pitts-69-72-12-3.dynamic-dialup.coretel.net } (HELO millenium) (69.72.12.3) } 13, 24 -- by mail.winbeam.com with SMTP; 15 Jun 2006 14:53:29 -0000 } 13, 24 -- Message-Id: {3.0.6.32.20060615105431.007 edaf0-at-pop3.norton.antivirus} } 13, 24 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus } 13, 24 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } 13, 24 -- Date: Thu, 15 Jun 2006 10:54:31 -0400 } 13, 24 -- To: microscopy-at-microscopy.com } 13, 24 -- From: Beaurega {beaurega-at-westol.com} } 13, 24 -- Subject: Tin Whiskers in high tin content solders. } 13, 24 -- Mime-Version: 1.0 } 13, 24 -- Content-Type: text/plain; charset="us-ascii" } 13, 24 -- X-Winbeam-MailScanner-Information: - Please contact } Technical Support for more information } 13, 24 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) } 13, 24 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin } (notcached, score=0, } 13, 24 -- required 6, autolearn=disabled) } 13, 24 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com } ==============================End of - Headers==============================
==============================Original Headers============================== 10, 29 -- From milesd-at-us.ibm.com Thu Jun 15 15:33:05 2006 10, 29 -- Received: from e3.ny.us.ibm.com (e3.ny.us.ibm.com [32.97.182.143]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FKX5Xm010224 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 15:33:05 -0500 10, 29 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 10, 29 -- by e3.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5FKX3tL022193 10, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:03 -0400 10, 29 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 10, 29 -- by d01relay04.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5FKX1ld264774 10, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:03 -0400 10, 29 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 10, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5FKX1nS002256 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:01 -0400 10, 29 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 10, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5FKX1Ll002235 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:01 -0400 10, 29 -- In-Reply-To: {200606151455.k5FEtxZZ004881-at-ns.microscopy.com} 10, 29 -- Subject: Re: [Microscopy] Tin Whiskers in high tin content solders. 10, 29 -- To: Microscopy-at-MSA.Microscopy.Com 10, 29 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 10, 29 -- Message-ID: {OF7455FAA7.1B918C5E-ON8525718E.006F3FB0-8525718E.0070E1DC-at-us.ibm.com} 10, 29 -- From: Darrell Miles {milesd-at-us.ibm.com} 10, 29 -- Date: Thu, 15 Jun 2006 16:32:59 -0400 10, 29 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 7.0.1 HF4|May 16, 2006) at 10, 29 -- 06/15/2006 16:33:00 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
The whiskers are tin. Silver is added to lead solder to reduce the melting point.
Whiskering occurs at room temperature, high and low humidity and all sorts of conditions. It is a nasty problem for commercial and especially MIL applications. Alternate solders are being studied (different combinations of elements) but their melting point is sufficiently higher than Pb solder to be a real problem.
gary g.
At 01:35 PM 6/15/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Thu Jun 15 16:10:07 2006 10, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5FLA6D2021040 10, 20 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 16:10:06 -0500 10, 20 -- Received: (qmail 5097 invoked from network); 15 Jun 2006 14:10:06 -0700 10, 20 -- Received: by simscan 1.1.0 ppid: 5093, pid: 5094, t: 0.2385s 10, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 20 -- by qsmtp3 with SMTP; 15 Jun 2006 14:10:05 -0700 10, 20 -- Message-Id: {7.0.1.0.2.20060615140636.024fa7d0-at-gaugler.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 20 -- Date: Thu, 15 Jun 2006 14:10:06 -0700 10, 20 -- To: milesd-at-us.ibm.com 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] Re: Tin Whiskers in high tin content solders. 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200606152035.k5FKZcD3013387-at-ns.microscopy.com} 10, 20 -- References: {200606152035.k5FKZcD3013387-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I am in the middle of efforts to transfer manufacturing operations from Lead:Tin solder to Lead Free solder. Presently Silver and copper are used with the tin to reduce the melting point to about 220C. There are many different theories about why tin whiskers form including a compressive stress on the tin layer. The stress may be caused by copper diffusing into the tin at grain boundaries and forming copper:tin complexes which take up more volume.
There is a historical paper covering the history of the research entitled: A History of Tin Whisker Theory: 1946 to 2004 George T. Galyon IBM eSG Group Poughkeepsie, New York
I'm sorry I can't find the original source for the article. It isn't mentioned on my document. But George T. Gaylon has been (and may still be) associated with iNEMI. Some of his material is available through them at http://www.inemi.org/cms/newsroom/NEMI_NIST_TMS_workshop.html
Hope this helps. The electronics industry has done a lot of work in this area in the past five or so years.
Mark Woolley PTRL Lab Avaya, Inc. 1300 West 120th Ave. Westminster, CO 80234 (303) 538-9179 Office (303) 538-2166 Lab woolleym-at-avaya.com
==============================Original Headers============================== 8, 24 -- From woolleym-at-avaya.com Thu Jun 15 18:21:15 2006 8, 24 -- Received: from co300216-ier2.net.avaya.com (co300216-ier2.net.avaya.com [198.152.13.103]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FNLFaH006392 8, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 18:21:15 -0500 8, 24 -- Received: from cof110avexu2.global.avaya.com (h135-9-6-17.avaya.com [135.9.6.17]) 8, 24 -- by co300216-ier2.net.avaya.com (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5FNIbcK023470 8, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 19:18:38 -0400 8, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 24 -- content-class: urn:content-classes:message 8, 24 -- MIME-Version: 1.0 8, 24 -- Content-Type: text/plain; 8, 24 -- charset="iso-8859-1" 8, 24 -- Subject: Tin Whiskers 8, 24 -- Date: Thu, 15 Jun 2006 17:21:14 -0600 8, 24 -- Message-ID: {A49F94D2EEDFB74E816FF4277B7A602C01BF13A2-at-cof110avexu2.global.avaya.com} 8, 24 -- X-MS-Has-Attach: 8, 24 -- X-MS-TNEF-Correlator: 8, 24 -- Thread-Topic: Tin Whiskers 8, 24 -- Thread-Index: AcaQ0mWIOWa4wn+1TAy0Ej7r8a8LEA== 8, 24 -- From: "Woolley, Mark D \(Mark\)" {woolleym-at-avaya.com} 8, 24 -- To: {Microscopy-at-microscopy.com} 8, 24 -- X-Scanner: InterScan AntiVirus for Sendmail 8, 24 -- Content-Transfer-Encoding: 8bit 8, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5FNLFaH006392 ==============================End of - Headers==============================
Hi all. I would like to take a look a the moth eye structure or a fly, but I'm completely new to biological sample... What kind of sample preparation is recommended (any essiccation procedure)? Or it is simple possible to coat it with a metal layer and this will avoid it to explode and splatter around the column? what will happen in the latter case? (just will have to wait longer for a good vacuum next time, or ppermanent damage to any parts?)
Please let me know. Any help is welcome. All the best, Marco.
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr. Marco Salerno, Ph.D.: marco.salerno-at-unile.it CNR-INFM Center NNL: www.nnl.it c/o Palazzine Garrisi, via per Arnesano 16 (km 5) 73100 Lecce - Italy Off. +39-0832-29.8236 Lab.+39-0832-29.8336/8347 Mob.+39-349-26.75.277 Fax +39-0832-29.8238 Home +39-0832-32.51.45 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
==============================Original Headers============================== 6, 22 -- From marco.salerno-at-unile.it Fri Jun 16 02:03:27 2006 6, 22 -- Received: from mailing.unile.it (mailing.unile.it [193.204.77.251]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5G73QZp026611 6, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 02:03:27 -0500 6, 22 -- Received: from MSALERNO (unknown [193.204.74.205]) 6, 22 -- by mailing.unile.it (Postfix) with SMTP id 04BE8104B71 6, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 09:03:25 +0200 (CEST) 6, 22 -- Message-ID: {029d01c69113$067b2e20$cc1e0a0a-at-Utente} 6, 22 -- From: "Marco Salerno" {marco.salerno-at-unile.it} 6, 22 -- To: {Microscopy-at-microscopy.com} 6, 22 -- Subject: first time of a soft matter specimen, please help! 6, 22 -- Date: Fri, 16 Jun 2006 09:03:52 +0200 6, 22 -- MIME-Version: 1.0 6, 22 -- Content-Type: text/plain; 6, 22 -- format=flowed; 6, 22 -- charset="iso-8859-1"; 6, 22 -- reply-type=original 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- X-Priority: 3 6, 22 -- X-MSMail-Priority: Normal 6, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 6, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
you will find some EM analytical labs at this site: http://microanalysis.mikroanalytik.de/service%20microanalysis%20e.phtml
Regards
Frank
Dandersen-at-optimedica.com wrote:
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==============================Original Headers============================== 7, 21 -- From eggert-at-mikroanalytik.de Fri Jun 16 04:48:19 2006 7, 21 -- Received: from Mail1.KONTENT.De (mail1.kontent.de [81.88.34.36]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5G9mIxG008591 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 04:48:18 -0500 7, 21 -- Received: from mikroanalytik.de (p54BDF1C1.dip.t-dialin.net [84.189.241.193]) 7, 21 -- by Mail1.KONTENT.De (Postfix) with ESMTP id 858DE104A3F2; 7, 21 -- Fri, 16 Jun 2006 11:48:19 +0200 (CEST) 7, 21 -- Message-ID: {44927F30.4060702-at-mikroanalytik.de} 7, 21 -- Date: Fri, 16 Jun 2006 11:51:44 +0200 7, 21 -- From: Frank Eggert {eggert-at-mikroanalytik.de} 7, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; de-DE; rv:1.4) Gecko/20030619 Netscape/7.1 (ax) 7, 21 -- X-Accept-Language: de, de-de, en 7, 21 -- MIME-Version: 1.0 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- Cc: Dandersen-at-optimedica.com 7, 21 -- Subject: Re: [Microscopy] viaWWW: private laboraory to process some ocular 7, 21 -- samples 7, 21 -- References: {200606140030.k5E0Um81027205-at-ns.microscopy.com} 7, 21 -- In-Reply-To: {200606140030.k5E0Um81027205-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==============================
I routinely image insects in the SEM without any preparation other than sputter coating. I kill the insects first with ethyl acetate fumes or CO2. Then mount using a double-sticky carbon conductive tab, and coat. Note: if the insect/etc. only contacts the mounting tab in a small area because of convexity, you may have to also use some silver paint to improve the electrical ground. This is commonly needed (something I do automatically). The paint requires a wait to dry. Nor is it always needed -- the University president brought his 6 year-old son to our facility with a beetle to see the SEM. It was a quick mount, no paint, 3 minute Au coat, and in the SEM. Good points for the facility, too. If the critters aren't particularly hairy or you have access to an ESEM, you might be able to leave off the coating. Some insects and arachnids can be imaged at low kVs without coating, and will survive the experience. Note though, that soft-bodied arthropods can (and will) shrink and distort in the vacuum. Robust ones hold up better. Beetles are particularly good for this. Make certain your insect is *intact*. Any missing limbs, antennae, etc. will allow for haemolymph to be sucked out into the chamber. This will cause the body to collapse and coat the innards of your 'scope with bug goo. Not good. I wouldn't do this at all in an instrument used for high-resolution or analytical work. Personally, though, external eye structure is a bit boring. Feet and especially mouthparts are much more interesting. Plus, their morphology is directly related to ecology. Eyes are, yes, but less obviously in external SEM-level anatomy. There are lots of examples on the web, as well as books of example images, with technique chapters.
Phil
} Hi all. } I would like to take a look } a the moth eye structure or a fly, } but I'm completely new to } biological sample... } What kind of sample preparation is recommended } (any essiccation procedure)? } Or it is simple possible to coat it with a } metal layer and this will avoid it to explode and splatter } around the column? } what will happen in the latter case? } (just will have to wait longer for a good vacuum } next time, or ppermanent damage to any parts?) } } Please let me know. } Any help is welcome. } All the best, } Marco. } } } xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx } Dr. Marco Salerno, Ph.D.: } marco.salerno-at-unile.it } CNR-INFM Center NNL: } www.nnl.it } c/o Palazzine Garrisi, } via per Arnesano 16 (km 5) } 73100 Lecce - Italy } Off. +39-0832-29.8236 } Lab.+39-0832-29.8336/8347 } Mob.+39-349-26.75.277 } Fax +39-0832-29.8238 } Home +39-0832-32.51.45 } xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 24 -- From oshel1pe-at-cmich.edu Fri Jun 16 07:40:12 2006 4, 24 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GCeCvW022096 4, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 07:40:12 -0500 4, 24 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 24 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5GDEx4l012931; 4, 24 -- Fri, 16 Jun 2006 09:14:59 -0400 4, 24 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 24 -- Fri, 16 Jun 2006 08:40:09 -0400 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {f06230903c0b851ca1e3b-at-[141.209.160.132]} 4, 24 -- In-Reply-To: {200606160710.k5G7AHYu001950-at-ns.microscopy.com} 4, 24 -- References: {200606160710.k5G7AHYu001950-at-ns.microscopy.com} 4, 24 -- Date: Fri, 16 Jun 2006 08:40:06 -0400 4, 24 -- To: marco.salerno-at-unile.it 4, 24 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 24 -- Subject: Re: [Microscopy] first time of a soft matter specimen, please 4, 24 -- help! 4, 24 -- Cc: Microscopy-at-microscopy.com 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-OriginalArrivalTime: 16 Jun 2006 12:40:10.0015 (UTC) FILETIME=[012BBAF0:01C69142] 4, 24 -- X-CanItPRO-Stream: default 4, 24 -- X-Spam-Score: -4 () L_EXCH_MF 4, 24 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
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Email: mbisher-at-princeton.edu Name: Margaret Bisher
Organization: Princeton University
Title-Subject: [Filtered] TEM of Soil Microbes
Question: Over the summer we offer a short course for high school science teachers in our department (Molecular Biology). Something the instructors would like to incorporate this year, would be to look at, if possible, their samples in the TEM. We did try it last year and met with limited sucess so I thought I would post this question to the group before this year's class. I am wondering if there is a method for them to clean up their "dirt" to get a better yield (I did find some phage and bacteria but I had to spend hours looking, it would have more exciting for them if there been more to look at) and then I can possibly make a suspension and then do negative stain on the sample, to see if we have any bacteria, etc. in the sample.
I'd appreciate any suggestions you might be able to supply.
Thanks, Peggy Bisher
Electron Microscopy & Histology Core Facility Manager Princeton University Department of Molecular Biology Moffett Laboratory, Room 113B Washington Road Princeton, New Jersey 08544 Voice: (609) 258-7026 Fax: (609) 258-8468 Email: mbisher-at-princeton.edu
If this phenomenon can be reliably reproduced, the effect from bias should be easily confirmed or denied.
This appears to be some type of transport mechanism... vapor transport, surface transport, bulk diffusion from the body of the metal, or grain boundary diffusion. It would be interesting to find where tin is being depleted in the solder joint, perhaps using EDS methods and metallographic cross sections. The challenge would be to distinguish tin depletion from normal solder casting segregation.
At the moment I'm assuming the whisker is a single crystal. Perhaps the whisker can be studied using x-ray crystallographic methods - either Laue or TEM methods. This may give a clue to the growth mechanism.
Stu Smalinskas, P.E. Metallurgist SKF Plymouth, Michigan
--- gary-at-gaugler.com wrote:
The whiskers are tin. Silver is added to lead solder to reduce the melting point.
Whiskering occurs at room temperature, high and low humidity and all sorts of conditions. It is a nasty problem for commercial and especially MIL applications. Alternate solders are being studied (different combinations of elements) but their melting point is sufficiently higher than Pb solder to be a real problem.
gary g.
---------------------------------------
At 01:35 PM 6/15/2006, you wrote:
Hi,
I would be curious to see any feedback on this subject, in the name of education.
I have a couple questions. Some of the lead-free solders have silver in them. Are you (or he) sure the whiskers are tin, or could they be silver? When you say "vacuum", are you referring to the low pressures in orbit, or a clean, high vacuum in a chamber of some sort? (or, perhaps, vacuum tube electronic component)
I believe the fields associated with a bias have something to do with it, but don't have any facts, or know what the mechanism is. I wonder if a high (strong) bias, in a vacuum chamber, would accelerate the phenomenon.
Looking forward to learning more. Thanks, Darrell
-----------------------------------------------
beaurega-at-westol.com wrote on 06/15/2006 10:55:59 AM:
Hi, I was asked to look into the morphology and cause of tin whiskers in high tin content lead free solders by a guy I meet. Some of the following questions have been asked for 50 years. Maybe someone out there can share some more recent expertise about tin whiskers.
* What is the root cause of crystalline tin whiskers? (It's probably not resolved yet. There are lots of conflicting opinions and observations on Google.)
* How do you prevent tin whiskers from growing out of almost pure tin solder joints? (He is not that interested in a thin film coating solution.)
* What is the mechanism(s) that can cause 2 micron diameter or less "fibers" (et al) to grow right out of the surface of solder joins to a length of up to one observed to be 10 mm long?
* What are the EDS results of tin whisker analysis?
* Are the EDS analysis results settled science?
* Have solder joints been ion milled to explore the interior of the solder or is this strictly a compression/stress surface phenomena? Or both?
He is NOT interested in high humidity dendritic growth of tin on PCB surfaces reputed to be caused by electric lines and associated equipotential lines. He is interested in the shorts caused by tin whiskers bridging solder joints through the air or a vacuum.
This raises the following big question. * How do you accelerate the growth of tin whiskers so that they can be studied without waiting 1-15 years for them to develop?
Please reply off list with any insights you have on this problem. I don't think this subject will have wide appeal. Thank you in advance for any perspectives that can be shared with him. If you are speculating on a solution or theory, please say so in your reply.
Paul
Disclaimers: I do not work for this guy's company, the guy, or any company or organization working on tin whiskers. I was astonished at the phenomenon, morphology, and how they ruined an obiting satellite. Tin whiskers grow in air or vacuum.
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==============================Original Headers============================== 32, 19 -- From smalinskas-at-yahoo.com Fri Jun 16 09:53:13 2006 32, 19 -- Received: from web34410.mail.mud.yahoo.com (web34410.mail.mud.yahoo.com [66.163.178.159]) 32, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5GErDcf012840 32, 19 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 09:53:13 -0500 32, 19 -- Received: (qmail 66812 invoked by uid 60001); 16 Jun 2006 14:53:12 -0000 32, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 32, 19 -- s=s1024; d=yahoo.com; 32, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 32, 19 -- b=ZaqnxzqR/WQm9UGzGSDQ9QDR2u7EqRGDp1gUTTsmNfHkKxWlQH1BwTvFTXVYpBXKX4SmcKwCyVc8BUbFNLgWPbdC7Ww1zCaT+83IV0/q39xwxFYWr3jIz33E5TV99VdjgCAJSXnlo9AnVLNU6uYPnQKuaZuNjlylNT6ruPENBMQ= ; 32, 19 -- Message-ID: {20060616145312.66810.qmail-at-web34410.mail.mud.yahoo.com} 32, 19 -- Received: from [141.151.33.213] by web34410.mail.mud.yahoo.com via HTTP; Fri, 16 Jun 2006 07:53:12 PDT 32, 19 -- Date: Fri, 16 Jun 2006 07:53:12 -0700 (PDT) 32, 19 -- From: Kestutis Smalinskas {smalinskas-at-yahoo.com} 32, 19 -- Subject: Re: [Microscopy] Tin Whiskers in high tin content solders. 32, 19 -- To: microscopy-at-ns.microscopy.com 32, 19 -- In-Reply-To: {200606152112.k5FLCRUK023781-at-ns.microscopy.com} 32, 19 -- MIME-Version: 1.0 32, 19 -- Content-Type: text/plain; charset=iso-8859-1 32, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I recall that there is a MT-2B in rm 110. What I don't remember is whether or not you have anywhere a *service* manual for one. We have the manuals that were shipped with the microtomes, even one old enough to be in a binder. We don't have a service manual. Do you have one, and if yes, can I get a photocopy of it? We have 3 of them, one horrible that's at best parts, and one I'm trying to resurrect before classes start this fall. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Fri Jun 16 12:34:22 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GHYMD1026600 3, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 12:34:22 -0500 3, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5GI9B4l003317 3, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 14:09:11 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Fri, 16 Jun 2006 13:34:22 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f0623090fc0b89b645ec8-at-[141.209.160.132]} 3, 20 -- Date: Fri, 16 Jun 2006 13:34:20 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: MT-2B 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 16 Jun 2006 17:34:22.0477 (UTC) FILETIME=[1ADBABD0:01C6916B] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -4 () L_EXCH_MF 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
If anyone out there has been doing any immunogold labeling for GFP, I would be interested in what primary you're using and how well it labels, depending on fixation.
Have a good weekend....
Lesley
Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322
==============================Original Headers============================== 8, 21 -- From lesley.bechtold-at-jax.org Fri Jun 16 12:47:10 2006 8, 21 -- Received: from carmen.jax.org (carmen.jax.org [192.43.249.16]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GHlAhu004528 8, 21 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 12:47:10 -0500 8, 21 -- Received: from jcs-mid-prod.jax.org (jcs-mid-prod.jax.org [192.43.249.134]) 8, 21 -- by carmen.jax.org (8.12.11/8.12.11) with ESMTP id k5GHP7ph004720 8, 21 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 13:47:08 -0400 (EDT) 8, 21 -- (envelope-from lesley.bechtold-at-jax.org) 8, 21 -- Received: from spikey.jax.org by jcs-mid-prod.jax.org 8, 21 -- with ESMTP id 99719541150479984; Fri, 16 Jun 2006 13:46:24 -0400 8, 21 -- From: "Lesley Bechtold" {lesley.bechtold-at-jax.org} 8, 21 -- To: "Microscopy Network" {microscopy-at-ns.microscopy.com} 8, 21 -- Subject: Immunogold Labeling for GFP 8, 21 -- Date: Fri, 16 Jun 2006 13:46:23 -0400 8, 21 -- Message-ID: {20060616134623491.00000004792-at-spikey} 8, 21 -- X-Mailer: Oracle Connector for Outlook 10.1.1.0.5 71011 (11.0.8010) 8, 21 -- X-Accept-Language: en-us, en 8, 21 -- MIME-Version: 1.0 8, 21 -- Content-Type: text/plain; charset=us-ascii 8, 21 -- Content-Transfer-Encoding: 8bit 8, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5GHlAhu004528 ==============================End of - Headers==============================
Sorry. My mousing finger glitched and picked the wrong name in my address book. Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 1, 20 -- From oshel1pe-at-cmich.edu Fri Jun 16 13:30:17 2006 1, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 1, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GIUHA5015534 1, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 13:30:17 -0500 1, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 1, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5GJ554l007634 1, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 15:05:05 -0400 1, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 1, 20 -- Fri, 16 Jun 2006 14:30:16 -0400 1, 20 -- Mime-Version: 1.0 1, 20 -- Message-Id: {f06230914c0b8a9028fd5-at-[141.209.160.132]} 1, 20 -- Date: Fri, 16 Jun 2006 14:30:14 -0400 1, 20 -- To: Microscopy-at-microscopy.com 1, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 1, 20 -- Subject: Oops MT-2B 1, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 1, 20 -- X-OriginalArrivalTime: 16 Jun 2006 18:30:16.0791 (UTC) FILETIME=[EA2F5E70:01C69172] 1, 20 -- X-CanItPRO-Stream: default 1, 20 -- X-Spam-Score: -4 () L_EXCH_MF 1, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
This is a well known problem that goes back to the early space programs. It has shown-up again since the "world" is switching to no-lead solders due the EU RoHS initiatives. Components cannot contain lead (Pb)in their lead (frame) finishes, so pure tin is the easiest solution. Unfortunately, component manufacturers are not aware of or forgot about the whisker problem also.
One of the most plausible scenarios was presented by Dr. Tu of UCLA at the 2003 ISTFA conference (See "Solder Failure Mechanisms, K.N. Tu, 29th International Symposium for Testing and Failure Analysis - Microelectronics Seminar, ASM International, 2003, P.187). He asserts that the whiskers grow in response to induced stresses in the tin plate deposit, caused by the volumetric changes due to Cu-Sn intermetallic compound formation. A nickel underplate slows this process due to slower Ni-Sn intermetallic compound growth kinetics.
For more info on tin whisker failures, go to http://nepp.nasa.gov/whisker/
Joseph
Joseph M. Oparowski Center for Materials Science Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
-----Original Message----- X-from: smalinskas-at-yahoo.com [mailto:smalinskas-at-yahoo.com] Sent: Friday, June 16, 2006 10:59 AM To: Oparowski, Joseph
Here are my thoughts:
If this phenomenon can be reliably reproduced, the effect from bias should be easily confirmed or denied.
This appears to be some type of transport mechanism... vapor transport, surface transport, bulk diffusion from the body of the metal, or grain boundary diffusion. It would be interesting to find where tin is being depleted in the solder joint, perhaps using EDS methods and metallographic cross sections. The challenge would be to distinguish tin depletion from normal solder casting segregation.
At the moment I'm assuming the whisker is a single crystal. Perhaps the whisker can be studied using x-ray crystallographic methods - either Laue or TEM methods. This may give a clue to the growth mechanism.
Stu Smalinskas, P.E. Metallurgist SKF Plymouth, Michigan
--- gary-at-gaugler.com wrote:
The whiskers are tin. Silver is added to lead solder to reduce the melting point.
Whiskering occurs at room temperature, high and low humidity and all sorts of conditions. It is a nasty problem for commercial and especially MIL applications. Alternate solders are being studied (different combinations of elements) but their melting point is sufficiently higher than Pb solder to be a real problem.
gary g.
---------------------------------------
At 01:35 PM 6/15/2006, you wrote:
Hi,
I would be curious to see any feedback on this subject, in the name of education.
I have a couple questions. Some of the lead-free solders have silver in them. Are you (or he) sure the whiskers are tin, or could they be silver? When you say "vacuum", are you referring to the low pressures in orbit, or a clean, high vacuum in a chamber of some sort? (or, perhaps, vacuum tube electronic component)
I believe the fields associated with a bias have something to do with it, but don't have any facts, or know what the mechanism is. I wonder if a high (strong) bias, in a vacuum chamber, would accelerate the phenomenon.
Looking forward to learning more. Thanks, Darrell
-----------------------------------------------
beaurega-at-westol.com wrote on 06/15/2006 10:55:59 AM:
Hi, I was asked to look into the morphology and cause of tin whiskers in high tin content lead free solders by a guy I meet. Some of the following questions have been asked for 50 years. Maybe someone out there can share some more recent expertise about tin whiskers.
* What is the root cause of crystalline tin whiskers? (It's probably not resolved yet. There are lots of conflicting opinions and observations on Google.)
* How do you prevent tin whiskers from growing out of almost pure tin solder joints? (He is not that interested in a thin film coating solution.)
* What is the mechanism(s) that can cause 2 micron diameter or less "fibers" (et al) to grow right out of the surface of solder joins to a length of up to one observed to be 10 mm long?
* What are the EDS results of tin whisker analysis?
* Are the EDS analysis results settled science?
* Have solder joints been ion milled to explore the interior of the solder or is this strictly a compression/stress surface phenomena? Or both?
He is NOT interested in high humidity dendritic growth of tin on PCB surfaces reputed to be caused by electric lines and associated equipotential lines. He is interested in the shorts caused by tin whiskers bridging solder joints through the air or a vacuum.
This raises the following big question. * How do you accelerate the growth of tin whiskers so that they can be studied without waiting 1-15 years for them to develop?
Please reply off list with any insights you have on this problem. I don't think this subject will have wide appeal. Thank you in advance for any perspectives that can be shared with him. If you are speculating on a solution or theory, please say so in your reply.
Paul
Disclaimers: I do not work for this guy's company, the guy, or any company or organization working on tin whiskers. I was astonished at the phenomenon, morphology, and how they ruined an obiting satellite. Tin whiskers grow in air or vacuum.
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 32, 19 -- From smalinskas-at-yahoo.com Fri Jun 16 09:53:13 2006 32, 19 -- Received: from web34410.mail.mud.yahoo.com (web34410.mail.mud.yahoo.com [66.163.178.159]) 32, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5GErDcf012840 32, 19 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 09:53:13 -0500 32, 19 -- Received: (qmail 66812 invoked by uid 60001); 16 Jun 2006 14:53:12 -0000 32, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 32, 19 -- s=s1024; d=yahoo.com; 32, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Cont ent-Type:Content-Transfer-Encoding; 32, 19 -- b=ZaqnxzqR/WQm9UGzGSDQ9QDR2u7EqRGDp1gUTTsmNfHkKxWlQH1BwTvFTXVYpBXKX4SmcK wCyVc8BUbFNLgWPbdC7Ww1zCaT+83IV0/q39xwxFYWr3jIz33E5TV99VdjgCAJSXnlo9AnVL NU6uYPnQKuaZuNjlylNT6ruPENBMQ= ; 32, 19 -- Message-ID: {20060616145312.66810.qmail-at-web34410.mail.mud.yahoo.com} 32, 19 -- Received: from [141.151.33.213] by web34410.mail.mud.yahoo.com via HTTP; Fri, 16 Jun 2006 07:53:12 PDT 32, 19 -- Date: Fri, 16 Jun 2006 07:53:12 -0700 (PDT) 32, 19 -- From: Kestutis Smalinskas {smalinskas-at-yahoo.com} 32, 19 -- Subject: Re: [Microscopy] Tin Whiskers in high tin content solders. 32, 19 -- To: microscopy-at-ns.microscopy.com 32, 19 -- In-Reply-To: {200606152112.k5FLCRUK023781-at-ns.microscopy.com} 32, 19 -- MIME-Version: 1.0 32, 19 -- Content-Type: text/plain; charset=iso-8859-1 32, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 46, 23 -- From Joseph_Oparowski-at-bose.com Fri Jun 16 13:33:00 2006 46, 23 -- Received: from BOSEMX02.bose.com (bosemx02.bose.com [139.68.146.21]) 46, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5GIWx1W019696 46, 23 -- for {Microscopy-at-Microscopy.Com} ; Fri, 16 Jun 2006 13:33:00 -0500 46, 23 -- Received: from USMAFREXMB02.bose.com ([139.68.132.238]) by USMAFREXCN02.bose.com with Microsoft SMTPSVC(6.0.3790.211); 46, 23 -- Fri, 16 Jun 2006 14:32:58 -0400 46, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 46, 23 -- Content-class: urn:content-classes:message 46, 23 -- MIME-Version: 1.0 46, 23 -- Content-Type: text/plain; 46, 23 -- charset="us-ascii" 46, 23 -- Subject: RE: [Microscopy] Re: Tin Whiskers in high tin content solders. 46, 23 -- Date: Fri, 16 Jun 2006 14:32:58 -0400 46, 23 -- Message-ID: {210C73AB4F6E0B4FBCB77B969C0BD84A029A7E2D-at-USMAFREXMB02.bose.com} 46, 23 -- X-MS-Has-Attach: 46, 23 -- X-MS-TNEF-Correlator: 46, 23 -- Thread-Topic: [Microscopy] Re: Tin Whiskers in high tin content solders. 46, 23 -- Thread-Index: AcaRVYq5+zcAKxcmTGi61vt0DqKzBAAGnAEA 46, 23 -- From: "Oparowski, Joseph" {Joseph_Oparowski-at-bose.com} 46, 23 -- To: {Microscopy-at-Microscopy.Com} 46, 23 -- X-OriginalArrivalTime: 16 Jun 2006 18:32:58.0282 (UTC) FILETIME=[4A70F0A0:01C69173] 46, 23 -- Content-Transfer-Encoding: 8bit 46, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5GIWx1W019696 ==============================End of - Headers==============================
We are starting a project looking at otolith microstructure, and I was wondering if folks have recommendations for mounting resins. We have one reference that recommends spurr's resin. Does anybody have experience with this type of preparation or a recommendation?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 4, 23 -- From hagglundk1-at-nku.edu Fri Jun 16 15:25:46 2006 4, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GKPkgG005767 4, 23 -- for {microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 15:25:46 -0500 4, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.1830); 4, 23 -- Fri, 16 Jun 2006 16:25:45 -0400 4, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 23 -- Content-class: urn:content-classes:message 4, 23 -- MIME-Version: 1.0 4, 23 -- Content-Type: text/plain; 4, 23 -- charset="US-ASCII" 4, 23 -- Subject: LM/SEM Mounting Otoliths 4, 23 -- Date: Fri, 16 Jun 2006 16:25:45 -0400 4, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F35875F-at-mailfac1.hh.nku.edu} 4, 23 -- X-MS-Has-Attach: 4, 23 -- X-MS-TNEF-Correlator: 4, 23 -- Thread-Topic: LM/SEM Mounting Otoliths 4, 23 -- Thread-Index: AcaRgwv65/QFS15YQtiOCGTvbfq4SQ== 4, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 4, 23 -- To: {microscopy-at-microscopy.com} 4, 23 -- X-OriginalArrivalTime: 16 Jun 2006 20:25:46.0084 (UTC) FILETIME=[0C5D9240:01C69183] 4, 23 -- Content-Transfer-Encoding: 8bit 4, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5GKPkgG005767 ==============================End of - Headers==============================
It's not too late to enter the International Metallographic Contest and Exhibit co-sponsored since 1972 by the International Metallographic Society and ASM International. The contest will be held in conjunction with the 39th annual technical meeting of the IMS and the M&M 2006 meeting this summer in Chicago. Twelve categories of competition. Best in show receives $3000 and the prestigious Jaquet-Lucas Award. Cash awards for first, second, and third place winners in eleven of the categories. Entries are prominently displayed during the M&M 2006 meeting and again in the fall during the ASM Annual Event. Deadline for entries is July 17. For additional information including rules, tips for creating a winning entry, judging guidelines, and examples of winning entries contact me or visit http://www.internationalmetallographicsociety.org/contest.html.
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 FAX: 508-699-4030
==============================Original Headers============================== 4, 21 -- From jeff-at-metallography.com Sat Jun 17 06:38:18 2006 4, 21 -- Received: from vw-cust-mail-app2-plano-126.emailscience.com (vw-cust-mail-app2-plano-126.emailscience.com [207.235.126.31]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5HBcI74012896 4, 21 -- for {Microscopy-at-microscopy.com} ; Sat, 17 Jun 2006 06:38:18 -0500 4, 21 -- Received: (qmail 95723 invoked by uid 1009); 17 Jun 2006 11:26:30 -0000 4, 21 -- Received: from unknown (HELO jstewart) (4.154.215.5) 4, 21 -- by vw-cust-mail-app2-plano.emailscience.com with SMTP; 17 Jun 2006 11:26:30 -0000 4, 21 -- Message-ID: {002001c69202$c801f840$959610ac-at-sternleach.com} 4, 21 -- Reply-To: "Jeff Stewart" {jeff-at-metallography.com} 4, 21 -- From: "Jeff Stewart" {jeff-at-metallography.com} 4, 21 -- To: {Microscopy-at-microscopy.com} 4, 21 -- Subject: International Metallographic Contest announcement 4, 21 -- Date: Sat, 17 Jun 2006 07:39:33 -0400 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: text/plain; 4, 21 -- charset="iso-8859-1" 4, 21 -- Content-Transfer-Encoding: 7bit 4, 21 -- X-Priority: 3 4, 21 -- X-MSMail-Priority: Normal 4, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1409 4, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 ==============================End of - Headers==============================
Our Hitachi S-4700 is suffering from 60 Hz EMF's in our lab. We see jagged lines in the horizontal direction at mags } /= to 150kX. I just had a vibration survey done and the results are within Hitachi specifications.
Do any of you pay particular attention to movement (of people, people in chairs, talking, etc.) when you are operating the SEM } 200kX? We pay particular attention to this in our HR-TEM lab. Just wondering if it is an issue in the FESEM lab too.
I'm looking for solutions. Cheap(er) DYI solutions. Any ideas?
Thanks!
OWen
Owen P. Mills Director, Materials Characterization & Fabrication Facilities Electron Optics Engineer, Applied Chemical & Morphological Analysis Laboratory
Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
==============================Original Headers============================== 12, 31 -- From opmills-at-mtu.edu Mon Jun 19 10:34:23 2006 12, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) 12, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JFYNfX014474 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 10:34:23 -0500 12, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 12, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id k5JFYNe6005289 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:23 -0400 12, 31 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 12, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k5JFYMjZ018584 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:22 -0400 12, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) 12, 31 -- by node34.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k5JFYLtJ011265 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:22 -0400 12, 31 -- (envelope-from opmills-at-mtu.edu) 12, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 12, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k5JFYLlX008693 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:21 -0400 (EDT) 12, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 12, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k5JFYLJ6022941 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:21 -0400 12, 31 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 31 -- Content-Transfer-Encoding: 7bit 12, 31 -- Message-Id: {F22556D6-9A4C-4046-94CF-A6BF69E16726-at-mtu.edu} 12, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 12, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 12, 31 -- Subject: SEM - electromagnetic fields 12, 31 -- Date: Mon, 19 Jun 2006 11:34:19 -0400 12, 31 -- X-Mailer: Apple Mail (2.750) 12, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.6.19.81433 12, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
I am sorry to say but round the world one of the problems that we all have is electromagnetic fields. To prove your problem compare working at a long WD with working at a very short WD. If the problem changes for the better at short WD it is a field problem. If it stays the same its most probably vibration or an instrument fault.
Check laboratories above, below and on all sides to track down the equipment that may be causing the problem, try switching equipment off.
As a warning to all, magnetic fields now prove to be the biggest FEG installation problem so always have a professional check your site. Remember the field will change by the inverse square so in some cases to move the instrument in the room may help. Field reduction systems work if a field is not over strong but the best route is to find a room which is truly field free.
Good luck
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {opmills-at-mtu.edu} To: {protrain-at-emcourses.com} Sent: Monday, June 19, 2006 4:37 PM
Hi All, I have been trying unsuccessfully to immuno-gold label HUVEC cells that have been embedded in LR White resin. The cells were plated and grown to supposed confluence, incubated with primary antibodies to PECAM and another adherence molecule, then fixed in 2% pfa, 0.5% glut., dehydrated, & embedded. The resin was cured at 50 deg. C overnight. En face sections were hydrated & blocked with buffer, then incubated with gold-tagged secondary Ab (anti-mouse or anti-rabbit). The sections were contrasted with aqueous Ur. Ac. then Pb citrate. I"ve seen nothing. For secondaries, I tried both F(ab)2s and whole IgG. I have been very successful with the same F(ab)2 secondaries on other samples processed the same way, but on which both primary and secondary Ab incubations were done on grid. In the past, we have successfully localized each of these primary Abs with peroxidase-DAB labelling prior to stringent fixation and embedding in Spurr's resin.
One of the problems is that HUVECs (human umbilical cord endothelial cells) are so darned flat...(less than 0.3 micrometers except for the nucleus) that its easy to cut right through them just trying to get a smooth section with no holes off of the raw block face. Another problem is that the proteins of interest are located at the cell junctions, in little vesicles and its not always easy to find cell-cell contacts.
My question (finally) is this: Has anyone tried to do an immuno-gold approach after immuno-peroxidase? We thought we'd like to try repeating the successful labelling of PECAM with DAB, pre-embedding, then try for the other Ab post-embedding.
Any ideas? (I'm sure there are....) thanks in advance, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 4, 21 -- From lcgould-at-med.cornell.edu Mon Jun 19 13:48:43 2006 4, 21 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JImg1C014989 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 13:48:43 -0500 4, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 4, 21 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5JImWJi025260 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 14:48:36 -0400 (EDT) 4, 21 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 4, 21 -- by mpx1.med.cornell.edu 4, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 4, 21 -- with ESMTPA id {0J1400HVGEWW1R30-at-mpx1.med.cornell.edu} for 4, 21 -- microscopy-at-microscopy.com; Mon, 19 Jun 2006 14:48:32 -0400 (EDT) 4, 21 -- Date: Mon, 19 Jun 2006 14:41:57 -0400 4, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 4, 21 -- Subject: EM: immuno labelling DAB & gold 4, 21 -- Sender: lcgould-at-med.cornell.edu 4, 21 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 4, 21 -- Message-id: {p0623091ac0bc9c974d8d-at-[140.251.48.23]} 4, 21 -- MIME-version: 1.0 4, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 4, 21 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.6.19.113432 ==============================End of - Headers==============================
1) I would worry that the DAB black mass would obscure the gold. I have not tried it, but it seems like this would be a problem. 2) Why not do primary and secondary Ab before fix, then fix hard and go into Epon? Your gold would be in the Epon. No post-cutting Ab work. 3) I would recommend cutting cross-sections of the cells. Then you do not have to worry about missing the flat cells when facing the block. Also, cell-cell junctions are much easier to see. I have protocols to do this if you want them.
David
_____________________
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
On Jun 19, 2006, at 11:54 AM, lcgould-at-med.cornell.edu wrote:
} } Hi All, } I have been trying unsuccessfully to immuno-gold label HUVEC cells } that have been embedded in LR White resin. The cells were plated and } grown to supposed confluence, incubated with primary antibodies to } PECAM and another adherence molecule, then fixed in 2% pfa, 0.5% } glut., dehydrated, & embedded. The resin was cured at 50 deg. C } overnight. En face sections were hydrated & blocked with buffer, } then incubated with gold-tagged secondary Ab (anti-mouse or } anti-rabbit). The sections were contrasted with aqueous Ur. Ac. then } Pb citrate. I"ve seen nothing. } For secondaries, I tried both F(ab)2s and whole IgG. I have been } very successful with the same F(ab)2 secondaries on other samples } processed the same way, but on which both primary and secondary Ab } incubations were done on grid. } In the past, we have successfully localized each of these primary Abs } with peroxidase-DAB labelling prior to stringent fixation and } embedding in Spurr's resin. } } One of the problems is that HUVECs (human umbilical cord endothelial } cells) are so darned flat...(less than 0.3 micrometers except for the } nucleus) that its easy to cut right through them just trying to get a } smooth section with no holes off of the raw block face. Another } problem is that the proteins of interest are located at the cell } junctions, in little vesicles and its not always easy to find } cell-cell contacts. } } My question (finally) is this: Has anyone tried to do an immuno-gold } approach after immuno-peroxidase? We thought we'd like to try } repeating the successful labelling of PECAM with DAB, pre-embedding, } then try for the other Ab post-embedding. } } Any ideas? (I'm sure there are....) } thanks in advance, } 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============================== 12, 22 -- From Elliott-at-arizona.edu Mon Jun 19 14:25:53 2006 12, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 12, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JJPrMx001845 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 14:25:53 -0500 12, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 12, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9E5D5E4B48E 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 12:25:52 -0700 (MST) 12, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 12, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 5748BE4B45B 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 12:25:51 -0700 (MST) 12, 22 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 22 -- In-Reply-To: {200606191854.k5JIsHFb022009-at-ns.microscopy.com} 12, 22 -- References: {200606191854.k5JIsHFb022009-at-ns.microscopy.com} 12, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 22 -- Message-Id: {6807EC57-D3D2-43A6-A1E0-8EEE3EED6C93-at-arizona.edu} 12, 22 -- Content-Transfer-Encoding: 7bit 12, 22 -- From: David Elliott {Elliott-at-arizona.edu} 12, 22 -- Subject: Re: [Microscopy] EM: immuno labelling DAB & gold 12, 22 -- Date: Mon, 19 Jun 2006 12:25:50 -0700 12, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 12, 22 -- X-Mailer: Apple Mail (2.750) 12, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
A friend wants to upgrade/keep going their Zeiss DSM 940A SEM.
First, they would like to know of any recommendations for a digital image acquisition upgrade available for this model. Contact off-list by commercial companies welcome.
Also, they are interested in anyone they can contact who might have the technical service manual, electrical schematics, etc., for this machine and/or spare parts that they might purchase or have donated. They are in Maine.
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============================== 8, 19 -- From tina-at-pbrc.hawaii.edu Mon Jun 19 16:35:13 2006 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JLZBUg023698 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 16:35:12 -0500 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k5JLZ6HC009676 8, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:06 -1000 (HST) 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k5JLZ5iU009673 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:05 -1000 (HST) 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 8, 19 -- Date: Mon, 19 Jun 2006 11:35:04 -1000 (HST) 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 8, 19 -- X-Sender: tina-at-halia 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 8, 19 -- Subject: Digital acquisition/parts for Zeiss DSM 940A 8, 19 -- Message-ID: {Pine.GSO.4.21.0606191129200.9568-100000-at-halia} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
My, isn't this a popular instrument to keep up and running... I have asked the same in the past few months...
Tell your friend in Maine to contact me, as I have been actively looking for replacement parts for this instrument, I have run across a fair amount of information. In fact, there is a group in Maine that has a Zeiss DM940A that they may be interested in getting to know....
Also, I would be interested in any information you may get from your request for digital image acquisition.
dj
On Mon, 19 Jun 2006, 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- } } A friend wants to upgrade/keep going their Zeiss DSM 940A SEM. } } First, they would like to know of any recommendations for a digital image } acquisition upgrade available for this model. Contact off-list by } commercial companies welcome. } } Also, they are interested in anyone they can contact who might have the } technical service manual, electrical schematics, etc., for this machine } and/or spare parts that they might purchase or have donated. They are in } Maine. } } 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============================== } 8, 19 -- From tina-at-pbrc.hawaii.edu Mon Jun 19 16:35:13 2006 } 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) } 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JLZBUg023698 } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 16:35:12 -0500 } 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) } 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k5JLZ6HC009676 } 8, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:06 -1000 (HST) } 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k5JLZ5iU009673 } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:05 -1000 (HST) } 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs } 8, 19 -- Date: Mon, 19 Jun 2006 11:35:04 -1000 (HST) } 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } 8, 19 -- X-Sender: tina-at-halia } 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} } 8, 19 -- Subject: Digital acquisition/parts for Zeiss DSM 940A } 8, 19 -- Message-ID: {Pine.GSO.4.21.0606191129200.9568-100000-at-halia} } 8, 19 -- MIME-Version: 1.0 } 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII } ==============================End of - Headers============================== }
==============================Original Headers============================== 8, 19 -- From dljones-at-bestweb.net Tue Jun 20 07:33:03 2006 8, 19 -- Received: from vms046pub.verizon.net (vms046pub.verizon.net [206.46.252.46]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5KCX3gS021164 8, 19 -- for {Microscopy-at-microscopy.com} ; Tue, 20 Jun 2006 07:33:03 -0500 8, 19 -- Received: from localhost ([71.249.29.149]) 8, 19 -- by vms046.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 8, 19 -- 9 2005)) with ESMTPA id {0J1500ES8S6LJ2CA-at-vms046.mailsrvcs.net} for 8, 19 -- Microscopy-at-microscopy.com; Tue, 20 Jun 2006 07:32:59 -0500 (CDT) 8, 19 -- Date: Tue, 20 Jun 2006 08:43:52 -0400 (Eastern Standard Time) 8, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 19 -- Subject: Re: [Microscopy] Digital acquisition/parts for Zeiss DSM 940A 8, 19 -- In-reply-to: {200606192141.k5JLfBEa030525-at-ns.microscopy.com} 8, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 19 -- To: tina-at-pbrc.hawaii.edu 8, 19 -- Cc: Microscopy-at-microscopy.com 8, 19 -- Message-id: {Pine.WNT.4.64.0606200837420.3168-at-H-F1} 8, 19 -- MIME-version: 1.0 8, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 19 -- References: {200606192141.k5JLfBEa030525-at-ns.microscopy.com} ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cloverbags-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 20, 2006 at 09:34:21 ---------------------------------------------------------------------------
Email: cloverbags-at-yahoo.com Name: raffaela
Organization: cebu doctors' university
Education: Undergraduate College
Location: Cebu City, Philippines,Asia
Question: what is the pH of a cedarwood oilJuniperus virginiana) for oil immersion objective? what is the pH of lemongrass oil(Cymbopogon citratus)?
Dear friends, please find the following CALL for PAPERS for Journal of Biomedical Optics All the best. Alby and Peri
---- May/June 2007
Visible Fluorescent Proteins
Guest Editors:
Alberto "Alby" Diaspro University of Genoa Department of Physics MicroScoBio-LAMBS/IFOM-NANOMED Via Dodecaneso 33 16146 Genova, Italy Tel: +39 10 353 6426 Fax: +39 10 314 218 E-mail: diaspro-at-fisica.unige.it
Ammasi Periasamy University of Virginia Keck Center for Cellular Imaging Biology Department Gilmer Hall (064), McCormick Rd Charlottesville, VA 22904 Tel: 434-243-7602 Fax: 434-982-5210 E-mail: ap3t-at-virginia.edu
Call for Papers: This special section will focus on the utilization of visible fluorescent proteins (VFPs) in fluorescence microscopy and spectroscopy. The use of fluorescent proteins and live-cell imaging has greatly improved our understanding of cell functioning in recent years. Discoveries in different organisms and development of fluorescent proteins are revolutionizing the study of cell behavior by providing convenient in vivo markers for gene expression and protein targeting in intact cells and organisms. VFPs coupled to three-dimensional fluorescence imaging methods such as confocal and multiphoton excitation microscopy are very effective in live-cell imaging. Moreover, the need to understand the fluorescence mechanism under different conditions has inspired a renaissance in single- molecule methods. Areas of interest also include papers related to fluorescence resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP) applications, VFP photophysical characterization, and photoconvertible and photactivatable mutants. In vivo applications are welcome as well as comparisons with potential competitors such as new fluorescent molecules and quantum dots.
Manuscripts due September 1, 2006
---------
--------------------------------------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001) --------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ----------------------------------------------
==============================Original Headers============================== 15, 31 -- From diaspro-at-fisica.unige.it Wed Jun 21 18:27:29 2006 15, 31 -- Received: from phobos.ge.infm.it (phobos.ge.infm.it [193.205.153.2]) 15, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5LNRSGJ009011 15, 31 -- for {microscopy-at-microscopy.com} ; Wed, 21 Jun 2006 18:27:28 -0500 15, 31 -- Received: from [193.205.153.216] (albypc.ge.infm.it [193.205.153.216]) 15, 31 -- by phobos.ge.infm.it (8.11.6/8.11.0) with ESMTP id k5L6ZXh10248; 15, 31 -- Wed, 21 Jun 2006 08:35:33 +0200 15, 31 -- Mime-Version: 1.0 (Apple Message framework v750) 15, 31 -- Content-Transfer-Encoding: 7bit 15, 31 -- Message-Id: {85421244-1096-48EB-89F8-934BB7B5F916-at-fisica.unige.it} 15, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 15, 31 -- To: mplsm-users-at-yahoogroups.com, 15, 31 -- Confocal List Microscopy {CONFOCAL-at-LISTSERV.BUFFALO.EDU} , 15, 31 -- microscopy-at-microscopy.com, Ronzitti Emiliano {ronzi81-at-virgilio.it} , 15, 31 -- Cella Francesca {fra_cella-at-yahoo.it} , 15, 31 -- Mondal Partha Pratim {partha-at-fisica.unige.it} , 15, 31 -- Magrassi Raffaella {magrassi-at-fisica.unige.it} , 15, 31 -- Caorsi Valentina {caorsi-at-fisica.unige.it} , 15, 31 -- Diaspro Alberto {diaspro-at-fisica.unige.it} , 15, 31 -- Bianchini Paolo {bianchini-at-ge.infm.it} , 15, 31 -- Gattodub Gattodub {gattodub-at-email.it} , 15, 31 -- Mazza Davide {mazza-at-fisica.unige.it} , 15, 31 -- Morotti Federica {morottina-at-yahoo.it} , 15, 31 -- Testa Ilaria {ilaria.testa-at-email.it} , 15, 31 -- Vicidomini Giuseppe {vicidomini-at-fisica.unige.it} , 15, 31 -- Testa Ilaria {testa-at-fisica.unige.it} , 15, 31 -- Mario Faretta {mario.faretta-at-ifom-ieo-campus.it} 15, 31 -- From: diaspro {diaspro-at-fisica.unige.it} 15, 31 -- Subject: JBO special issue - call for papers 15, 31 -- Date: Wed, 21 Jun 2006 08:30:47 +0200 15, 31 -- X-Mailer: Apple Mail (2.750) ==============================End of - Headers==============================
A enthusiastic construction worker cut a major telecom trunk line out here in the suburbs of Chicago a little over 24 hours ago. Took out ALL telecom links, phone lines, networks the whole system. It made for a very quiet and annoying 24 hours here. Amazing how rapidly wedded we become to communications links.
Anyway, although the Listserver was running it was not able to "talk to anyone".
If you attempted posting anything during the las 24 hours there is a good likelihood it was held in queue somewhere on the net and it may get resent. If your not sure then feel free to repost any messages.
Cheers...
Nestor Your Friendly Neighborhood SysOp (who had a very quiet 24 + hours )
==============================Original Headers============================== 9, 11 -- From zaluzec-at-microscopy.com Wed Jun 21 19:57:21 2006 9, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 9, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5M0vJXJ023886 9, 11 -- for {microscopy-at-microscopy.com} ; Wed, 21 Jun 2006 19:57:20 -0500 9, 11 -- Mime-Version: 1.0 9, 11 -- Message-Id: {p06110402c0bf988fd43d-at-[206.69.208.22]} 9, 11 -- Date: Wed, 21 Jun 2006 19:57:19 -0500 9, 11 -- To: microscopy-at-microscopy.com 9, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 9, 11 -- Subject: Administrivia: Network Connections have been restored 9, 11 -- 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://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both kjl226-at-vt.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: kjl226-at-vt.edu Name: Kathy
Organization: Virginia Tech
Title-Subject: [Filtered] Salary ranges
Question: What is a fair salary for a supervisory position, where the person will run an Electron Microscope Service Laboratory? This lab will offer all phases of processing of SEM amd TEM samples. Sectioning of the TEM samples, thick and/or thins, negative staining. Running both scopes and assist with use of the scopes including capturing digital images.
This person will also be responible for ordering all supplies, undating SOP's as needed, keeping labortories maintained, helping students with projects, keeping inventory of equipment, keeping all equipment in good working condition, calling service personnel when scopes on service contracts need work, and calulating bills for EM services.
Check out the Microscopy Today archives for the salary survey done in 2004 by Ron Anderson and Barb Foster. It contains a lot of valuable comparison data that should help you determine an appropriate salary range for your geographical location and responsibilities.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
On 6/21/06 9:04 PM, "kjl226-at-vt.edu" {kjl226-at-vt.edu} wrote:
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==============================Original Headers============================== 7, 22 -- From dsherman-at-purdue.edu Wed Jun 21 20:46:36 2006 7, 22 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5M1kaBE012448 7, 22 -- for {microscopy-at-microscopy.com} ; Wed, 21 Jun 2006 20:46:36 -0500 7, 22 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 22 -- Wed, 21 Jun 2006 21:46:35 -0400 7, 22 -- Received: from 74.132.211.81 ([74.132.211.81]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 7, 22 -- Thu, 22 Jun 2006 01:46:34 +0000 7, 22 -- User-Agent: Microsoft-Entourage/11.2.3.060209 7, 22 -- Date: Wed, 21 Jun 2006 21:46:37 -0400 7, 22 -- Subject: Re: [Microscopy] viaWWW: Salary ranges 7, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 22 -- To: {kjl226-at-vt.edu} , "message to: MSA list" {microscopy-at-microscopy.com} 7, 22 -- Message-ID: {C0BF6EBD.37F0%dsherman-at-purdue.edu} 7, 22 -- Thread-Topic: [Microscopy] viaWWW: Salary ranges 7, 22 -- Thread-Index: AcaVnbLe8ZosfgGQEduvUQAKlcoUxg== 7, 22 -- In-Reply-To: {200606220104.k5M14nlM005892-at-ns.microscopy.com} 7, 22 -- Mime-version: 1.0 7, 22 -- Content-type: text/plain; 7, 22 -- charset="US-ASCII" 7, 22 -- Content-transfer-encoding: 7bit 7, 22 -- X-OriginalArrivalTime: 22 Jun 2006 01:46:35.0710 (UTC) FILETIME=[B21A25E0:01C6959D] ==============================End of - Headers==============================
Question: what is the pH of a cedarwood oilJuniperus virginiana) for oil immersion objective? what is the pH of lemongrass oil(Cymbopogon citratus)?
1. There are three types of cedar oil:
Virginia cedarwood oil (Juniperus virginiana), Texas cedarwood oil (Juniperus ashei or mexicana), and Western red cedar (Thuja plicata)
I have cedar oil (Texas) from Polysciences. Their MSDS does not list a pH. Nor does Merck list one. The Sigma Aldrich MSDS does not list one, nor does the Libety Nautral Products (Virginia), nor does JT Baker, nor does Well, Naturally Products (Virginia & Texas & Western).
I will try to get a pH on my Texas cedar oil next week and report this at least.
The constituents are mainly cedrene (a terpene) and cedral (cedar camphor).
2. regarding lemongrass oil:
I have found no pH. Well, Naturally Products does not have it in their MSDS.
3. I have a client that extracts, sells, distributes these oils and will check for their data as well next week.
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
This message is from pH2. This message and any attachments may contain legally privileged or confidential information, and are intended only for the individual or entity identified above as the addressee. If you are not the addressee, or if this message has been addressed to you in error, you are not authorized to read, copy, or distribute this message and any attachments, and we ask that you please delete this message and attachments (including all copies) and notify the sender by return e-mail or by phone at 317-752-6386. Delivery of this message and any attachments to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or a privilege. All personal messages express views only of the sender, which are not to be attributed to pH2 and may not be copied or distributed without this statement.
==============================Original Headers============================== 21, 32 -- From ph2-at-sprynet.com Wed Jun 21 22:34:37 2006 21, 32 -- Received: from elasmtp-junco.atl.sa.earthlink.net (elasmtp-junco.atl.sa.earthlink.net [209.86.89.63]) 21, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5M3Yano024776 21, 32 -- for {Microscopy-at-Microscopy.Com} ; Wed, 21 Jun 2006 22:34:37 -0500 21, 32 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 21, 32 -- s=dk20050327; d=sprynet.com; 21, 32 -- b=Xd/a6Vln7K7YcDyDRjxr7rahw0V65rGdYJkVgR8/2tw4AvfSEPsCF2sUI+xxbFg6; 21, 32 -- h=Received:Reply-To:From:To:Cc:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; 21, 32 -- Received: from [4.224.249.140] (helo=yourw92p4bhlzg) 21, 32 -- by elasmtp-junco.atl.sa.earthlink.net with asmtp (TLSv1:RC4-MD5:128) 21, 32 -- (Exim 4.34) 21, 32 -- id 1FtFxi-0004ct-Pf; Wed, 21 Jun 2006 23:34:36 -0400 21, 32 -- Reply-To: {ph2-at-sprynet.com} 21, 32 -- From: "Tony Havics" {ph2-at-sprynet.com} 21, 32 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} 21, 32 -- Cc: {cloverbags-at-yahoo.com} 21, 32 -- Subject: pH of Oils 21, 32 -- Date: Wed, 21 Jun 2006 23:39:37 -0500 21, 32 -- Organization: pH2 21, 32 -- Message-ID: {002901c695b5$e0e0dcd0$8cf9e004-at-QEPI.local} 21, 32 -- MIME-Version: 1.0 21, 32 -- Content-Type: text/plain; 21, 32 -- charset="UTF-8" 21, 32 -- X-Priority: 3 (Normal) 21, 32 -- X-MSMail-Priority: Normal 21, 32 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 21, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 21, 32 -- Importance: Normal 21, 32 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f9bccc22a0f0e71d8e2dcc2d4da497cf5a350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 21, 32 -- X-Originating-IP: 4.224.249.140 21, 32 -- Content-Transfer-Encoding: 8bit 21, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5M3Yano024776 ==============================End of - Headers==============================
Greetings, It is likely that the pH of any oil is 7, almost by definition. I am sure a real chemist will correct me but pH expresses the relative abundance of protons over hydroxyls. As oils are non polar, they presumably repel a positive and negative charge equally and hence the pH is 'neutral', or a value of 7. Furthermore, the whole pH system is developed for aqueous solutions and there is presumably very little water indeed in the cedar oil, so it may not even be sensible to speak of pH for oils. Presumably that is why you can't find values in MSDSs, ehgo. Perhaps there is some solid-state notion of 'proton activity' that applies? Again maybe a chemist or physicist will have a more rigorous answer. Why do you care about the pH of these oils?
Hope this helps, Tobias } } Regarding: } } mail: cloverbags-at-yahoo.com } Name: raffaela } } Organization: cebu doctors' university } } Education: Undergraduate College } } Location: Cebu City, Philippines,Asia } } Question: what is the pH of a cedarwood } oilJuniperus virginiana) for oil immersion } objective? what is the pH of lemongrass } oil(Cymbopogon citratus)? } } 1. There are three types of cedar oil: } } Virginia cedarwood oil (Juniperus virginiana), } Texas cedarwood oil (Juniperus ashei or } mexicana), and Western red cedar (Thuja plicata) } } I have cedar oil (Texas) from Polysciences. } Their MSDS does not list a pH. Nor does Merck } list one. The Sigma Aldrich MSDS does not list } one, nor does the Libety Nautral Products } (Virginia), nor does JT Baker, nor does Well, } Naturally Products (Virginia & Texas & Western). } } I will try to get a pH on my Texas cedar oil } next week and report this at least. } } The constituents are mainly cedrene (a terpene) and cedral (cedar camphor). } } 2. regarding lemongrass oil: } } I have found no pH. Well, Naturally Products does not have it in their MSDS. } } Constituents (as listed by Merck): 75-85% } citral, methylheptenone, citronellel, geraniol, } limonene } } 3. I have a client that extracts, sells, } distributes these oils and will check for their } data as well next week. } } Tony } } .......................................................................... } Andrew Anthony "Tony" Havics, CHMM, CIH, PE } pH2, LLC } PO Box 34140 } Indianapolis, IN 46234 } (317) 752-6386 } (317) 409-3238 cell } } 90% of Risk Management is knowing where to place } the decimal point...any consultant can give you } the other 10%’ŃÝ } } This message is from pH2. This message and any } attachments may contain legally privileged or } confidential information, and are intended only } for the individual or entity identified above as } the addressee. If you are not the addressee, or } if this message has been addressed to you in } error, you are not authorized to read, copy, or } distribute this message and any attachments, and } we ask that you please delete this message and } attachments (including all copies) and notify } the sender by return e-mail or by phone at } 317-752-6386. Delivery of this message and any } attachments to any person other than the } intended recipient(s) is not intended in any way } to waive confidentiality or a privilege. All } personal messages express views only of the } sender, which are not to be attributed to pH2 } and may not be copied or distributed without } this statement. } } } } ==============================Original Headers============================== } 21, 32 -- From ph2-at-sprynet.com Wed Jun 21 22:34:37 2006 } 21, 32 -- Received: from } elasmtp-junco.atl.sa.earthlink.net } (elasmtp-junco.atl.sa.earthlink.net } [209.86.89.63]) } 21, 32 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k5M3Yano024776 } 21, 32 -- for {Microscopy-at-Microscopy.Com} ; } Wed, 21 Jun 2006 22:34:37 -0500 } 21, 32 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 21, 32 -- s=dk20050327; d=sprynet.com; } 21, 32 -- } b=Xd/a6Vln7K7YcDyDRjxr7rahw0V65rGdYJkVgR8/2tw4AvfSEPsCF2sUI+xxbFg6; } 21, 32 -- } h=Received:Reply-To:From:To:Cc:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; } 21, 32 -- Received: from [4.224.249.140] (helo=yourw92p4bhlzg) } 21, 32 -- by } elasmtp-junco.atl.sa.earthlink.net with asmtp } (TLSv1:RC4-MD5:128) } 21, 32 -- (Exim 4.34) } 21, 32 -- id 1FtFxi-0004ct-Pf; Wed, 21 Jun 2006 23:34:36 -0400 } 21, 32 -- Reply-To: {ph2-at-sprynet.com} } 21, 32 -- From: "Tony Havics" {ph2-at-sprynet.com} } 21, 32 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} } 21, 32 -- Cc: {cloverbags-at-yahoo.com} } 21, 32 -- Subject: pH of Oils } 21, 32 -- Date: Wed, 21 Jun 2006 23:39:37 -0500 } 21, 32 -- Organization: pH2 } 21, 32 -- Message-ID: {002901c695b5$e0e0dcd0$8cf9e004-at-QEPI.local} } 21, 32 -- MIME-Version: 1.0 } 21, 32 -- Content-Type: text/plain; } 21, 32 -- charset="UTF-8" } 21, 32 -- X-Priority: 3 (Normal) } 21, 32 -- X-MSMail-Priority: Normal } 21, 32 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 } 21, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 21, 32 -- Importance: Normal } 21, 32 -- X-ELNK-Trace: } 6888e50b2be9b4fee5331016acda17f9bccc22a0f0e71d8e2dcc2d4da497cf5a350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c } 21, 32 -- X-Originating-IP: 4.224.249.140 } 21, 32 -- Content-Transfer-Encoding: 8bit } 21, 32 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } k5M3Yano024776 } ==============================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 rpowell-at-nanoprobes.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Not sure how flexible you are in terms of your staining and preparation procedure... a PubMed search with the terms "post-embedding AND gold AND peroxidase" gave me 35 results, citing a variety of combinations of methods.
We have encountered several papers where silver-enhanced Nanogold (disclaimer - Nanogold is our product!) was used with DAB for double, or in the most recent case triple immunolabeling. We have summarized a couple of these in our newsletter - here are the references and links:
(1) Triple labeling, pre-embedding procedure:
Feng, Y.-P.; Li, Y.-Q.; Wang, W.; Wu, S.-X.; Chen, T.; Shigemoto, R., and Mizuno, N.: Morphological evidence for GABA/glycine-cocontaining terminals in synaptic contact with neurokinin-1 receptor-expressing neurons in the sacral dorsal commissural nucleus of the rat. Neurosci. Lett., 18, 144-148.
Reference describing methods in more detail:
Li, J. L.; Wang, D.; Kaneko, T.; Shigemoto, R.; Nomura, S., and Mizuno, N.: Relationship between neurokinin-1 receptor and substance P in the striatum: light and electron microscopic immunohistochemical study in the rat. J. Comp. Neurol., 418, 156ń163 (2000).
Our article:
http://www.nanoprobes.com/Vol6_Iss11.html#3
(2) Pre-embedding localization of m2R and either ChAT or VAChT in the nucleus basalis magnocellularis (NBM) in rat brain sections where m2R was detected using pre-embedding ultrasmall (0.8 nm) gold enhanced with HQ Silver, and ChAT or VAChT by pre-embedding immunoperoxidase using the peroxidase anti-peroxidase (PAP) technique:
Decossas, M.; Doudnikoff, E.; Bloch, B., and Bernard, V.: Aging and subcellular localization of m2 muscarinic autoreceptor in basalocortical neurons in vivo. Neurobiol. Aging, 26, 1061-1072 (2005).
Our article:
http://www.nanoprobes.com/Vol6_Iss3.html#5
(3) Similar (pre-embedding labeling of vesicular glutamate transporter 1 (VGLUT1) and choline acetyltransferase (ChAT) immuno-reactivity respectively in the rat lumbar spinal cord):
Wu, S.-X.; Koshimizu, Y.; Feng, Y. P.; Okamoto, K.; Fujiyama, F.; Hioki, H.; Li, Y. Q.; Kaneko, T., and Mizuno, N.: Vesicular glutamate transporter immunoreactivity in the central and peripheral endings of muscle-spindle afferents. Brain Res.,} 1011, 247-251 (2004).
Other pre-embedding method:
Gutekunst, C. A.; Torre, E. R.; Sheng, Z.; Yi, H.; Coleman, S. H.; Riedel, I. B., and Bujo, H.: Stigmoid Bodies Contain Type I Receptor Proteins SorLA/LR11 and Sortilin. New perspectives on their function. J. Histochem. Cytochem., 51, 841-852 (2003).
Our article:
http://www.nanoprobes.com/Vol5_Iss7.html#4
Hope some of these are helpful,
Rick Powell Nanoprobes, Incorporated www.nanoprobes.com
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Hi All, I have been trying unsuccessfully to immuno-gold label HUVEC cells that have been embedded in LR White resin. The cells were plated and grown to supposed confluence, incubated with primary antibodies to PECAM and another adherence molecule, then fixed in 2% pfa, 0.5% glut., dehydrated, & embedded. The resin was cured at 50 deg. C overnight. En face sections were hydrated & blocked with buffer, then incubated with gold-tagged secondary Ab (anti-mouse or anti-rabbit). The sections were contrasted with aqueous Ur. Ac. then Pb citrate. I"ve seen nothing. For secondaries, I tried both F(ab)2s and whole IgG. I have been very successful with the same F(ab)2 secondaries on other samples processed the same way, but on which both primary and secondary Ab incubations were done on grid. In the past, we have successfully localized each of these primary Abs with peroxidase-DAB labelling prior to stringent fixation and embedding in Spurr's resin.
One of the problems is that HUVECs (human umbilical cord endothelial cells) are so darned flat...(less than 0.3 micrometers except for the nucleus) that its easy to cut right through them just trying to get a smooth section with no holes off of the raw block face. Another problem is that the proteins of interest are located at the cell junctions, in little vesicles and its not always easy to find cell-cell contacts.
My question (finally) is this: Has anyone tried to do an immuno-gold approach after immuno-peroxidase? We thought we'd like to try repeating the successful labelling of PECAM with DAB, pre-embedding, then try for the other Ab post-embedding.
Any ideas? (I'm sure there are....) thanks in advance, Lee
The histotechnologist in our group is having difficulty keeping formalin-fixed, paraffin-embedded sections of pig hooves on the slide for staining. Does anyone have any suggestions for her? She uses superfrost slides which works for all other tissues. If you have any hints for changes in fixation, etc. please pass those along.
Thanks! Peggy Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
==============================Original Headers============================== 6, 23 -- From MSHERWOOD-at-PARTNERS.ORG Thu Jun 22 09:31:02 2006 6, 23 -- Received: from PHSXCON5.partners.org (phsxcon5.mgh.harvard.edu [132.183.130.38]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MEV2ls001555 6, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 09:31:02 -0500 6, 23 -- Received: from PHSXMB1.partners.org ([132.183.118.117]) by PHSXCON5.partners.org with Microsoft SMTPSVC(6.0.3790.211); 6, 23 -- Thu, 22 Jun 2006 10:31:00 -0400 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="iso-8859-1" 6, 23 -- Subject: FW: [LM Microsocpy] Re: staining of animal hooves and/or nails 6, 23 -- Date: Thu, 22 Jun 2006 10:31:00 -0400 6, 23 -- Message-ID: {1AF23D0AD12E7444A5DB083CA978B73407A309DD-at-PHSXMB1.partners.org} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [LM Microsocpy] Re: staining of animal hooves and/or nails 6, 23 -- Thread-Index: AcaWCBNPN2zboBv3S4eAdaMG2lZvqwAAFWkQ 6, 23 -- From: "Sherwood, Margaret " {MSHERWOOD-at-PARTNERS.ORG} 6, 23 -- To: {Microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 22 Jun 2006 14:31:00.0271 (UTC) FILETIME=[7B82C3F0:01C69608] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MEV2ls001555 ==============================End of - Headers==============================
I was under the impression that, although oils can contain acidic contaminates, these are often removed during the refining process (not being good for engines etc.). Acidity (the acid-type constituents) "is usually defined in terms of total acid number (TAN). These constituents vary in nature and may or may not markedly influence the behaviour of the lubricant". Low acidity oils can oxidise during use and become more acidic over time (years) - e.g. the acid number goes to over 0.4 - while the newly refined 'non-corrosive' oil would be expected to be nearer to 0.05. The acids can be weak organic and strong inorganic. Acidity in oil can be useful for things like improved water resistance. Oils don't have an aqueous pH as such though, just the TAN number (total acidic number).
Olive oil for example is also graded for consumption on it's oleic acid acidity - around 3% in cheaper olive oil, and down to below 1% in 'extra virgin'. Oils are generally insoluble in water, which rather helps reduce it's corrosive properties during use.
Cargille use the Neutralization Number for their immersion oils. This is a measure of the acidity or alkalinity of the oil and is the mass in milligrams of the amount of acid (HCl) or base (KOH) required to neutralize one gram of the oil. For their standard immersion oils and type FF the value is very low [acidity] at 0.01 [KOH] (whereas the type DF is a rather high at 0.15 and the bottle states that it may damage objectives in constant use - it has lower auto-fluorescence than FF though). Cargille states that Cedarwood oil has a KOH neutralisation number greater than these values (ie. it is more acidic).
When used for objective immersion, the main things of interest in an oil would be things like : refractive index, colour, fluorescent properties, likely damage to the lens cement, crystallisation when cold (reversible), and whether the oil evaporates leaving a hard residue. The metal outside parts of the objective generally have an anti-corrosion finish, although here with our inverted microscopes spillage of corrosive culture media has sometimes caused some damage at the base of the objective, in particular corrosion of the area around the screw thread. The immersion oils generally dissolve in things like ethyl alcohol. Rather like engine oils for your car one tends to stick with the manufacturers [typically expensive] recommended one and not look too deeply into it's chemistry (microscope manufacturer's will soak test their objectives immersed for 6 months or more in an immersion oil before recommending it).
Oil acids can attack the lens mounting cement [see Cargille's comments below]. Plus the organic solvents used to remove lens immersion oil (like Xylene and ethanol) have been implicated in lens cement damage - I use ether generally for cleaning (and only clean rarely, when the lens is too dirty to be cleaned by just a lens tissue). Modern [low acid content] immersion oils supplied by the microscope manufacturer seem to be safe for longer term contact with the objective and don't evaporate to a hard residue (unlike say corn oil).
Cargille quote immersion oil shelf lives of 10 years (FF) and 5 years (DF) - and half that when opened.
On Cargille's site [http://www.cargille.com/immersionoilmicroscope.shtml] John Cargille states:
"To the knowledge of the author, no immersion oil matches the optics of the microscope perfectly. One of the closest matches is thickened cedarwood oil which, for many years, was the most widely used, if not the only immersion oil available. The disadvantageous properties of cedarwood oil are: high absorption of blue and UV light, yellowing with age, a tendency to harden on lenses due to uneven volatility, acidity, and changing viscosity (diluting with solvent changes the index and dispersion). The synthetic immersion oils eliminate many of the disadvantages cited above.
The acid value of immersion oil should be very low. The synthetics usually have acid values lower than cedarwood oil.
High acidity can, in time, affect the condition of the metal parts of the objective, and possibly more important, can cause deterioration of lens cements. The lens cement is also a seal that prevents oil from penetrating to the back of the lens. A crack or perforation in the cement draws oil by capillary attraction and a thin film of oil slowly creeps over the back of the objective lens; a poor image may develop without being immediately noticed. When the microscopist realizes the image has deteriorated, he may not realize the cause unless he has faced this problem before."
See Cargille's excellent site for more immersion oil info.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: ph2-at-sprynet.com [mailto:ph2-at-sprynet.com] Sent: 22 June 2006 04:40 To: keith.morris-at-ucl.ac.uk
Regarding:
mail: cloverbags-at-yahoo.com Name: raffaela
Organization: cebu doctors' university
Education: Undergraduate College
Location: Cebu City, Philippines,Asia
Question: what is the pH of a cedarwood oilJuniperus virginiana) for oil immersion objective? what is the pH of lemongrass oil(Cymbopogon citratus)?
1. There are three types of cedar oil:
Virginia cedarwood oil (Juniperus virginiana), Texas cedarwood oil (Juniperus ashei or mexicana), and Western red cedar (Thuja plicata)
I have cedar oil (Texas) from Polysciences. Their MSDS does not list a pH. Nor does Merck list one. The Sigma Aldrich MSDS does not list one, nor does the Libety Nautral Products (Virginia), nor does JT Baker, nor does Well, Naturally Products (Virginia & Texas & Western).
I will try to get a pH on my Texas cedar oil next week and report this at least.
The constituents are mainly cedrene (a terpene) and cedral (cedar camphor).
2. regarding lemongrass oil:
I have found no pH. Well, Naturally Products does not have it in their MSDS.
3. I have a client that extracts, sells, distributes these oils and will check for their data as well next week.
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
==============================Original Headers============================== 43, 27 -- From keith.morris-at-ucl.ac.uk Thu Jun 22 09:52:39 2006 43, 27 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) 43, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MEqdvX011845 43, 27 -- for {Microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 09:52:39 -0500 43, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 43, 27 -- by vscani-d.ucl.ac.uk with esmtp (Exim 4.60) 43, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 43, 27 -- id 1FtQXs-0002j4-SV; Thu, 22 Jun 2006 15:52:37 +0100 43, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 43, 27 -- To: {Microscopy-at-microscopy.com} 43, 27 -- Cc: {cloverbags-at-yahoo.com} 43, 27 -- Subject: RE: [Microscopy] pH of Oils 43, 27 -- Date: Thu, 22 Jun 2006 15:52:33 +0100 43, 27 -- Message-ID: {000001c6960b$7ea22500$7b865290-at-keithhigrade} 43, 27 -- MIME-Version: 1.0 43, 27 -- Content-Type: text/plain; 43, 27 -- charset="iso-8859-1" 43, 27 -- X-Mailer: Microsoft Office Outlook 11 43, 27 -- Thread-Index: AcaVrZneujlpeoxMTIOEKedyM/LyNwATipAg 43, 27 -- In-Reply-To: {200606220340.k5M3eN8d031970-at-ns.microscopy.com} 43, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 43, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 43, 27 -- X-UCL-MailScanner: Found to be clean 43, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 43, 27 -- X-Spam-Status: No 43, 27 -- Content-Transfer-Encoding: 8bit 43, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MEqdvX011845 ==============================End of - Headers==============================
We have always used ethanol as the dehydration solvent with our Balzers CPD 020. Does anyone know whether the seals and other components of this instrument are compatible with methanol? It is used as a primary fixative for plant tissue in a publication, but it is unclear from the methods section whether the tissue is changed to ethanol before critical point drying. We have sent e-mails to the author and US supplier of Balzers equipment, but not yet received any replies. Also, does anyone know whether absolute ethanol can be directly changed for anhydrous methanol without damaging plant tissue?
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall 3209 N. Maryland Ave. Milwaukee, WI 53211 USA
Phone: (414)229-6816
==============================Original Headers============================== 7, 19 -- From owenha-at-csd.uwm.edu Thu Jun 22 11:33:17 2006 7, 19 -- Received: from batch3.csd.uwm.edu (batch3.csd.uwm.edu [129.89.169.226]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MGXGdi023787 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 11:33:17 -0500 7, 19 -- Received: from alpha2.csd.uwm.edu (alpha2.csd.uwm.edu [129.89.7.202] (may be forged)) 7, 19 -- by batch3.csd.uwm.edu (8.13.6/8.12.6) with ESMTP id k5MGXEav030866 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 11:33:14 -0500 (CDT) 7, 19 -- Received: from localhost (owenha-at-localhost) 7, 19 -- by alpha2.csd.uwm.edu (8.13.6/8.12.6) with SMTP id k5MGXEOd004137 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 11:33:14 -0500 (CDT) 7, 19 -- X-Authentication-Warning: alpha2.csd.uwm.edu: owenha owned process doing -bs 7, 19 -- Date: Thu, 22 Jun 2006 11:33:14 -0500 (CDT) 7, 19 -- From: Heather A Owen {owenha-at-csd.uwm.edu} 7, 19 -- Reply-To: Heather A Owen {owenha-at-csd.uwm.edu} 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- Subject: Balzers Critical Point Drier solvents 7, 19 -- Message-ID: {Pine.OSF.3.96.1060622111531.10763A-100000-at-alpha2.csd.uwm.edu} 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
We have been polymerizing our excess resins from infiltrations, etc., in the same oven we use for polymerizing our actual samples in resin. My question is whether this could have an effect on the quality of the resin blocks, maybe due to the solvents in the leftovers?
We had a couple batches of semi-gummy bears recently and this question has been resinating around the lab ever since. No samples lost, but a definite annoyance.
Thanks for any thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jun 22 12:52:34 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MHqYXe002752 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 12:52:34 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: TEM: polymerization question 10, 23 -- Date: Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E332E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: TEM: polymerization question 10, 23 -- thread-index: AcaWJKO/b+LSDKGdS4WFWVdZe1OTcA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 22 Jun 2006 17:52:34.0055 (UTC) FILETIME=[A3F7C170:01C69624] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MHqYXe002752 ==============================End of - Headers==============================
First a question: Is the oven in a hood? And is the oven ventilated on top (most of them I've run across either have pluggable holes or some sort of vane type shutter on top.
I haven't noticed any issue polymerizing with alcohol contaminated resin in the oven at the same time as the blocks. Although more often than not I will let the waste sit out in the hood until the blocks are cured and then stick in the waste to cure until next time I'm ready to put a batch in.
HTH
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Thursday, June 22, 2006 1:57 PM To: Williams, Geoffrey
Hi all,
We have been polymerizing our excess resins from infiltrations, etc., in the same oven we use for polymerizing our actual samples in resin. My question is whether this could have an effect on the quality of the resin blocks, maybe due to the solvents in the leftovers?
We had a couple batches of semi-gummy bears recently and this question has been resinating around the lab ever since. No samples lost, but a definite annoyance.
Thanks for any thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jun 22 12:52:34 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MHqYXe002752 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 12:52:34 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: TEM: polymerization question 10, 23 -- Date: Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E332E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: TEM: polymerization question 10, 23 -- thread-index: AcaWJKO/b+LSDKGdS4WFWVdZe1OTcA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 22 Jun 2006 17:52:34.0055 (UTC) FILETIME=[A3F7C170:01C69624] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MHqYXe002752 ==============================End of - Headers==============================
==============================Original Headers============================== 21, 30 -- From Geoffrey_Williams-at-brown.edu Thu Jun 22 13:25:35 2006 21, 30 -- Received: from pyxis.services.brown.edu (pyxis.services.brown.edu [128.148.106.154]) 21, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MIPYo7013369 21, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 13:25:35 -0500 21, 30 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 21, 30 -- by pyxis.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k5MIPWEj017229 21, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 14:25:32 -0400 (EDT) 21, 30 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 21, 30 -- Thu, 22 Jun 2006 14:25:32 -0400 21, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 21, 30 -- Content-class: urn:content-classes:message 21, 30 -- MIME-Version: 1.0 21, 30 -- Content-Type: text/plain; 21, 30 -- charset="us-ascii" 21, 30 -- Subject: RE: [Microscopy] TEM: polymerization question 21, 30 -- Date: Thu, 22 Jun 2006 14:25:30 -0400 21, 30 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F045EF925-at-MAIL1.AD.Brown.Edu} 21, 30 -- In-Reply-To: {200606221756.k5MHupsb009594-at-ns.microscopy.com} 21, 30 -- X-MS-Has-Attach: 21, 30 -- X-MS-TNEF-Correlator: 21, 30 -- Thread-Topic: [Microscopy] TEM: polymerization question 21, 30 -- Thread-Index: AcaWJT7Ysv/EcAo/RIO1sAvS0l72bwAA5ptQ 21, 30 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 21, 30 -- To: {Microscopy-at-microscopy.com} 21, 30 -- X-OriginalArrivalTime: 22 Jun 2006 18:25:32.0099 (UTC) FILETIME=[3EF95130:01C69629] 21, 30 -- X-Brown-Proofpoint: Not Infected 21, 30 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0606200000 definitions=3.0.0-0606220008 21, 30 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0606200000 definitions=3.0.0-0606220008 21, 30 -- Content-Transfer-Encoding: 8bit 21, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MIPYo7013369 ==============================End of - Headers==============================
few words to your question: from my experience this can happen, especially if you haven't "heated" and "ventilated" your oven after such "polymerizing" waste-/solvent-diluted resins....as this is with traces of water too, after e.g. staining in the same oven.... Do you use a ventilated oven (ventilation hole at the upper part of cabinet ? Another source could be moisture from room environment, if you are using the oven in a hood....
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We have been polymerizing our excess resins from infiltrations, etc., in the same oven we use for polymerizing our actual samples in resin. My question is whether this could have an effect on the quality of the resin blocks, maybe due to the solvents in the leftovers?
We had a couple batches of semi-gummy bears recently and this question has been resinating around the lab ever since. No samples lost, but a definite annoyance.
Thanks for any thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jun 22 12:52:34 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MHqYXe002752 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 12:52:34 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: TEM: polymerization question 10, 23 -- Date: Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E332E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: TEM: polymerization question 10, 23 -- thread-index: AcaWJKO/b+LSDKGdS4WFWVdZe1OTcA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 22 Jun 2006 17:52:34.0055 (UTC) FILETIME=[A3F7C170:01C69624] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MHqYXe002752 ==============================End of - Headers==============================
==============================Original Headers============================== 21, 28 -- From W.Muss-at-salk.at Thu Jun 22 13:36:43 2006 21, 28 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 21, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MIag6d023519 21, 28 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 13:36:42 -0500 21, 28 -- Received: from localhost (localhost [127.0.0.1]) 21, 28 -- by hermes.lks.at (Postfix) with ESMTP id AB4125A9044; 21, 28 -- Thu, 22 Jun 2006 20:36:40 +0200 (CEST) 21, 28 -- Received: from hermes.lks.at ([127.0.0.1]) 21, 28 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 21, 28 -- with ESMTP id 43922-06; Thu, 22 Jun 2006 20:36:40 +0200 (CEST) 21, 28 -- Received: from c1pa003 (unknown [192.168.42.3]) 21, 28 -- by hermes.lks.at (Postfix) with SMTP id 1F5FF5A9049; 21, 28 -- Thu, 22 Jun 2006 20:36:40 +0200 (CEST) 21, 28 -- Received: by localhost with Microsoft MAPI; Thu, 22 Jun 2006 20:36:39 +0200 21, 28 -- Message-ID: {01C6963B.9004AC20.W.Muss-at-salk.at} 21, 28 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 21, 28 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 21, 28 -- To: "'TindallR-at-missouri.edu'" {TindallR-at-missouri.edu} , 21, 28 -- "'microscopy-at-microscopy.com'" {microscopy-at-microscopy.com} 21, 28 -- Subject: AW: [Microscopy] TEM: polymerization question 21, 28 -- Date: Thu, 22 Jun 2006 20:36:37 +0200 21, 28 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 21, 28 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 21, 28 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 21, 28 -- MIME-Version: 1.0 21, 28 -- Content-Type: text/plain; charset="us-ascii" 21, 28 -- Content-Transfer-Encoding: 7bit 21, 28 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
I have a Reichert Ultracut (the model before the E) that has finally broken down. I'm looking for the following parts; motor (old part #7017-11097) and a microswitch (old part #SH 11101). I already know that the Leica folks back in Austria don't have anymore of these parts. I don't have the money to buy a new ultramicrotome, but if anyone out there is looking to sell a used one please contact me privately. I would definitely consider Reichert Ultracut models; LKB IV, V or Nova; RMC MT-7. Thanks.
Tom Bargar Core Electron Microscopy Research Facility UNMC 986395 Nebraska Medical Center Omaha, NE 68191-6395 tbargar-at-unmc.edu 402-559-7347
==============================Original Headers============================== 4, 17 -- From tbargar-at-unmc.edu Thu Jun 22 14:47:44 2006 4, 17 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MJliHX002538 4, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Jun 2006 14:47:44 -0500 4, 17 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 4, 17 -- by Outbound.unmc.edu (Proprietary) with ESMTP id D9F25A00DC 4, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Jun 2006 15:13:42 -0500 (CDT) 4, 17 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 4, 17 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 63DD539803F 4, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Jun 2006 15:13:41 -0500 (CDT) 4, 17 -- Subject: Looking for ultramicrotome parts or a used ultramicrotome 4, 17 -- To: Microscopy-at-MSA.Microscopy.Com 4, 17 -- Message-ID: {OF84E084F7.964DA68F-ON86257195.006AAD3E-86257195.006CBC8B-at-unmc.edu} 4, 17 -- From: Tom W Bargar {tbargar-at-unmc.edu} 4, 17 -- Date: Thu, 22 Jun 2006 14:47:41 -0500 4, 17 -- MIME-Version: 1.0 4, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
We are investigating the possibility of improving temperature stability in our JEM-2010F laboratory by using water-cooled radiant cooling panels. Thus far, we have found only one manufacturer: Energie Solaire in Switzerland. Can anyone recommend other manufacturers? Or is Energie Solaire the only game in town?
_________________________________
Joseph Kulik Research Associate Materials Research Institute The Pennsylvania State University 196 MRI Bldg University Park, PA 16802
I have used the methanol fixation for plants and dried using the Bal-Tec CPD030 and have had beautiful results. No problems with the seals.
owenha-at-csd.uwm.edu wrote:
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-- Karen L. Kelley ICBR Electron Microscopy Manager University of Florida ICBR Electron Microscopy Core Lab Building 747, Room 214 Box 118525 Gainesville Florida Lab: 352-392-1184 fax: 352-846-0251 Email: klk-at-biotech.ufl.edu Southeastern Microscopy Society Treasurer http://www.biotech.ufl.edu/EM/
==============================Original Headers============================== 7, 23 -- From klk-at-biotech.ufl.edu Thu Jun 22 17:49:12 2006 7, 23 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MMnCsS002893 7, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 17:49:12 -0500 7, 23 -- Received: from [128.227.60.41] (empc41719.dhcp.clas.ufl.edu [128.227.60.41]) 7, 23 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k5MMnAO15636178 7, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 18:49:10 -0400 7, 23 -- Message-ID: {449B1E66.5050403-at-biotech.ufl.edu} 7, 23 -- Date: Thu, 22 Jun 2006 18:49:10 -0400 7, 23 -- From: Karen Kelley {klk-at-biotech.ufl.edu} 7, 23 -- User-Agent: Mozilla Thunderbird 1.0.6 (Windows/20050716) 7, 23 -- X-Accept-Language: en-us, en 7, 23 -- MIME-Version: 1.0 7, 23 -- To: microscopy-at-microscopy.com 7, 23 -- Subject: Re: [Microscopy] Balzers Critical Point Drier solvents 7, 23 -- References: {200606221636.k5MGaZKd027432-at-ns.microscopy.com} 7, 23 -- In-Reply-To: {200606221636.k5MGaZKd027432-at-ns.microscopy.com} 7, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 23 -- Content-Transfer-Encoding: 7bit 7, 23 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 7, 23 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 7, 23 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 7, 23 -- 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 walter.bobrowski-at-pfizer.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: walter.bobrowski-at-pfizer.com Name: Walt Bobrowski
Question: In my attempts to apply IHC to epoxy embedded nerve tissue, the suggested pretreatment destroys the sections on the glass slides. The paper is "Immunohistochemical staining of epoxy resin sections of nerve tissue". Appl Immunohistochem Mol Morphol 2005;13:292-294". Pretreatment is 50% potassium ethoxide, rehydration, 10%H2O2, heated AR. The problem seems to be the combination of ethoxide and heated AR, as both are required, yet there is no section adherence problem with one OR the other. All advice regaring IHC of resin sections on glass slides much appreciated!
Best regards, Walt Bobrowski Pfizer Global R&D Ann Arbor, MI
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both aribaudo-at-triprinceton.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: aribaudo-at-triprinceton.org Name: Anthony Ribaudo
Organization: TRI Princeton
Title-Subject: [Filtered] Computer lockup problem on Zeiss 910 TEM
Question:
The computer screen on the Zeiss TEM I was using tends to get locked up when you hit the return button which is one the buttons in two rows one above and one below the computer scren. The return button is depressed to return to the master computer screen after viewing specific screens, for vacuum, camera, etc. to check/change operating parameters. The lockup affects other console controls as well such as the button one uses to toggle between coarse and fine magnification. Is it possible to unlock the computer without shutting the system down?
Anthony Ribaudo TRI Princeton Princeton, NJ 08542 aribaudo-at-triprinceton.org 609-430-4835
I have a friend who needs some 2"x 2" glass slides, 1 - 2 mm thick; does anyone have a source? -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 1, 16 -- From schooley-at-mcn.org Thu Jun 22 18:30:46 2006 1, 16 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 1, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MNUkLc001112 1, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Jun 2006 18:30:46 -0500 1, 16 -- Received: from [66.52.139.204] (helo=[10.0.1.2]) 1, 16 -- by dns4.mcn.org with esmtpa (Exim 4.60) 1, 16 -- (envelope-from {schooley-at-mcn.org} ) 1, 16 -- id J1ABZ6-0008MU-4S; Thu, 22 Jun 2006 16:30:43 -0700 1, 16 -- Mime-Version: 1.0 1, 16 -- Message-Id: {a06200700c0c0d5dd8fb0-at-[10.0.1.2]} 1, 16 -- Date: Thu, 22 Jun 2006 16:22:24 -0700 1, 16 -- To: Microscopy-at-MSA.Microscopy.Com 1, 16 -- From: Caroline Schooley {schooley-at-mcn.org} 1, 16 -- Subject: LM : 2 x 2 glass slides 1, 16 -- Cc: pswab-at-unitysemi.com 1, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Owen P. Mills wrote: ================================================== Does anyone know where I can get a bit of solid lutetium oxide appropriate for a microprobe standard? ================================================== See URL http://www.2spi.com/catalog/standards/aweb/lutetium.shtml
It is available commercially from SPI Supplies.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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==============================Original Headers============================== 14, 30 -- From cgarber-at-2spi.com Thu Jun 22 18:40:10 2006 14, 30 -- Received: from s-utl01-lopop.stsn.net (s-utl01-lopop.stsn.net [217.118.122.13]) 14, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5MNe986011239 14, 30 -- for {microscopy-at-msa.microscopy.com} ; Thu, 22 Jun 2006 18:40:10 -0500 14, 30 -- Received: from s-utl01-lopop.stsn.net ([127.0.0.1]) 14, 30 -- by s-utl01-lopop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006062300394923168 14, 30 -- for {microscopy-at-msa.microscopy.com} ; Fri, 23 Jun 2006 00:39:49 +0100 14, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 14, 30 -- tests=ALL_TRUSTED: -2.867,AWL: 0.178,BAYES_00: -1.665, 14, 30 -- SARE_RECV_ADDR: 0.027 14, 30 -- X-Spam-Level: 14, 30 -- Received: from ibm1x23g2abfyg ([10.112.7.242]) 14, 30 -- by s-utl01-lopop.stsn.net 14, 30 -- for microscopy-at-msa.microscopy.com; 14, 30 -- Fri, 23 Jun 2006 00:39:46 +0100 14, 30 -- Message-ID: {032401c69655$23753bf0$f207700a-at-ibm1x23g2abfyg} 14, 30 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 14, 30 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 14, 30 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 14, 30 -- Subject: Lutetium oxide standard for microanalysis 14, 30 -- Date: Thu, 22 Jun 2006 19:39:32 -0400 14, 30 -- MIME-Version: 1.0 14, 30 -- Content-Type: text/plain; 14, 30 -- charset="Windows-1252" 14, 30 -- X-Priority: 3 14, 30 -- X-MSMail-Priority: Normal 14, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 14, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 14, 30 -- Content-Transfer-Encoding: 8bit 14, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MNe986011239 ==============================End of - Headers==============================
they are another Swiss Company, but with US reps. Documentation on their site is pretty good. Their rep came out to visit us and helped us workup a proposal based upon their components for upgrades of one of our older labs.
Having said this, I should also say that ANL is currently also considering a local specialty contractor to build custom cooling panels for our new SAMM Lab building. If you touch base with me in about a month or two I can let you know which we decided on and how it goes. One way or the other we will have radiant cooling panels in SAMM Lab.
Nestor Your Friendly Neighborhood SysOp
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==============================Original Headers============================== 13, 14 -- From zaluzec-at-aaem.amc.anl.gov Thu Jun 22 18:42:42 2006 13, 14 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 13, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MNgfSo015407 13, 14 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 18:42:42 -0500 13, 14 -- Mime-Version: 1.0 13, 14 -- X-Sender: zaluzec-at-microscopy.com (Unverified) 13, 14 -- Message-Id: {p06110405c0c0d6f0730f-at-[206.69.208.22]} 13, 14 -- In-Reply-To: {200606222125.k5MLPtFa014032-at-ns.microscopy.com} 13, 14 -- References: {200606222125.k5MLPtFa014032-at-ns.microscopy.com} 13, 14 -- Date: Thu, 22 Jun 2006 18:42:47 -0500 13, 14 -- To: microscopy-at-microscopy.com 13, 14 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 13, 14 -- Subject: Radiant Cooling Panels 13, 14 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
I have a question involving darkfield imaging. We are using a Philips CM-10 and the imaging seems to be fine with one important exception. We have been capturing the area on film in standard brightfield mode and the images come out fine. Then we switch to darkfield by just pressing the DF button. The captured DF image is often is strongly directionally blurred, as though there is a lot of movement during capture. When the image is again checked in brightfield it looks fine.
Is it possible that there is charging in DF mode that could cause this but not in BF mode? Any other thoughts?
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://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 6, 21 -- From dsherman-at-purdue.edu Thu Jun 22 20:47:08 2006 6, 21 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5N1l8mE000674 6, 21 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 20:47:08 -0500 6, 21 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Thu, 22 Jun 2006 21:47:07 -0400 6, 21 -- Received: from 74.132.211.81 ([74.132.211.81]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 6, 21 -- Fri, 23 Jun 2006 01:47:05 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.2.3.060209 6, 21 -- Date: Thu, 22 Jun 2006 21:47:09 -0400 6, 21 -- Subject: TEM Darkfield imaging 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C0C0C05D.387E%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: TEM Darkfield imaging 6, 21 -- Thread-Index: AcaWZvBbLsG1xgJaEduvUQAKlcoUxg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 23 Jun 2006 01:47:07.0798 (UTC) FILETIME=[EFA42360:01C69666] ==============================End of - Headers==============================
Rock people go for odd slide sizes [petrographic slides seem quite small though], try : http://www.lakeside-products.com/html/std_slides.html
for 457 Plain 2" x 3" x 1mm thick slides (not 2x2 though). I expect they are out there somewhere as it sounds like a good old Victorian standard size, and I'm sure they could be custom made otherwise.
This link is from: http://www.uh.edu/~rmaddock/IRGO/microslides.html
There are thick 2" glass plates available : http://wardsci.com/product.asp_Q_pn_E_IG0009278_A_Glass+Plates
I have used laser cut CR-39 clear plastic 1mm thick for larger sizes - but they were for neutron induced fission fragment auto-radiographs and quite expensive at Ł1 or so each.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: schooley-at-mcn.org [mailto:schooley-at-mcn.org] Sent: 23 June 2006 00:35 To: keith.morris-at-ucl.ac.uk
I have a friend who needs some 2"x 2" glass slides, 1 - 2 mm thick; does anyone have a source? -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 1, 16 -- From schooley-at-mcn.org Thu Jun 22 18:30:46 2006 1, 16 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 1, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MNUkLc001112 1, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Jun 2006 18:30:46 -0500 1, 16 -- Received: from [66.52.139.204] (helo=[10.0.1.2]) 1, 16 -- by dns4.mcn.org with esmtpa (Exim 4.60) 1, 16 -- (envelope-from {schooley-at-mcn.org} ) 1, 16 -- id J1ABZ6-0008MU-4S; Thu, 22 Jun 2006 16:30:43 -0700 1, 16 -- Mime-Version: 1.0 1, 16 -- Message-Id: {a06200700c0c0d5dd8fb0-at-[10.0.1.2]} 1, 16 -- Date: Thu, 22 Jun 2006 16:22:24 -0700 1, 16 -- To: Microscopy-at-MSA.Microscopy.Com 1, 16 -- From: Caroline Schooley {schooley-at-mcn.org} 1, 16 -- Subject: LM : 2 x 2 glass slides 1, 16 -- Cc: pswab-at-unitysemi.com 1, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 20, 27 -- From keith.morris-at-ucl.ac.uk Fri Jun 23 04:38:13 2006 20, 27 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 20, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5N9cDHn020726 20, 27 -- for {Microscopy-at-microscopy.com} ; Fri, 23 Jun 2006 04:38:13 -0500 20, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 20, 27 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 20, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 20, 27 -- id 1Fti78-0001ku-AG 20, 27 -- for Microscopy-at-microscopy.com; Fri, 23 Jun 2006 10:38:10 +0100 20, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 20, 27 -- To: {Microscopy-at-microscopy.com} 20, 27 -- Subject: RE: [Microscopy] LM : 2 x 2 glass slides 20, 27 -- Date: Fri, 23 Jun 2006 10:38:07 +0100 20, 27 -- Message-ID: {000601c696a8$bbcaa590$7b865290-at-keithhigrade} 20, 27 -- MIME-Version: 1.0 20, 27 -- Content-Type: text/plain; 20, 27 -- charset="iso-8859-1" 20, 27 -- X-Mailer: Microsoft Office Outlook 11 20, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 20, 27 -- Thread-Index: AcaWVI35N84p+/g+TK+2YgFbhUdTMgAUVytQ 20, 27 -- In-Reply-To: {200606222335.k5MNZOsP008321-at-ns.microscopy.com} 20, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 20, 27 -- X-UCL-MailScanner: Found to be clean 20, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 20, 27 -- X-Spam-Status: No 20, 27 -- Content-Transfer-Encoding: 8bit 20, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5N9cDHn020726 ==============================End of - Headers==============================
Sounds like you either don't have the DF tilt alignments done properly, or are running in tilted DarkField mode.
You should be tilting the illumination to bring the post specimen diffracted beam down the optic axis so that there are minimal aberrations as well as have the Objective aperture centered. This is generally called centered Dark Field Mode.
Try to get hold of Eric Stach at Purdue (Engineering College) and have him come over to your lab and have a look. He should be able to sort it out what is wrong reasonably quickly.
Nestor Your Friendly Neighborhood SysOp
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
===========================================
==============================Original Headers============================== 13, 16 -- From zaluzec-at-aaem.amc.anl.gov Fri Jun 23 10:31:00 2006 13, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 13, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5NFUxjT004783 13, 16 -- for {microscopy-at-microscopy.com} ; Fri, 23 Jun 2006 10:31:00 -0500 13, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 13, 16 -- by aaem.amc.anl.gov (8.12.11.20060308/8.12.10) with ESMTP id k5NFUtMS004117 13, 16 -- for {microscopy-at-microscopy.com} ; Fri, 23 Jun 2006 10:30:59 -0500 13, 16 -- Mime-Version: 1.0 13, 16 -- Message-Id: {p06110404c0c1b6812e07-at-[146.139.72.105]} 13, 16 -- In-Reply-To: {200606230147.k5N1l8wn000681-at-ns.microscopy.com} 13, 16 -- References: {200606230147.k5N1l8wn000681-at-ns.microscopy.com} 13, 16 -- Date: Fri, 23 Jun 2006 10:30:54 -0500 13, 16 -- To: microscopy-at-microscopy.com 13, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 13, 16 -- Subject: Re: [Microscopy] TEM Darkfield imaging 13, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
I used to get 50 x 75mm slides from VWR. Cat No. 48300-207 and 48 x 60 mm cover slips Cat No. 48404-142
David Rothbard BEP Washington, DC
keith.morris-at-ucl.ac.uk wrote:
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==============================Original Headers============================== 7, 20 -- From rothbardd-at-netscape.net Fri Jun 23 11:38:24 2006 7, 20 -- Received: from imo-d01.mx.aol.com (imo-d01.mx.aol.com [205.188.157.33]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5NGcMEe016441 7, 20 -- for {microscopy-at-microscopy.com} ; Fri, 23 Jun 2006 11:38:23 -0500 7, 20 -- Received: from rothbardd-at-netscape.net 7, 20 -- by imo-d01.mx.aol.com (mail_out_v38_r7.5.) id w.4ed.229faf (22683) 7, 20 -- for {microscopy-at-microscopy.com} ; Fri, 23 Jun 2006 12:38:17 -0400 (EDT) 7, 20 -- Received: from netscape.net (mow-m01.webmail.aol.com [64.12.184.129]) by air-in04.mx.aol.com (v109.13) with ESMTP id MAILININ44-589b449c18f991; Fri, 23 Jun 2006 12:38:17 -0400 7, 20 -- Date: Fri, 23 Jun 2006 12:38:17 -0400 7, 20 -- From: rothbardd-at-netscape.net 7, 20 -- To: microscopy-at-microscopy.com 7, 20 -- Subject: RE: LM : 2 x 2 glass slides 7, 20 -- MIME-Version: 1.0 7, 20 -- Message-ID: {4A16DD70.372CC5D1.034D9A6A-at-netscape.net} 7, 20 -- X-Mailer: Atlas Mailer 2.0 7, 20 -- X-AOL-IP: 199.196.144.13 7, 20 -- X-AOL-Language: english 7, 20 -- Content-Type: text/plain; charset=iso-8859-1 7, 20 -- Content-Transfer-Encoding: 8bit 7, 20 -- X-Spam-Flag: NO ==============================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 tttan-at-simtech.a-star.edu.sg as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: tttan-at-simtech.a-star.edu.sg Name: TT Tan
Organization: SIMTech
Title-Subject: [Filtered] Electron beam Divergence
Question: Hello to All,
I am trying to find information on electron beam divergence Vs E-field.
For a given distance between anode and cathode, will the angular divergence increase with increasing E-field?
In addition, I am interested to know the divergence of E-Beam produced by a SINGLE strand of carbon nanotube.
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Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Filtered] Postdoctoral Position Open
Question: POST-DOCTORAL POSITION in Transmission Electron Microscopy of Nanodimensional Materials at The Pennsylvania State University
A postdoctoral position is available in the area of transmission electron microscopy in support of the Penn State MRSEC on Nanoscale Science beginning August 1, 2006. The position will be highly collaborative, working with groups studying the growth, properties and device integration of superconducting nanowires and semiconducting nanowires and heterostructures. Additional knowledge of semiconductor/superconductor electronic properties is beneficial, but not essential. The ideal candidate for this position will have experience in HRTEM, STEM, EDS and EELS. Strong communication skills and the ability to work in an interdisciplinary team environment are essential. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
OK, first - I have no life. I'm in the lab on Saturday cleaning the SEM Wehnelt assembly (I don't have a spare). Scrubbing, scrubbing, scrubbing to get that tough tungsten oxide coating off the business end. My mind begins to wander, and I recall showing a student the day before how thin the gold coating is on SEM specimens by wiping it off the bell jar of the sputter coater with a swipe of a lens tissue. Wouldn't it be nice if the junk on the Wehnelt came off as easily? Hmm....
So, has anybody tried this, i.e., sputter coating the Wehnelt orifice, with, say, gold? Would it help, hurt, make no difference, or be disastrous? Maybe someone out there has already tried this, and lived to tell about it....
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
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Email: martimor-at-nmsu.edu Name: Marti
Organization: NMSU
Title-Subject: [Filtered] travel question
Question: Sorry this is not a technical question. I was wondering if anyone has traveled with their diamond knife in their carry-on baggage? Any hassles at the airport? I am a very concerned student who must transport one soon.
MM asked the following: ============================================================ Question: Sorry this is not a technical question. I was wondering if anyone has traveled with their diamond knife in their carry-on baggage? Any hassles at the airport? I am a very concerned student who must transport one soon. ============================================================= By and large, the people who are trained to look at some kind of an x-ray image of what is in a carry on bag are trained to take a further look at anything not looking familiar to them. I can't think of anything that would look less "familiar" than a diamond knife.
You will then have the dilemma of trying to explain what it is or leaving it behind along with the confiscated cigarette lighters, pocket knives, scissors, and other items of unknown or potential danger. You will also run the real risk of missing your flight.
We always advise that when transporting something that is not "dangerous goods" in checked luggage, since on an x-ray scan, it could look "unfamiliar", when the suitcase is opened, there will be a note stating, in big letters, "This is not dangerous" and then proceed to tell what it really is. A print out from a website that explains what it is would help. You want to explain that it is not dangerous but if opened, it would very well destroy its value as a research tool. Having this done on the letter head of your university would be of additional help. The message should be signed and when I carry something like this, and using SPI Supplies "letterhead" paper, I like to put in my social security number and FEI (e.g. the tax) number of the company. The bag invariably gets opened, but the item being carried has so far been left undisturbed.
But whatever you do, don't ever stretch the truth and don't ever attempt to carry dangerous goods in checked luggage or any other way on a commercial airplane. At the worst, if it is thought this was done intentionally, you could not only be fined a large amount of money but you could end up on the "no fly" list of that airline.
Now having said all of that, why don't you just send it via one of the overnight courier services to your destination point and avoid the risk that someone might get curious about what is a diamond knife....it might sound like something that could be made into a piece of jewelry.
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
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==============================Original Headers============================== 16, 30 -- From cgarber-at-2spi.com Sat Jun 24 13:31:42 2006 16, 30 -- Received: from s-utl01-lopop.stsn.net (s-utl01-lopop.stsn.net [217.118.122.13]) 16, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5OIVfkB002936 16, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, 24 Jun 2006 13:31:41 -0500 16, 30 -- Received: from s-utl01-lopop.stsn.net ([127.0.0.1]) 16, 30 -- by s-utl01-lopop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006062419312828180 16, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, 24 Jun 2006 19:31:28 +0100 16, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 16, 30 -- tests=ALL_TRUSTED: -2.867,AWL: 0.179,BAYES_00: -1.665, 16, 30 -- SARE_RECV_ADDR: 0.027 16, 30 -- X-Spam-Level: 16, 30 -- Received: from ibm1x23g2abfyg ([10.104.227.217]) 16, 30 -- by s-utl01-lopop.stsn.net 16, 30 -- for microscopy-at-msa.microscopy.com; 16, 30 -- Sat, 24 Jun 2006 19:31:25 +0100 16, 30 -- Message-ID: {01c501c697bc$64217d40$d9e3680a-at-ibm1x23g2abfyg} 16, 30 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 16, 30 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 16, 30 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 16, 30 -- Subject: Travelling with a diamond knife 16, 30 -- Date: Sat, 24 Jun 2006 14:31:11 -0400 16, 30 -- MIME-Version: 1.0 16, 30 -- Content-Type: text/plain; 16, 30 -- charset="Windows-1252" 16, 30 -- X-Priority: 3 16, 30 -- X-MSMail-Priority: Normal 16, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 16, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 16, 30 -- Content-Transfer-Encoding: 8bit 16, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5OIVfkB002936 ==============================End of - Headers==============================
Here is the July 2006 Microscopy Today table of contents. I will close the subscription list for this issue on Thursday June 29, 2006.
Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$50 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Ron Anderson, Editor ============================== Imaging New Paths for Malarial Parasites Stephen W. Carmichael and Jon E. Rosenblatt, Mayo Clinic
Examination of the Gospel of Judas Using An Integrated Approach to Ink Characterization J.G. Barabe, K.A. Martin, E.F. Schumacher, J.R. Swider, and A.S. Teetsov, McCrone Associates, Inc., Westmont, IL
Active Optics Improve Microscope’s Field of View B. Potsaid and J.T. Wen, Rensselaer Polytechnic Institute, Troy, NY
An Introduction to the Helium Ion Microscope J. Morgan, J. Notte, R. Hill, B. Ward, ALIS Corporation, Peabody, MA
Solutions to the Problem of Substitution of ERL 4221 for Vinyl Cyclohexene Dioxide in Spurr Low Viscosity Embedding Formulations E. Ann Ellis, Texas A&M University, College Station, TX
Atom Probe Tomography Defines Mainstream Microscopy at the Atomic Scale T.F. Kelly1, K. Thompson1, E.A. Marquis2, and D.J. Larson; 1 1Imago Scientific Instruments Corp., Madison, WI, 2 Sandia National Laboratory, Livermore, CA
Keeping Life in Focus New Systems Prevent Z-axis Drift in Time Lapse Studies Edward Lachica, Olympus America Inc., Center Valley, PA
Infrared Chemical Imaging: Semi-Quantitative Analysis of Components J. Tarr, K. Nishikida, F. Izzia, Thermo Electron Corporation
Creating Pseudocolor Images using ImageJ Joel B. Sheffield, Temple University, Philadelphia PA
Nanoparticle Visualization with an AFM Paul West, Natasha Starostina, Pacific Nanotechnology, Tustin, CA
The Identification of Particles in a Polymer Film Using Nano-Thermal Analysis David Grandy & Kevin Kjoller, Anasys Instruments Corp., Santa Barbara, CA
Environmental Contamination Sources and Control in High Resolution Scanning Electron Microscopy Ronald Vane and Vince Carlino, XEI Scientific, Redwood City, CA
Water — A Clean “Glue” to Attach Hydrophilic Plates to an AFM Sample Stage Yang Gan, University of Newcastle, Callaghan, NSW, Australia
Protect Your Detectors C. Michael Stanley, Chroma Technology, VT
Dissolving Osmium Tetroxide the Easy Way Debby Sherman, Purdue University, West Lafayette, IN
Industry News
NetNotes SAMPLE PREPARATION - Critical point drier valves SAMPLE PREPARATION – negative staining enterococci SAMPLE PREPARATION – dispersing powder samples SAMPLE PREPARATION - metallic glasses SAMPLE PREPARATION - Time course experiment SAMPLE PREPARATION – Formvar film problems SAMPLE PREPARATION – Solvents that do not affect Formvar SAMPLE PREPARATION - Gelatin as the embedding media SAMPLE PREPARATION - neutralizing glutaraldehyde MICROTOMY – Diamond knife damage MICROTOMY – JB-4 resin MICROTOMY – sections moving around MICROTOMY – Biofilms on latex LM – field of view LM - quantitative fluorescence TEM - lead citrate TEM - immunolocalization SEM - roots for EM SEM - imaging sample with magnetic substrate SEM - ultrafiltration membranes SEM – red blood cells EM - platinum nanoparticles LM – fluorescence in plastic resins TEM - viral particles MICROSCOPY – imaging carbon nanotubes MICROSCOPY – imaging protein crystals
Index of Advertisers
==============================Original Headers============================== 22, 18 -- From microscopytoday-at-tampabay.rr.com Sat Jun 24 14:15:41 2006 22, 18 -- Received: from ms-smtp-04.tampabay.rr.com (ms-smtp-04.tampabay.rr.com [65.32.5.134]) 22, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5OJFfZ3017226 22, 18 -- for {Microscopy-at-Microscopy.Com} ; Sat, 24 Jun 2006 14:15:41 -0500 22, 18 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 22, 18 -- by ms-smtp-04.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k5OJFXhO012979; 22, 18 -- Sat, 24 Jun 2006 15:15:35 -0400 (EDT) 22, 18 -- Message-ID: {449D8F53.4020200-at-tampabay.rr.com} 22, 18 -- Date: Sat, 24 Jun 2006 15:15:31 -0400 22, 18 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 22, 18 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 22, 18 -- MIME-Version: 1.0 22, 18 -- To: Listserver {Microscopy-at-Microscopy.Com} , 22, 18 -- Confocal Listserver {confocal-at-listserv.buffalo.edu} 22, 18 -- Subject: Microscopy Today July 2006 Table of Contents 22, 18 -- Content-Type: text/plain; charset=windows-1252; format=flowed 22, 18 -- Content-Transfer-Encoding: 8bit 22, 18 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
For a much quicker method for cleaning cathodes take a look at Hits and Tips on our web site?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {jehrman-at-mta.ca} To: {protrain-at-emcourses.com} Sent: Saturday, June 24, 2006 3:08 PM
G'day,
I have to agree with Chuck's comments about trvelling with scientific equipment. A colleage was travelling from East to West Germany with some gear. One of the things was an ion gun for a surface analysis instrument. Unfortunately it was found in his baggage and when asked what it was, said "An ion GUN". Some 6 hours later and after a fairly torrid chat with security guards, eventually a University staff member was called in to explain things.
It was the last time he went to East Germany, firstly because he didn't want to go back and secondly because they wouldn't let him back in.
So, not only do you have to be careful where you put them, you have to be careful what you call these things.
Cheers
Colin Veitch.
==============================Original Headers============================== 7, 30 -- From Colin.Veitch-at-csiro.au Sat Jun 24 17:30:10 2006 7, 30 -- Received: from act-MTAout3.csiro.au (act-MTAout3.csiro.au [150.229.7.39]) 7, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5OMU9W0007284 7, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, 24 Jun 2006 17:30:10 -0500 7, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=AZ0KntvNYdIliGQoXPTfDR1afwbuAjIwyvM/5YFiaGhlYHs96OAeQvme8/hWbrQzW+jrGr5dQaNRmfModWfCgYgRn0o+x0K+qiiJJr6z6qjR5TSWZaFMXlz03ew50JsR; 7, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 7, 30 -- by act-MTAout3.csiro.au with ESMTP; 25 Jun 2006 08:30:42 +1000 7, 30 -- X-IronPort-AV: i="4.06,172,1149429600"; 7, 30 -- d="scan'208"; a="99941147:sNHT22305256" 7, 30 -- Received: from exvicn2-mel.nexus.csiro.au ([138.194.3.62]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 7, 30 -- Sun, 25 Jun 2006 08:30:07 +1000 7, 30 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exvicn2-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 7, 30 -- Sun, 25 Jun 2006 08:30:07 +1000 7, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 7, 30 -- content-class: urn:content-classes:message 7, 30 -- MIME-Version: 1.0 7, 30 -- Content-Type: text/plain; 7, 30 -- charset="Windows-1252" 7, 30 -- Subject: Re Travelling with a diamond knife 7, 30 -- Date: Sun, 25 Jun 2006 08:30:05 +1000 7, 30 -- Message-ID: {32CDDDAA7161394599F0025494915749024883-at-exvic5-gex.nexus.csiro.au} 7, 30 -- X-MS-Has-Attach: 7, 30 -- X-MS-TNEF-Correlator: 7, 30 -- Thread-Topic: Re Travelling with a diamond knife 7, 30 -- Thread-Index: AcaX3byyLhufgC7GTnCgrur1UiadCg== 7, 30 -- From: {Colin.Veitch-at-csiro.au} 7, 30 -- To: {microscopy-at-msa.microscopy.com} 7, 30 -- X-OriginalArrivalTime: 24 Jun 2006 22:30:07.0724 (UTC) FILETIME=[BF277AC0:01C697DD] 7, 30 -- Content-Transfer-Encoding: 8bit 7, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5OMU9W0007284 ==============================End of - Headers==============================
I guess that today, we need alternate, travel correct names for our historically everyday stuff. I'll start with:
Ion gun=energy emitter Diamond knife=precision excising tool Wire cutters=metal atom boundary separating device/tool Field emission gun=energy emitter Whenelt gun assembly=energy emitter assembly
sigh.....
gary g.
At 03:32 PM 6/24/2006, you wrote:
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==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Sat Jun 24 17:59:42 2006 13, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5OMxfRs017725 13, 20 -- for {microscopy-at-microscopy.com} ; Sat, 24 Jun 2006 17:59:42 -0500 13, 20 -- Received: (qmail 29474 invoked from network); 24 Jun 2006 15:59:41 -0700 13, 20 -- Received: by simscan 1.1.0 ppid: 29471, pid: 29472, t: 0.1646s 13, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 20 -- by qsmtp3 with SMTP; 24 Jun 2006 15:59:41 -0700 13, 20 -- Message-Id: {7.0.1.0.2.20060624155439.02502df0-at-gaugler.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 20 -- Date: Sat, 24 Jun 2006 15:59:42 -0700 13, 20 -- To: Colin.Veitch-at-csiro.au 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] Re Travelling with a diamond knife 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200606242232.k5OMWdeW010452-at-ns.microscopy.com} 13, 20 -- References: {200606242232.k5OMWdeW010452-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
The gold coating likely came easily because it is thin. Plus, it is not homogeneous. If you look at gold sputter coatings they look like spider webs. For low mag work, this is fine. Plus, they will change depending on your coating equipment. hence, high rez FE work is done with Au/Pd, Ir, Pt or Pd at low vacuum.
The gold coating on apertures is most likely done by plating, plasma deposition, evaporation or some other method that puts down a really nice even thin film.
Therein lies your culprit and vulnerability. The thin film can be easily removed via mechanical means. So trying to clean it would more than likely remove it.
I think you are stuck with what you have. The blue plague from the W filament is tough to remove. Pol works but takes time.
gary g.
At 07:09 AM 6/24/2006, you wrote:
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==============================Original Headers============================== 11, 21 -- From gary-at-gaugler.com Sat Jun 24 19:37:10 2006 11, 21 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5P0bA1N029047 11, 21 -- for {microscopy-at-microscopy.com} ; Sat, 24 Jun 2006 19:37:10 -0500 11, 21 -- Received: (qmail 21433 invoked from network); 24 Jun 2006 17:37:10 -0700 11, 21 -- Received: by simscan 1.1.0 ppid: 21430, pid: 21431, t: 0.1642s 11, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 21 -- by qsmtp2 with SMTP; 24 Jun 2006 17:37:09 -0700 11, 21 -- Message-Id: {7.0.1.0.2.20060624173101.024b8a78-at-gaugler.com} 11, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 21 -- Date: Sat, 24 Jun 2006 17:37:10 -0700 11, 21 -- To: jehrman-at-mta.ca 11, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 21 -- Subject: Re: [Microscopy] What would happen if ...... sputter coating 11, 21 -- Wehnelt assembly 11, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 21 -- In-Reply-To: {200606241409.k5OE9pak004774-at-ns.microscopy.com} 11, 21 -- References: {200606241409.k5OE9pak004774-at-ns.microscopy.com} 11, 21 -- Mime-Version: 1.0 11, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Sonicate in diluted ammonia, sudsy ammonia if possible, and the tungsten reside will come off easily. I usually dilute 2 ammonia:1 water. This hint originally came from the listserve but I have forgotten who provided it.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
On 6/24/06 10:10 AM, "jehrman-at-mta.ca" {jehrman-at-mta.ca} wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } OK, first - I have no life. I'm in the lab on Saturday cleaning the SEM } Wehnelt assembly } (I don't have a spare). Scrubbing, scrubbing, scrubbing to get that } tough tungsten oxide } coating off the business end. My mind begins to wander, and I recall } showing a student } the day before how thin the gold coating is on SEM specimens by wiping } it off the bell } jar of the sputter coater with a swipe of a lens tissue. Wouldn't it be } nice if the junk on the } Wehnelt came off as easily? Hmm.... } } So, has anybody tried this, i.e., sputter coating the Wehnelt orifice, } with, say, gold? Would it } help, hurt, make no difference, or be disastrous? Maybe someone out } there has already } tried this, and lived to tell about it.... } } Cheers, } } Jim }
==============================Original Headers============================== 8, 24 -- From dsherman-at-purdue.edu Sat Jun 24 21:53:17 2006 8, 24 -- Received: from 1061exfe03.adpc.purdue.edu (1061exfe03.adpc.purdue.edu [128.210.63.225]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5P2rHx9008449 8, 24 -- for {microscopy-at-microscopy.com} ; Sat, 24 Jun 2006 21:53:17 -0500 8, 24 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe03.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 8, 24 -- Sat, 24 Jun 2006 22:53:14 -0400 8, 24 -- Received: from 74.132.211.81 ([74.132.211.81]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 8, 24 -- Sun, 25 Jun 2006 02:53:12 +0000 8, 24 -- User-Agent: Microsoft-Entourage/11.2.3.060209 8, 24 -- Date: Sat, 24 Jun 2006 22:53:17 -0400 8, 24 -- Subject: Re: [Microscopy] What would happen if ...... sputter coating 8, 24 -- Wehnelt assembly 8, 24 -- From: Debby Sherman {dsherman-at-purdue.edu} 8, 24 -- To: {jehrman-at-mta.ca} , "message to: MSA list" {microscopy-at-microscopy.com} 8, 24 -- Message-ID: {C0C372DD.395C%dsherman-at-purdue.edu} 8, 24 -- Thread-Topic: [Microscopy] What would happen if ...... sputter coating 8, 24 -- Wehnelt assembly 8, 24 -- Thread-Index: AcaYAoJLwK87iAP1EduvUQAKlcoUxg== 8, 24 -- In-Reply-To: {200606241410.k5OEApUK005707-at-ns.microscopy.com} 8, 24 -- Mime-version: 1.0 8, 24 -- Content-type: text/plain; 8, 24 -- charset="US-ASCII" 8, 24 -- Content-transfer-encoding: 7bit 8, 24 -- X-OriginalArrivalTime: 25 Jun 2006 02:53:14.0651 (UTC) FILETIME=[80E55EB0:01C69802] ==============================End of - Headers==============================
On Sat, 24 Jun 2006 16:34:10 -0600, {Colin.Veitch-at-csiro.au} wrote:
} So, not only do you have to be careful where you put them, you have to } be careful what you call these things.
I realize that this may not be possible for many colleagues on limited budgets, but I no longer travel with any baggage _at all_. I FedEx my baggage ahead to the desk staff at my hotel, and travel with just my laptop- and FedEx the stuff home on the return as well, with a preprinted airbill. Needless to say, I no longer travel at all if I can help it: but if you have to, there's no reason to have to deal with the Neanderthals that have the power of go and no-go at airports. And this is especially true if there might be irreplaceable and/or unexplainable hardware (or analytical samples!) along for the ride. The random anonymous box gets there just fine, and is waiting for you at the desk on your arrival.
If you don't have the budget for this, I'm sad for you: but for me, the time spent in explaining anything covers the cost pretty well. I once spent a day and a half in Montreal's Dorval airport trying to explain some spare parts for a Leitz micrometer stage that looked wrong to a bag-screener... No more. Think about the time wasted when justifying this expenditure to The Powers That Be.
Standard disclaimer applies: I have no connection to FedEx other than as a satisfied and grateful customer, yadda yadda...
-- Scott Griffith ISES-LLC 9745 Steeplechase Drive Franktown, CO 80116 303-660-2541 voice 303-660-2542 fax skod-at-ises-llc.com
==============================Original Headers============================== 6, 19 -- From skod-at-ises-llc.com Sun Jun 25 00:26:26 2006 6, 19 -- Received: from trenco2.ises-llc.com (ises-llc.com [72.19.169.107]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5P5QQn7020305 6, 19 -- for {microscopy-at-msa.microscopy.com} ; Sun, 25 Jun 2006 00:26:26 -0500 6, 19 -- Received: from localhost (thrale.ises-llc.com [192.168.168.10]) 6, 19 -- by trenco2.ises-llc.com (8.12.10/8.12.10) with ESMTP id k5P54Fqd013799 6, 19 -- for {microscopy-at-msa.microscopy.com} ; Sat, 24 Jun 2006 23:04:15 -0600 (MDT) 6, 19 -- To: microscopy-at-msa.microscopy.com 6, 19 -- Subject: Re: [Microscopy] Re Travelling with a diamond knife 6, 19 -- References: {200606242234.k5OMYAnB012971-at-ns.microscopy.com} 6, 19 -- Message-ID: {op.tbor3dicgdwb11-at-localhost} 6, 19 -- Date: Sat, 24 Jun 2006 23:27:03 -0600 6, 19 -- From: "Scott Griffith" {skod-at-ises-llc.com} 6, 19 -- Organization: ISES-LLC 6, 19 -- Content-Type: text/plain; format=flowed; delsp=yes; charset=iso-8859-1 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Transfer-Encoding: 8bit 6, 19 -- In-Reply-To: {200606242234.k5OMYAnB012971-at-ns.microscopy.com} 6, 19 -- User-Agent: Opera M2/8.54 (MacPPC, build 2200) ==============================End of - Headers==============================
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Question: Hi, I want to know is there any technique available to increase the resolution of afm or fesem image of a polymer porous membrane (I want to have pics on nano sacle i.e. 50 to 1 nm) by impregnating a self settling gel/flouroscent resin in micropores of the porous membrane. or any other technique available.
"kuldeep" wrote: ===================================================== I want to know is there any technique available to increase the resolution of afm or fesem image of a polymer porous membrane (I want to have pics on nano sacle i.e. 50 to 1 nm) by impregnating a self settling gel/flouroscent resin in micropores of the porous membrane. or any other technique available. ====================================================== Did you mean 0.50 to 1 nm?
The limitation on the resolution in a FESEM image of a polymer, other than the instrument performance, usually is either insufficient contrast or too thick of a (conductive) coating thickness. I would suggest you consider the OPC osmium plasma coater, see URL http://www.2spi.com/catalog/osmi-coat.html The metal coating becomes conductive at slightly less than 1 nm, a much thinner thickness than any sputtered coating. You see a lot more when the metal coating is 1 nm instead of 10 or more nm in thickness.
We would be happy to do a demo coating for you. Contact me off-line if you would like to set up such a demo.
Disclaimer: SPI Supplies distributes the OPC Osmium Plasma Coaters and would have a vested interest in seeing more being purchased.
Chuck
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==============================Original Headers============================== 18, 29 -- From cgarber-at-2spi.com Sun Jun 25 15:51:12 2006 18, 29 -- Received: from s-utl01-lopop.stsn.net (s-utl01-lopop.stsn.net [217.118.122.13]) 18, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5PKpBbK004025 18, 29 -- for {microscopy-at-msa.microscopy.com} ; Sun, 25 Jun 2006 15:51:11 -0500 18, 29 -- Received: from s-utl01-lopop.stsn.net ([127.0.0.1]) 18, 29 -- by s-utl01-lopop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006062521510530041 18, 29 -- ; Sun, 25 Jun 2006 21:51:05 +0100 18, 29 -- X-Spam-Status: No, hits=0.0 required=9.9 18, 29 -- tests=ALL_TRUSTED: -2.867,AWL: -0.383,BAYES_00: -1.665, 18, 29 -- SARE_RECV_ADDR: 0.027,SARE_SUB_BETTER: 1.113 18, 29 -- X-Spam-Level: 18, 29 -- Received: from ibm1x23g2abfyg ([10.104.227.20]) 18, 29 -- by s-utl01-lopop.stsn.net; 18, 29 -- Sun, 25 Jun 2006 21:51:04 +0100 18, 29 -- Message-ID: {005f01c69899$1120d360$14e3680a-at-ibm1x23g2abfyg} 18, 29 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 18, 29 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 18, 29 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 18, 29 -- Subject: Better resolution on a FESEM specimen 18, 29 -- Date: Sun, 25 Jun 2006 16:50:51 -0400 18, 29 -- MIME-Version: 1.0 18, 29 -- Content-Type: text/plain; 18, 29 -- charset="Windows-1252" 18, 29 -- X-Priority: 3 18, 29 -- X-MSMail-Priority: Normal 18, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 18, 29 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 18, 29 -- Content-Transfer-Encoding: 8bit 18, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5PKpBbK004025 ==============================End of - Headers==============================
Does anyone have a recommendation for software to generate simulated electron diffraction patterns. I am familiar with Desktop Microscopist but cannot find a source for the software. I need to be able to generate simultaneous diffraction from the matrix as well as 1-4 second phases (with particular orientation relationships) and account for double diffraction as well. Any help would be greatly appreciated.
Many thanks in advance, Jim
==============================Original Headers============================== 3, 28 -- From mcnaney1-at-llnl.gov Sun Jun 25 19:51:39 2006 3, 28 -- Received: from smtp-3.llnl.gov (smtp-3.llnl.gov [128.115.41.83]) 3, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5Q0pdIe017810 3, 28 -- for {microscopy-at-microscopy.com} ; Sun, 25 Jun 2006 19:51:39 -0500 3, 28 -- Received: from nspws-1.llnl.gov (localhost [127.0.0.1]) 3, 28 -- by smtp-3.llnl.gov (8.13.6/8.12.3/LLNL evision: 1.16 $) with ESMTP id k5Q0pbFk008883; 3, 28 -- Sun, 25 Jun 2006 17:51:37 -0700 (PDT) 3, 28 -- Received: (from oracle-at-localhost) 3, 28 -- by nspws-1.llnl.gov (8.12.3p2-20030917/8.12.3/Submit) id k5Q0pb6H020861; 3, 28 -- Sun, 25 Jun 2006 17:51:37 -0700 (PDT) 3, 28 -- X-Authentication-Warning: nspws-1.llnl.gov: oracle set sender to mcnaney1-at-llnl.gov using -f 3, 28 -- Received: from vpna-user-128-15-244-26.llnl.gov 3, 28 -- (vpna-user-128-15-244-26.llnl.gov [128.15.244.26]) by 3, 28 -- www-openlabnet.llnl.gov (Horde MIME library) with HTTP; Sun, 25 Jun 2006 3, 28 -- 17:51:37 -0700 3, 28 -- Message-ID: {20060625175137.kcr49kzo80k4ok4o-at-www-openlabnet.llnl.gov} 3, 28 -- Date: Sun, 25 Jun 2006 17:51:37 -0700 3, 28 -- From: mcnaney1-at-llnl.gov 3, 28 -- To: microscopy-at-microscopy.com 3, 28 -- Subject: simulated electron diffraction patterns 3, 28 -- MIME-Version: 1.0 3, 28 -- Content-Type: text/plain; 3, 28 -- charset=ISO-8859-1; 3, 28 -- DelSp="Yes"; 3, 28 -- format="flowed" 3, 28 -- Content-Disposition: inline 3, 28 -- Content-Transfer-Encoding: 7bit 3, 28 -- User-Agent: Internet Messaging Program (IMP) H3 (4.1) ==============================End of - Headers==============================
In the past I have suspended fine particles in methanol when preparing TEM grids of powders. This seem to work well with the crushed minerals, ceramics, etc that I generally examine. However, questions have been raised about possible problems with particulate emissions from coal fired power stations. In particular:
1) does methanol evaporation cause particle agglomeration on the grid? 2) does methanol use cause morphological changes of the particles? 3) are particles size-fractionated on the TEM grid during the evaporation process? 4) does methanol dissolve some phases?
My question to the Listserver is - should I avoid methanol and if so which solvents should I use and why?
I would appreciate your advice and particularly if references can be provided. Cheers,
Mark Blackford Treasurer, AMMS Inc Institute of Materials Engineering Science PMB 1 MENAI, NSW, Australia 2234 phone 61 2 9717 3027 fax 61 2 9543 7179 email mgb-at-ansto.gov.au
==============================Original Headers============================== 10, 27 -- From mgb-at-ansto.gov.au Sun Jun 25 21:08:56 2006 10, 27 -- Received: from tachyon.gw.ansto.gov.au (tachyon.gw.ansto.gov.au [137.157.8.253]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5Q28sST028650 10, 27 -- for {Microscopy-at-microscopy.com} ; Sun, 25 Jun 2006 21:08:55 -0500 10, 27 -- Received: (from uucp-at-localhost) 10, 27 -- by tachyon.gw.ansto.gov.au (8.11.7p1+Sun/8.11.7) id k5Q28o802383 10, 27 -- for {Microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 12:08:50 +1000 (EST) 10, 27 -- Received: from jenner.ansto.gov.au(137.157.59.25) by tachyon.gw.ansto.gov.au via csmap (V6.0) 10, 27 -- id srcAAA__aWPe; Mon, 26 Jun 06 12:08:50 +1000 10, 27 -- Received: from hadron.ansto.gov.au (hadron.ansto.gov.au [137.157.13.219]) 10, 27 -- by jenner.ansto.gov.au (8.12.10/8.12.10) with ESMTP id k5Q28rra017507 10, 27 -- for {Microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 12:08:53 +1000 10, 27 -- Received: from paradise.ansto.gov.au (paradise.ansto.gov.au [137.157.58.208]) 10, 27 -- by hadron.ansto.gov.au (8.9.3/8.9.3) with ESMTP id MAA06812 10, 27 -- for {Microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 12:08:51 +1000 (EST) 10, 27 -- Received: from [137.157.95.57] (m0500m.ansto.gov.au [137.157.95.57]) by paradise.ansto.gov.au with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2657.72) 10, 27 -- id MZ72HH40; Mon, 26 Jun 2006 12:08:52 +1000 10, 27 -- Mime-Version: 1.0 (Apple Message framework v750) 10, 27 -- Content-Transfer-Encoding: 7bit 10, 27 -- Message-Id: {FF352706-42D5-490F-9570-2FDFE542E977-at-ansto.gov.au} 10, 27 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 10, 27 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 10, 27 -- From: Mark Blackford {mgb-at-ansto.gov.au} 10, 27 -- Subject: TEM powder sample preparation 10, 27 -- Date: Mon, 26 Jun 2006 12:03:09 +1000 10, 27 -- X-Mailer: Apple Mail (2.750) 10, 27 -- X-ANSTO-MailScanner: Found to be clean ==============================End of - Headers==============================
A JEOL engineer once told me an amusing story of one of their guys years ago who disappeared for three days after he arrived in the USSR to install an "electron gun assembly". This is why they are now shipped as "anode chambers", apparently.
} } I guess that today, we need alternate, travel correct } names for our historically everyday stuff. I'll start with: } } Ion gun=energy emitter } Diamond knife=precision excising tool } Wire cutters=metal atom boundary separating device/tool } Field emission gun=energy emitter } Whenelt gun assembly=energy emitter assembly } } } sigh..... } } } } gary g. } } } At 03:32 PM 6/24/2006, you wrote: }
==============================Original Headers============================== 3, 27 -- From ard-at-ansto.gov.au Mon Jun 26 02:05:03 2006 3, 27 -- Received: from tachyon.gw.ansto.gov.au (anstogw.gw.ansto.gov.au [137.157.8.253]) 3, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5Q752JI011310 3, 27 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 02:05:03 -0500 3, 27 -- Received: (from uucp-at-localhost) 3, 27 -- by tachyon.gw.ansto.gov.au (8.11.7p1+Sun/8.11.7) id k5Q74wT09012; 3, 27 -- Mon, 26 Jun 2006 17:04:58 +1000 (EST) 3, 27 -- Received: from jenner.ansto.gov.au(137.157.59.25) by tachyon.gw.ansto.gov.au via csmap (V6.0) 3, 27 -- id srcAAAm5ayMr; Mon, 26 Jun 06 17:04:58 +1000 3, 27 -- Received: from hadron.ansto.gov.au (hadron.ansto.gov.au [137.157.13.219]) 3, 27 -- by jenner.ansto.gov.au (8.12.10/8.12.10) with ESMTP id k5Q74xra019733; 3, 27 -- Mon, 26 Jun 2006 17:04:59 +1000 3, 27 -- Received: from [137.157.95.82] (arthur.amat.ansto.gov.au [137.157.95.82]) 3, 27 -- by hadron.ansto.gov.au (8.9.3/8.9.3) with ESMTP id RAA25005; 3, 27 -- Mon, 26 Jun 2006 17:04:56 +1000 (EST) 3, 27 -- Mime-Version: 1.0 3, 27 -- X-Sender: ard-at-hadron.ansto.gov.au 3, 27 -- Message-Id: {v04210101c0c53472513d-at-[137.157.95.82]} 3, 27 -- In-Reply-To: {200606242300.k5ON0gKT019745-at-ns.microscopy.com} 3, 27 -- References: {200606242300.k5ON0gKT019745-at-ns.microscopy.com} 3, 27 -- Date: Mon, 26 Jun 2006 17:04:57 +1000 3, 27 -- To: gary-at-gaugler.com 3, 27 -- From: Arthur Day {ard-at-ansto.gov.au} 3, 27 -- Subject: Re: Travelling with a diamond knife 3, 27 -- Cc: microscopy-at-microscopy.com 3, 27 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 27 -- X-ANSTO-MailScanner: Found to be clean ==============================End of - Headers==============================
Just for the curious among you (which means all ;-)). Here are the results of my "weird idea" of reading fluorescence on an axiovert microscope on semi-thin sectionned samples (100-300 nm) embedded in Epon:
DAPI staining (UV light) is strongly quenched. However, Alexa488 works pretty well. The bad news is that I can only see the fluorescence, I cannot see my cells by phase contrast! Now I need to find a way to see the cells with this method. Next step is embedding in Araldite. Unfortunately we are not equipped to embed in Lowicryl.
Stéphane
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==============================Original Headers============================== 5, 18 -- From nizets2-at-yahoo.com Mon Jun 26 03:04:12 2006 5, 18 -- Received: from web37401.mail.mud.yahoo.com (web37401.mail.mud.yahoo.com [209.191.87.54]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5Q84Alc022149 5, 18 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 03:04:11 -0500 5, 18 -- Received: (qmail 58712 invoked by uid 60001); 26 Jun 2006 08:04:06 -0000 5, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 18 -- s=s1024; d=yahoo.com; 5, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 5, 18 -- b=A+0MwIFQK9Kn9nYV31RUnrQ1V2F4e1lNuzS4mJn8AZ0rQVor8gGcYY7GUCUpJNAAR1hUXb0Ps+HU0fHKzTbtG9+8AB/tUDKagFBBIsAyChrwxOnXMBZ3WyzKjnR0DDkEtkoMqjlRdjsvmS2fJaaxtN3fzMgfp+0zBh8nFHVxVew= ; 5, 18 -- Message-ID: {20060626080406.58710.qmail-at-web37401.mail.mud.yahoo.com} 5, 18 -- Received: from [80.122.101.102] by web37401.mail.mud.yahoo.com via HTTP; Mon, 26 Jun 2006 01:04:06 PDT 5, 18 -- Date: Mon, 26 Jun 2006 01:04:06 -0700 (PDT) 5, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 5, 18 -- Subject: Fluorescence in Epoxy resin 5, 18 -- To: microscopy-at-microscopy.com 5, 18 -- MIME-Version: 1.0 5, 18 -- Content-Type: text/plain; charset=iso-8859-1 5, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Judging by the number of responses over the weekend, it looks like I may not be the only one without a life..... ;-)
Over the years I have tried most of the cleaning techniques suggested. Generally, though, I've found that any soaking technique doesn't get rid of the hardest, baked on tungsten that seems to be the culprit for additional whisker growth. I've also used a Dremel tool off and on - that does help but I find I fiddle with the chuck and such almost as much as just doing it by hand. And more often than not, the rechargeable battery on mine is dead when I need it!
I did want to clarify one thing - I was dreaming of gold coating with the hope that it *would* come off easily. I wasn't assuming that it would survive a polishing session. And yes, gold comes off of the glass chamber of the sputter coater very easily, but I can also pull it off the fairly roughly machined surfaces of the specimen stage by pressing on a piece of double stick tape and giving it a quick yank. Hence my curiosity as to whether this would serve as a barrier to tungsten adhesion. This behavior doesn't of course involve heating the gold coating to a very high temperature.
Since the topic of the gold evaporation came up - does anyone know the temperature of a typical Wehnelt in operation? I understand that the filament itself is hot enough to evaporate tungsten, but the orifice is almost certainly at a lower temperature, because this is where deposition occurs.
As usual, thanks for putting up with my (left field) questions.
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I used to have 7 diamond knives and did tons of TEM grade microtoming on coatings, silica filled rubber, oak flooring, polyureathanes, AR stacks, and other materials. Two were always in various stages of resharpening. Microstar always suggested to send them back for resharpening under another name to avoid losing them. So...
Diamond knife = carbon crystal. I never lost one in shipping.
When the purchasing agent saw the costs for resharpening, she told me she could get my diamond knives resharpened for less at a scissors sharpening vendor in Youngstown, Ohio. She was the best purchasing agent and could get deals you wouldn't believe. I showed her a diamond knife. Case closed.
TiO2, zirconia, mica, etc in paint basecoats trash diamond knives. Even softer materials (like all silica rubber tire cleats) can damage the brittle diamond edge by pressure flaking.
Paul
At 06:00 PM 6/24/06 -0500, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 25 -- From beaurega-at-westol.com Mon Jun 26 07:29:58 2006 7, 25 -- Received: from smtp-gateway-6.winbeam.com (smtp-gateway-6.winbeam.com [64.84.97.71]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5QCTuPk015185 7, 25 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 07:29:57 -0500 7, 25 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 7, 25 -- by smtp-gateway-6.winbeam.com (8.13.1/8.12.8) with SMTP id k5QCToiQ000413 7, 25 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 08:29:50 -0400 7, 25 -- Received: (qmail 29292 invoked by uid 89); 26 Jun 2006 12:29:45 -0000 7, 25 -- Received: from pitts-69-72-12-247.dynamic-dialup.coretel.net (HELO millenium) (69.72.12.247) 7, 25 -- by mail.winbeam.com with SMTP; 26 Jun 2006 12:29:45 -0000 7, 25 -- Message-Id: {3.0.6.32.20060626083057.007f4890-at-pop3.norton.antivirus} 7, 25 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 7, 25 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 7, 25 -- Date: Mon, 26 Jun 2006 08:30:57 -0400 7, 25 -- To: microscopy-at-microscopy.com 7, 25 -- From: Beaurega {beaurega-at-westol.com} 7, 25 -- Subject: Re: [Microscopy] Travelling with a diamond knife 7, 25 -- In-Reply-To: {200606242300.k5ON089R018588-at-ns.microscopy.com} 7, 25 -- Mime-Version: 1.0 7, 25 -- Content-Type: text/plain; charset="us-ascii" 7, 25 -- X-Winbeam-MailScanner-Information: - Please contact Technical Support for more information 7, 25 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) 7, 25 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, 7, 25 -- score=0.154, required 6, autolearn=disabled, TW_TR 0.08, TW_XG 0.08) 7, 25 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
And whatever you do, NEVER attempt to carryon a Fully Slotted, Helically Coiled, Fiber-Intrusive Fastening Device.*
*Disclaimer: I did not make this one up. Thank the folks at the Pentagon and the vendor who received, I think, upwards of twenty bucks for each one of these they magnanimously supplied our armed forces in the service of preserving liberty.
gary g. wrote.....
I guess that today, we need alternate, travel correct names for our historically everyday stuff. I'll start with:
Ion gun=energy emitter Diamond knife=precision excising tool Wire cutters=metal atom boundary separating device/tool Field emission gun=energy emitter Whenelt gun assembly=energy emitter assembly
sigh.....
------------------------------------------------------------------- Pluralitas non ponenda est sine necessitate. (=Keep it simple, stupid) - William of Ockham
==============================Original Headers============================== 14, 21 -- From pgrover-at-bilbo.bio.purdue.edu Mon Jun 26 07:37:43 2006 14, 21 -- Received: from mailhub130.itcs.purdue.edu (mailhub130.itcs.purdue.edu [128.210.5.130]) 14, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5QCbhcN025574 14, 21 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 07:37:43 -0500 14, 21 -- Received: from paklabpgrover (dhcp155-201.bio.purdue.edu [128.210.155.201]) 14, 21 -- by mailhub130.itcs.purdue.edu (8.13.7/8.13.7/internal-smtp) with ESMTP id k5QCbggp016825 14, 21 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 08:37:42 -0400 14, 21 -- From: "pgrover" {pgrover-at-bilbo.bio.purdue.edu} 14, 21 -- To: {microscopy-at-microscopy.com} 14, 21 -- Subject: traveling with a wood screw 14, 21 -- Date: Mon, 26 Jun 2006 08:37:48 -0400 14, 21 -- Message-ID: {001601c6991d$5512baf0$c99bd280-at-paklabpgrover} 14, 21 -- MIME-Version: 1.0 14, 21 -- Content-Type: text/plain; 14, 21 -- charset="us-ascii" 14, 21 -- Content-Transfer-Encoding: 7bit 14, 21 -- X-Mailer: Microsoft Office Outlook 11 14, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 14, 21 -- Thread-Index: AcaZHVTSARc30hgxTTKOgwMEJEDCQg== 14, 21 -- X-PMX-Version: 5.1.2.240295 14, 21 -- X-PerlMx-Virus-Scanned: Yes ==============================End of - Headers==============================
Don't use word "knife" if asked what that part is. Call it "specimen prep tool", and you will be fine. Place label "Fragile, handle with care" on the packaging. Security screeners don't open fragile items, unless item looks suspicious on X-ray scan. A diamond knife does not. Worst that will happen - they will double check for explosives residue and X-ray package one more time separately from the rest of your carry-on. Takes few extra minutes.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: {martimor-at-nmsu.edu} To: {vitalylazar-at-att.net} Sent: Saturday, June 24, 2006 1:35 PM
Here's the scoop/question. We have a 100x that has ceased springing, seemingly locked down in a manner that is not conducive to quality imaging.
Short of sending it in to the main company for service ($$$$$$) are there any suggestions for getting the objective functional again?
It is one of the 100x 1.3 phase units from the big Z.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
I have worked with coal and coal-derived particulates although I have not worked with TEM preparations.
I would take those questions as standard ones that should be asked of any liquid used for dispersion of any material. I don't think methanol deserves any special attention with regard to coal ash but you should always be on the lookout for these possible interactions.
I would not expect any morphological changes in the particles unless you were dissolving a phase. I expect the minerals, oxides, and any glasses formed from them during combustion would be inert to methanol and most solvents. I think the carbon would be charred and also rather inert.
Agglomeration would be a surface chemistry question and is beyond me, but I would not expect any special problems with methanol.
I would be on the lookout for size segregation with most solvents depending on many factors. I suppose it would be worse for slower evaporation processes, but a lot probably depends on an individual's technique as much as the choice of liquid.
Bottom line: I would probably try methanol and if you verify that there is no obvious agglomeration or segregation, then call it good and don't argue with success.
Warren Straszheim Materials Analysis and Research Laboratory Iowa State University
________________________________________ Sent: Sun 6/25/2006 9:10 PM
HI Geoff, I've had limited luck unsticking oil immersion lenses once they are fully gummed up. In the early stages of gumminess, I've had some luck with repeatedly wiping around the lens, at the point where it telescopes into itself, with a Kimwipe or lens paper saturated with lens cleaning solution (Sparkle or Windex glass cleaner diluted with an equal part water). I wipe around the lens, wiggle it as best I can to telescope a bit, pull it out, wipe, etc, I have almost always had to give in and send the lens back to Zeiss to be rebuilt. Its costly, but they price it just low enough for the price to be a "bargain" compared to purchasing a new lens. Its an occupational hazard of being an oil immersion lens on an inverted microscope. In 8 years, I've had to send back 2 63x oil, and 1 25x multi-immersion lenses. The last time the 63 on my confocal got gummed, I replaced it with a new one, since Zeiss re-engineered those lenses and they have 3 times the working distance of the old ones.
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============================== 2, 24 -- From lcgould-at-med.cornell.edu Mon Jun 26 10:53:03 2006 2, 24 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 2, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5QFr28N004582 2, 24 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 10:53:03 -0500 2, 24 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 2, 24 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5QFqwQ2004187 2, 24 -- for {microscopy-at-microscopy.com} ; Mon, 26 Jun 2006 11:52:59 -0400 (EDT) 2, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 2, 24 -- by mpx1.med.cornell.edu 2, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 2, 24 -- with ESMTPA id {0J1H005HE5G7PB90-at-mpx1.med.cornell.edu} for 2, 24 -- microscopy-at-microscopy.com; Mon, 26 Jun 2006 11:52:58 -0400 (EDT) 2, 24 -- Date: Mon, 26 Jun 2006 11:46:27 -0400 2, 24 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 2, 24 -- Subject: Re: [Microscopy] Repair of 100x oil objective? 2, 24 -- In-reply-to: {200606261524.k5QFOcr7017791-at-ns.microscopy.com} 2, 24 -- Sender: lcgould-at-med.cornell.edu 2, 24 -- To: Geoffrey_Williams-at-brown.edu, 2, 24 -- Microscopy Listserver {microscopy-at-microscopy.com} 2, 24 -- Message-id: {p06230907c0c5afe6d41b-at-[140.251.48.23]} 2, 24 -- MIME-version: 1.0 2, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 2, 24 -- References: {200606261524.k5QFOcr7017791-at-ns.microscopy.com} 2, 24 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.6.26.83932 ==============================End of - Headers==============================
First, you wouldn't want to sputter the gold you would want to evaporate it. Sputter films are usually more adherent than evaporated films. Secondly, you are putting a material that is easily evaporated into a situation where it could be evaporated onto the wrong components, namely the insulator of the filament. In the gun, you typically have a very good pressure and high temperature and you are hitting the filament side of the wehnelt with electrons which would heat it up further.
Keep scrubbing the W off or go to a LaB6 which seems to come off a little easier. I also recommend plasma cleaning the wehnelt after solvent cleaning.
-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: jehrman-at-mta.ca [mailto:jehrman-at-mta.ca] Sent: Saturday, June 24, 2006 7:14 AM To: Walck-at-SouthBayTech.com
OK, first - I have no life. I'm in the lab on Saturday cleaning the SEM Wehnelt assembly (I don't have a spare). Scrubbing, scrubbing, scrubbing to get that tough tungsten oxide coating off the business end. My mind begins to wander, and I recall showing a student the day before how thin the gold coating is on SEM specimens by wiping it off the bell jar of the sputter coater with a swipe of a lens tissue. Wouldn't it be nice if the junk on the Wehnelt came off as easily? Hmm....
So, has anybody tried this, i.e., sputter coating the Wehnelt orifice, with, say, gold? Would it help, hurt, make no difference, or be disastrous? Maybe someone out there has already tried this, and lived to tell about it....
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I've never had a problem getting deposited tungsten off using a diluted ammonia solution in a sonicator (use in hood!). The only warning would be not to put the brass bits in the strong ammonia solution, microQ does a good enough job discoloring them :) although generally nothing a nice relaxing afternoon with some cotton cloths, cotton applicators and a tube of POL won't cure.
I would sonicate in the ammonia for a while (till the W was gone), then in warm water, then a polish with some POL, followed by some microQ sonication, hot water, and then 95% ETOH and blown dry with canned air. The Yellow metals generally start with the POL steps.
But as always, your experiences may vary... (in other words I see no need to put gold on the Wehnelt)
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: jehrman-at-mta.ca [mailto:jehrman-at-mta.ca] Sent: Monday, June 26, 2006 8:27 AM To: Williams, Geoffrey
Hi listers,
Judging by the number of responses over the weekend, it looks like I may not be the only one without a life..... ;-)
Over the years I have tried most of the cleaning techniques suggested. Generally, though, I've found that any soaking technique doesn't get rid of the hardest, baked on tungsten that seems to be the culprit for additional whisker growth. I've also used a Dremel tool off and on - that does help but I find I fiddle with the chuck and such almost as much as just doing it by hand. And more often than not, the rechargeable battery on mine is dead when I need it!
I did want to clarify one thing - I was dreaming of gold coating with the hope that it *would* come off easily. I wasn't assuming that it would survive a polishing session. And yes, gold comes off of the glass chamber of the sputter coater very easily, but I can also pull it off the fairly roughly machined surfaces of the specimen stage by pressing on a piece of double
stick tape and giving it a quick yank. Hence my curiosity as to whether this would serve as a barrier to tungsten adhesion. This behavior doesn't of course involve heating the gold coating to a very high temperature.
Since the topic of the gold evaporation came up - does anyone know the temperature of a typical Wehnelt in operation? I understand that the filament itself is hot enough to evaporate tungsten, but the orifice is almost certainly at a lower temperature, because this is where deposition occurs.
As usual, thanks for putting up with my (left field) questions.
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I've been digging through the book references and aside from a few mentions of CPD vs Freeze Dry in Postek there are nearly no mentions of vague comparisons for EtOH or EtOH followed by HMDS dehydration. Background: I am a hard core CPD fan. This procedure was put in place in the absence (prior to me arriving) of a CPD, and was used throughout the experiment. Now the shrinkage variable is called into question by a reviewer, specifically how much does it shrink using EtOH as a dehydration/drying agent?
Are there any references out there I may have overlooked? Or general rough gut feelings about shrinkage (in regards to sample drying following Karnovsky's primary and buffered Osmium tetroxide fixation)? The samples (as further background) are dorsal root ganglia cell cultures (from Rat).
Thank you in advance,
Geoff Williams Leduc Bioimaging Facility Manager Brown University
This Question was submitted to Ask-A-Microscopist by (fli-at-estee.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 26, 2006 at 14:08:23 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both fli-at-estee.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: fli-at-estee.com Name: Foo Wing Li
Organization: Estee Lauder Companies
Education: Undergraduate College
Location: Melville, NY
Title: Availability of cold-SEM stage
Question: I need access to a cold SEM stage in the NY metro area. I need it for a project in examining microstructures of a cream that we're studying.
We recently installed a refurbished hollow cone unit on our JEM 2010 LaB6, and I was hoping to pick people's brains on the listserv as far as performance limitations. If anyone has experience with HC units on a JEM 2010 scope, or any microscope for that matter, please email me off the server.
As an example of the type of questions I have, "When attempting to correct the tilt purity while the HC unit is on (i.e., make the beam a point instead of a ring), we find that we have to max out the tilt correction. Is this normal, or is there some way to correct this in the unit itself?".
Kind regards, Matt Olszta
Matthew Olszta, Ph.D. Postdoctoral Researcher Penn State University 194 Materials Research Institute Building University Park, PA 16802 Phone: (814) 863-1096 Fax: (814) 863-8561
==============================Original Headers============================== 5, 16 -- From mjo10-at-psu.edu Tue Jun 27 07:43:44 2006 5, 16 -- Received: from f04n01.cac.psu.edu (f04s01.cac.psu.edu [128.118.141.31]) 5, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5RChhb9023562 5, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 07:43:44 -0500 5, 16 -- Received: from dickeyeels.psu.edu (dickeyeels.mri.psu.edu [146.186.179.10] (may be forged)) 5, 16 -- by f04n01.cac.psu.edu (8.13.2/8.13.2) with ESMTP id k5RChbcg150474 5, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 08:43:42 -0400 5, 16 -- Message-Id: {7.0.0.16.2.20060627081101.024056a0-at-psu.edu} 5, 16 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.0.16 5, 16 -- Date: Tue, 27 Jun 2006 08:45:39 -0400 5, 16 -- To: Microscopy-at-microscopy.com 5, 16 -- From: Matt Olszta {mjo10-at-psu.edu} 5, 16 -- Subject: Hollow Cone JEM 2010 5, 16 -- Mime-Version: 1.0 5, 16 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 5, 16 -- X-Virus-Scanned: by amavisd-new ==============================End of - Headers==============================
This is probably the most stupid request in the history of this list, but I would need a detailed protocol to prepare poly-L-Lysin coverslips. Currently after incubating the coverslips with the polylysin solution we dry them under the hood and then autoclave them. Apparently autoclaving does not disturb the coating but the coverslips stick together and we cannot separate them without breaking them apart. I am sure it would be possible to wash them with ethanol to sterilize them but I don't know exactly how to do it. So in short I would need the very pratical details of how to sterilize the coverslips after coating to avoid them to stick together.
Stephane
P.S: my deepest apologizes for asking intellectually unchallenging questions :-D
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Tue Jun 27 09:59:54 2006 6, 18 -- Received: from web37411.mail.mud.yahoo.com (web37411.mail.mud.yahoo.com [209.191.87.64]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5RExrhP004145 6, 18 -- for {microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 09:59:53 -0500 6, 18 -- Received: (qmail 67907 invoked by uid 60001); 27 Jun 2006 14:59:53 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 18 -- b=AlxBqz/QBX9Bq8JnnnE4oyKwBn+AyF18b4k2lXpZG1HCF2et3o3Wr6uEJ1tsr0a9NIBWwqQXEh/mxbYYY+9KAMiuZP3TBhiiA4Ej2PGkJge19JDZBtSzEi4D0XlpTstcFSfP+3mVRVTXEKGG8bAZcJn1uYhkGPg3wwJoR094wYs= ; 6, 18 -- Message-ID: {20060627145953.67905.qmail-at-web37411.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37411.mail.mud.yahoo.com via HTTP; Tue, 27 Jun 2006 07:59:52 PDT 6, 18 -- Date: Tue, 27 Jun 2006 07:59:52 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: need a protocol to prepare polylysin-coated coverslips 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==============================
Take a large glass Petri dish and line it with a clean piece of filter paper. Place each coverslip on the filter paper so they don't overlap with each other. Insert into autoclave. Tedious but works well.
At 10:01 AM 06/27/06, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
ENGINEER SENIOR John M. Crowley Center for high Resolution Electron Microscopy (CHREM)
The John M. Crowley Center for High Resolution electron Microscopy (CHREM) at Arizona State University seeks an Engineer Senior to provide technical support of advanced electron microscopes and ion beam instruments. Deadline: 5pm, 7/24/06; if not filled, then every week thereafter until search is closed. Salary: $40,480 - $64,959/DOE. AA/EOE. For qualifications/application information, please see SR#O-124463 (Req. #0601123) at www.asu.edu/hr/jobs.
Please follow the instruction and make a proper contact.
Thanks.
Zhenquan Liu Senior Research Specialist Arizona State University PSA 213 Tempe, 85287-1704 Tel (480) 965 4512 Fax (480) 965 9004
==============================Original Headers============================== 3, 25 -- From Zhenquan.Liu-at-asu.edu Tue Jun 27 19:16:51 2006 3, 25 -- Received: from epo-int1.asu.edu (epo-int1.asu.edu [129.219.187.20]) 3, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5S0Gnj2001665 3, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 19:16:50 -0500 3, 25 -- Received: from EX03.asurite.ad.asu.edu (excl1-b0.asurite.ad.asu.edu [129.219.12.197]) 3, 25 -- by epo-int1.asu.edu (Switch-3.1.8/Switch-3.1.7/asu-postoffice-prod) with ESMTP id k5S0Gmt5018820; 3, 25 -- Tue, 27 Jun 2006 17:16:48 -0700 3, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 3, 25 -- Content-class: urn:content-classes:message 3, 25 -- MIME-Version: 1.0 3, 25 -- Content-Type: text/plain; 3, 25 -- charset="iso-8859-1" 3, 25 -- Subject: EM Job position at ASU 3, 25 -- Date: Tue, 27 Jun 2006 17:16:49 -0700 3, 25 -- Message-ID: {60D5D23B9F20DF4C8DC3991979F9094A78D38B-at-EX03.asurite.ad.asu.edu} 3, 25 -- X-MS-Has-Attach: 3, 25 -- X-MS-TNEF-Correlator: 3, 25 -- Thread-Topic: EM Job position at ASU 3, 25 -- Thread-Index: AcaaPXsomtvpU3cwQkK3fXaLT5E/bwACnV8g 3, 25 -- From: "Zhenquan Liu" {Zhenquan.Liu-at-asu.edu} 3, 25 -- To: {Microscopy-at-microscopy.com} 3, 25 -- Cc: {zhen1-at-asu.edu} 3, 25 -- X-Virus-Scanned: by amavisd-new 3, 25 -- Content-Transfer-Encoding: 8bit 3, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5S0Gnj2001665 ==============================End of - Headers==============================
ENGINEER SENIOR John M. Crowley Center for high Resolution Electron Microscopy (CHREM)
The John M. Crowley Center for High Resolution electron Microscopy (CHREM) at Arizona State University seeks an Engineer Senior to provide technical support of advanced electron microscopes and ion beam instruments. Deadline: 5pm, 7/24/06; if not filled, then every week thereafter until search is closed. Salary: $40,480 - $64,959/DOE. AA/EOE. For qualifications/application information, please see SR#O-124463 (Req. #0601123) at www.asu.edu/hr/jobs, 1) select Administrative and Staff, 2) Search Posting, 3) Req. # (just the #).
Please follow the instruction and make a proper contact.
Thanks.
Zhenquan Liu Senior Research Specialist Arizona State University PSA 213 Tempe, 85287-1704 Tel (480) 965 4512 Fax (480) 965 9004
==============================Original Headers============================== 8, 25 -- From Zhenquan.Liu-at-asu.edu Tue Jun 27 19:19:09 2006 8, 25 -- Received: from epo-int1.asu.edu (epo-int1.asu.edu [129.219.187.20]) 8, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5S0J6RG003879 8, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 19:19:08 -0500 8, 25 -- Received: from EX03.asurite.ad.asu.edu (excl1-b0.asurite.ad.asu.edu [129.219.12.197]) 8, 25 -- by epo-int1.asu.edu (Switch-3.1.8/Switch-3.1.7/asu-postoffice-prod) with ESMTP id k5S0J5GY020339; 8, 25 -- Tue, 27 Jun 2006 17:19:05 -0700 8, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 8, 25 -- Content-class: urn:content-classes:message 8, 25 -- MIME-Version: 1.0 8, 25 -- Content-Type: text/plain; 8, 25 -- charset="us-ascii" 8, 25 -- Subject: EM Job at ASU 8, 25 -- Date: Tue, 27 Jun 2006 17:19:06 -0700 8, 25 -- Message-ID: {60D5D23B9F20DF4C8DC3991979F9094A78D38C-at-EX03.asurite.ad.asu.edu} 8, 25 -- X-MS-Has-Attach: 8, 25 -- X-MS-TNEF-Correlator: 8, 25 -- Thread-Topic: EM Job at ASU 8, 25 -- Thread-Index: AcaaSG7SPB5kb3J/Tq+paCFxWMPeEg== 8, 25 -- From: "Zhenquan Liu" {Zhenquan.Liu-at-asu.edu} 8, 25 -- To: {Microscopy-at-microscopy.com} 8, 25 -- Cc: {zhen1-at-asu.edu} 8, 25 -- X-Virus-Scanned: by amavisd-new 8, 25 -- Content-Transfer-Encoding: 8bit 8, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5S0J6RG003879 ==============================End of - Headers==============================
Due a typing error in the previous posted e-mail for the name of Professor John Cowley, I am posting this job ad here again. I am very sorry about that. Zhenquan Liu
ENGINEER SENIOR John M. Cowley Center for High Resolution Electron Microscopy (CHREM)
The John M. Cowley Center for High Resolution Electron Microscopy (CHREM) at Arizona State University seeks an Engineer Senior to provide technical support of advanced electron microscopes and ion beam instruments. Deadline: 5pm, 7/24/06; if not filled, then every week thereafter until search is closed. Salary: $40,480 - $64,959/DOE. AA/EOE. For qualifications/application information, please see SR#O-124463 (Req. #0601123) at www.asu.edu/hr/jobs, 1) select Administrative and Staff, 2) Search Posting, 3) Req. # (just the #).
Please follow the instruction and make a proper contact.
Thanks.
Zhenquan Liu Senior Research Specialist Arizona State University PSA 213 Tempe, 85287-1704 Tel (480) 965 4512 Fax (480) 965 9004
==============================Original Headers============================== 8, 25 -- From Zhenquan.Liu-at-asu.edu Tue Jun 27 19:19:09 2006 8, 25 -- Received: from epo-int1.asu.edu (epo-int1.asu.edu [129.219.187.20]) 8, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5S0J6RG003879 8, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 19:19:08 -0500 8, 25 -- Received: from EX03.asurite.ad.asu.edu (excl1-b0.asurite.ad.asu.edu [129.219.12.197]) 8, 25 -- by epo-int1.asu.edu (Switch-3.1.8/Switch-3.1.7/asu-postoffice-prod) with ESMTP id k5S0J5GY020339; 8, 25 -- Tue, 27 Jun 2006 17:19:05 -0700 8, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 8, 25 -- Content-class: urn:content-classes:message 8, 25 -- MIME-Version: 1.0 8, 25 -- Content-Type: text/plain; 8, 25 -- charset="us-ascii" 8, 25 -- Subject: EM Job at ASU 8, 25 -- Date: Tue, 27 Jun 2006 17:19:06 -0700 8, 25 -- Message-ID: {60D5D23B9F20DF4C8DC3991979F9094A78D38C-at-EX03.asurite.ad.asu.edu} 8, 25 -- X-MS-Has-Attach: 8, 25 -- X-MS-TNEF-Correlator: 8, 25 -- Thread-Topic: EM Job at ASU 8, 25 -- Thread-Index: AcaaSG7SPB5kb3J/Tq+paCFxWMPeEg== 8, 25 -- From: "Zhenquan Liu" {Zhenquan.Liu-at-asu.edu} 8, 25 -- To: {Microscopy-at-microscopy.com} 8, 25 -- Cc: {zhen1-at-asu.edu} 8, 25 -- X-Virus-Scanned: by amavisd-new 8, 25 -- Content-Transfer-Encoding: 8bit 8, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5S0J6RG003879 ==============================End of - Headers==============================
==============================Original Headers============================== 13, 24 -- From Zhenquan.Liu-at-asu.edu Tue Jun 27 20:16:25 2006 13, 24 -- Received: from epo-int1.asu.edu (epo-int1.asu.edu [129.219.187.20]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5S1GPet022590 13, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 20:16:25 -0500 13, 24 -- Received: from EX03.asurite.ad.asu.edu (excl1-b0.asurite.ad.asu.edu [129.219.12.197]) 13, 24 -- by epo-int1.asu.edu (Switch-3.1.8/Switch-3.1.7/asu-postoffice-prod) with ESMTP id k5S1GNUN008630 13, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 18:16:23 -0700 13, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 24 -- Content-class: urn:content-classes:message 13, 24 -- MIME-Version: 1.0 13, 24 -- Content-Type: text/plain; 13, 24 -- charset="us-ascii" 13, 24 -- Subject: EM Job at ASU 13, 24 -- Date: Tue, 27 Jun 2006 18:16:24 -0700 13, 24 -- Message-ID: {60D5D23B9F20DF4C8DC3991979F9094A78D38E-at-EX03.asurite.ad.asu.edu} 13, 24 -- X-MS-Has-Attach: 13, 24 -- X-MS-TNEF-Correlator: 13, 24 -- Thread-Topic: EM Job at ASU 13, 24 -- Thread-Index: AcaaSSbDBqvrD9LfS7mQjokdvflrWwABnGYQ 13, 24 -- From: "Zhenquan Liu" {Zhenquan.Liu-at-asu.edu} 13, 24 -- To: {Microscopy-at-microscopy.com} 13, 24 -- X-Virus-Scanned: by amavisd-new 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5S1GPet022590 ==============================End of - Headers==============================
I would not bother with sending anything to Zeiss for repair. I had lousy service for my Zeiss LM objectives and stands but fixed this by dumping them for Olympus.
However, Zeiss SMT FESEMs are another issue. More poor service. It seems that there are just not enough people of talent to handle these sophisticated and delicate tools. This is probably a common problem amongst other brands. The tools sell but they do need occasional service.
gary g.
At 08:25 AM 6/26/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Tue Jun 27 20:58:13 2006 9, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5S1wDwI000731 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 20:58:13 -0500 9, 20 -- Received: (qmail 27383 invoked from network); 27 Jun 2006 18:58:12 -0700 9, 20 -- Received: by simscan 1.1.0 ppid: 27380, pid: 27381, t: 0.1671s 9, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 20 -- by qsmtp2 with SMTP; 27 Jun 2006 18:58:12 -0700 9, 20 -- Message-Id: {7.0.1.0.2.20060627185222.0250ecf8-at-gaugler.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 20 -- Date: Tue, 27 Jun 2006 18:58:14 -0700 9, 20 -- To: Geoffrey_Williams-at-brown.edu 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] Repair of 100x oil objective? 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200606261525.k5QFPkNb019647-at-ns.microscopy.com} 9, 20 -- References: {200606261525.k5QFPkNb019647-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
How about just the page that has the cam settings for the duration, thickness, and height controls? If I can get that, I can calibrate the wheels. There seem to be no service manuals floating around, from which photocopies can be made.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
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I posted this once before, but got no takers, so I thought I'd give it another go. I have manuals for the Philips EM300 and the RCA EMU-4. Anyone need them?
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Wed Jun 28 07:19:24 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5SCJO3d025925 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 07:19:24 -0500 3, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5SCrk4l013734 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 08:53:46 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Wed, 28 Jun 2006 08:19:21 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f06230907c0c823ef711c-at-[141.209.160.132]} 3, 20 -- Date: Wed, 28 Jun 2006 08:19:22 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: Philips and RCA manuals 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 28 Jun 2006 12:19:21.0582 (UTC) FILETIME=[1600F8E0:01C69AAD] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -4 () L_EXCH_MF 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Here is the link to the table of X-Ray energies from J. A. Bearden, in "X-Ray Wavelengths," Rev. Mod. Phys. 39, (1967) p.78.
http://xray.uu.se/hypertext/XREmission.html
Cheers, Henk
At 04:02 PM 06/28/06, opmills-at-mtu.edu wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Electron Microscopy Center Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a Mac !
===========================================
==============================Original Headers============================== 12, 16 -- From zaluzec-at-aaem.amc.anl.gov Wed Jun 28 16:21:48 2006 12, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 12, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5SLLlS5010004 12, 16 -- for {microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 16:21:47 -0500 12, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 12, 16 -- by aaem.amc.anl.gov (8.12.11.20060308/8.12.10) with ESMTP id k5SLLk9a003322 12, 16 -- for {microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 16:21:47 -0500 12, 16 -- Mime-Version: 1.0 12, 16 -- Message-Id: {p06110416c0c8a2da3e24-at-[146.139.72.105]} 12, 16 -- In-Reply-To: {200606281959.k5SJxdDW021219-at-ns.microscopy.com} 12, 16 -- References: {200606281959.k5SJxdDW021219-at-ns.microscopy.com} 12, 16 -- Date: Wed, 28 Jun 2006 16:21:45 -0500 12, 16 -- To: microscopy-at-microscopy.com 12, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 12, 16 -- Subject: Re: [Microscopy] need x-ray emission table 12, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
I am trying to do some nanoindentation experiments on a NiCoCrAlY coating which primarily contains two phases: NiAl and Ni-solid solution. For the experiments to be of any use I have to ensure that I remove the sub-surface damage layer created during mechanical polishing. I think electropolishing is a reasonable option provided I find the suitable electrolyte and conditions. So far perchloric/ acetic acid gives fairly good results but I was wondering whether there was a better solution that I could use? I am also open to other techniques for preparing damage free specimens.
Thanks,
Budhika Mendis
Postdoctoral fellow, Johns Hopkins University
==============================Original Headers============================== 5, 34 -- From mendis-at-poseidon.me.jhu.edu Wed Jun 28 16:44:29 2006 5, 34 -- Received: from ipex4.johnshopkins.edu (ipex4.johnshopkins.edu [128.220.2.151]) 5, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5SLiSrO020633 5, 34 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 16:44:28 -0500 5, 34 -- Received: from jhuml3.jhu.edu ([128.220.2.66]) 5, 34 -- by ipex4.johnshopkins.edu with ESMTP/TLS/DES-CBC3-SHA; 28 Jun 2006 17:44:22 -0400 5, 34 -- X-BrightmailFiltered: true 5, 34 -- X-Brightmail-Tracker: AAAAAA== 5, 34 -- X-IronPort-AV: i="4.06,189,1149480000"; 5, 34 -- d="scan'208"; a="162162817:sNHT22694776" 5, 34 -- Received: from poseidon.me.jhu.edu (poseidon.me.jhu.edu [128.220.2.27]) 5, 34 -- by jhuml3.jhu.edu (PMDF V6.2-X27 #31041) 5, 34 -- with ESMTP id {0J1L00JTMB1YAB-at-jhuml3.jhu.edu} for Microscopy-at-Microscopy.Com; 5, 34 -- Wed, 28 Jun 2006 17:44:22 -0400 (EDT) 5, 34 -- Received: from me.jhu.edu ([128.220.2.27]) 5, 34 -- by poseidon.me.jhu.edu (Sun Java System Messaging Server 6.1 HotFix 0.09 5, 34 -- (built Dec 14 2004)) with ESMTP id {0J1L0023QB1HDM10-at-poseidon.me.jhu.edu} for 5, 34 -- Microscopy-at-Microscopy.Com; Wed, 28 Jun 2006 17:44:05 -0400 (EDT) 5, 34 -- Received: from [128.220.28.72] by poseidon.me.jhu.edu (mshttpd); Wed, 5, 34 -- 28 Jun 2006 17:44:05 -0400 5, 34 -- Date: Wed, 28 Jun 2006 17:44:05 -0400 5, 34 -- From: Budhika Mendis {mendis-at-poseidon.me.jhu.edu} 5, 34 -- Subject: LM Electrolyte for superalloy coatings 5, 34 -- To: Microscopy-at-microscopy.com 5, 34 -- Message-id: {80581d816aa.44a2bfe5-at-me.jhu.edu} 5, 34 -- MIME-version: 1.0 5, 34 -- X-Mailer: Sun Java(tm) System Messenger Express 6.1 HotFix 0.09 (built Dec 14 5, 34 -- 2004) 5, 34 -- Content-type: text/plain; charset=us-ascii 5, 34 -- Content-language: en 5, 34 -- Content-transfer-encoding: 7BIT 5, 34 -- Content-disposition: inline 5, 34 -- X-Accept-Language: en 5, 34 -- Priority: normal ==============================End of - Headers==============================
Owen: one of my favorites is EM Periodic Table, a program authored by Scott Walck many years ago. It has a periodic chart layout and a table layout that can be translated to Excel. I got it at a Lehigh short course and I find it very handy. I'd be happy to share it with you. Scott calls this "beerware" meaning when we meet him at conferences, we owe him a beer if we use it.
Scott: you can claim your pitcher at M&M in Chicago. This program would be cheap at twice the price.
opmills-at-mtu.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Does anyone know where I can find a spreadsheet or text version of x- } ray emission lines? } } Thanks, } } OWen } } } } } Owen P. Mills } Director, Materials Characterization & Fabrication Facilities } Electron Optics Engineer, Applied Chemical & Morphological Analysis } Laboratory } } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills } } } } ==============================Original Headers============================== } 10, 31 -- From opmills-at-mtu.edu Wed Jun 28 14:59:39 2006 } 10, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) } 10, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5SJxdNZ021212 } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 14:59:39 -0500 } 10, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) } 10, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id k5SJxc9N010211 } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } 10, 31 -- Received: from node2.edge.dcsint.mtu.edu (node2.mtu.edu [141.219.69.2]) } 10, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k5SJxcCb016435 } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } 10, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) } 10, 31 -- by node2.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k5SJxbZH003257 } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } 10, 31 -- (envelope-from opmills-at-mtu.edu) } 10, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) } 10, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k5SJxbQP012706 } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:37 -0400 (EDT) } 10, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) } 10, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k5SJxbbF016425 } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:37 -0400 } 10, 31 -- Mime-Version: 1.0 (Apple Message framework v750) } 10, 31 -- Content-Transfer-Encoding: 7bit } 10, 31 -- Message-Id: {82FC92F2-DB6B-4300-BE2B-59C95CA02CBB-at-mtu.edu} } 10, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 10, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} } 10, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} } 10, 31 -- Subject: need x-ray emission table } 10, 31 -- Date: Wed, 28 Jun 2006 15:59:46 -0400 } 10, 31 -- X-Mailer: Apple Mail (2.750) } 10, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.6.28.123433 } 10, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' } ==============================End of - Headers============================== } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 5, 22 -- From r-holdford-at-ti.com Wed Jun 28 17:34:58 2006 5, 22 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5SMYwwA031196 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 28 Jun 2006 17:34:58 -0500 5, 22 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 5, 22 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k5SMYj7s012187; 5, 22 -- Wed, 28 Jun 2006 17:34:54 -0500 (CDT) 5, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 5, 22 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k5SMYi7g009682; 5, 22 -- Wed, 28 Jun 2006 17:34:45 -0500 (CDT) 5, 22 -- Message-ID: {44A30404.3050301-at-ti.com} 5, 22 -- Date: Wed, 28 Jun 2006 17:34:44 -0500 5, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 5, 22 -- Organization: SC Packaging Development -- FA Development 5, 22 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 5, 22 -- MIME-Version: 1.0 5, 22 -- To: opmills-at-mtu.edu, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- Subject: Re: [Microscopy] need x-ray emission table 5, 22 -- References: {200606282000.k5SK04LD021547-at-ns.microscopy.com} 5, 22 -- In-Reply-To: {200606282000.k5SK04LD021547-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Thanks for the comment about the program. I was suprised anyone was using it except for me. I use it to look up potential overlaps for X-ray and EELS peaks. I used several tables to try to get the best possible data available and cross-checked the values. For some lines that were not available, formulas were used to calculate them.
EMPeridodicTable is available on the EMMPDL. It is also incorporated in the program, EELS_Plot. Both are Visual Basic programs that you have to install n Windows. I believe that the table is in tab deliminated form or CSV form on one of the data files that are loaded into the pregram directory and that it can be brought into Excel without problems. You would have to extract the files and look at the data files or extract it out of the CAB file for the program.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
I have used a HC unit on a JEOL 2000FX. Actually I have used three units on three 2000FX's. It was critically important to get the microscope aligned properly. It was very critical to get the trans and tilt controls properly decoupled. It was also important not to scan it too fast. You could see a "glitch" in the pattern on the screen when it went too fast. If you recorded a slow scan, the "glitch" wasn't there.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-------- Original Message -------- } From: mjo10-at-psu.edu } Sent: Tuesday, June 27, 2006 8:49 AM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] Hollow Cone JEM 2010 } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } We recently installed a refurbished hollow cone unit on our JEM 2010 } LaB6, and I was hoping to pick people's brains on the listserv as far } as performance limitations. If anyone has experience with HC units } on a JEM 2010 scope, or any microscope for that matter, please email } me off the server. } } As an example of the type of questions I have, "When attempting to } correct the tilt purity while the HC unit is on (i.e., make the beam } a point instead of a ring), we find that we have to max out the tilt } correction. Is this normal, or is there some way to correct this in } the unit itself?". } } Kind regards, } Matt Olszta } } Matthew Olszta, Ph.D. } Postdoctoral Researcher } Penn State University } 194 Materials Research Institute Building } University Park, PA 16802 } Phone: (814) 863-1096 } Fax: (814) 863-8561 } } } ==============================Original Headers============================== } 5, 16 -- From mjo10-at-psu.edu Tue Jun 27 07:43:44 2006 } 5, 16 -- Received: from f04n01.cac.psu.edu (f04s01.cac.psu.edu [128.118.141.31]) } 5, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5RChhb9023562 } 5, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 07:43:44 -0500 } 5, 16 -- Received: from dickeyeels.psu.edu (dickeyeels.mri.psu.edu [146.186.179.10] (may be forged)) } 5, 16 -- by f04n01.cac.psu.edu (8.13.2/8.13.2) with ESMTP id k5RChbcg150474 } 5, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 27 Jun 2006 08:43:42 -0400 } 5, 16 -- Message-Id: {7.0.0.16.2.20060627081101.024056a0-at-psu.edu} } 5, 16 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.0.16 } 5, 16 -- Date: Tue, 27 Jun 2006 08:45:39 -0400 } 5, 16 -- To: Microscopy-at-microscopy.com } 5, 16 -- From: Matt Olszta {mjo10-at-psu.edu} } 5, 16 -- Subject: Hollow Cone JEM 2010 } 5, 16 -- Mime-Version: 1.0 } 5, 16 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } 5, 16 -- X-Virus-Scanned: by amavisd-new } ==============================End of - Headers==============================
==============================Original Headers============================== 5, 20 -- From walck-at-southbaytech.com Wed Jun 28 22:22:27 2006 5, 20 -- Received: from smtp02.safesecureweb.com (smtp02.safesecureweb.com [66.241.211.51]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5T3MQF0024513 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 22:22:27 -0500 5, 20 -- Received: from mail15.safesecureweb.com (ironhide.ad.safesecureweb.com [192.168.2.180]) 5, 20 -- by smtp02.safesecureweb.com (Spam Firewall) with ESMTP 5, 20 -- id 19AAC80A705; Wed, 28 Jun 2006 23:22:26 -0400 (EDT) 5, 20 -- MIME-Version: 1.0 5, 20 -- Date: Wed, 28 Jun 2006 23:22:07 -0400 5, 20 -- Content-Type: text/plain; 5, 20 -- charset=iso-8859-1 5, 20 -- Subject: re: [Microscopy] Hollow Cone JEM 2010 5, 20 -- From: Scott Walck {walck-at-southbaytech.com} 5, 20 -- Reply-To: Walck-at-southbaytech.com 5, 20 -- To: {mjo10-at-psu.edu} 5, 20 -- CC: {microscopy-at-microscopy.com} 5, 20 -- Message-ID: {6958a1212373455180a04dd4e3c24551-at-southbaytech.com} 5, 20 -- X-Virus-Scanned: by Barracuda Spam Firewall at safesecureweb.com 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5T3MQF0024513 ==============================End of - Headers==============================
The RCA EMU 4 and Philips EM300 manuals have been spoken for.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Thu Jun 29 10:43:59 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5TFhxti015245 3, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 29 Jun 2006 10:43:59 -0500 3, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5TGIJ4l032448 3, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 29 Jun 2006 12:18:19 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Thu, 29 Jun 2006 11:43:58 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f0623090ac0c9a560b1f2-at-[141.209.160.132]} 3, 20 -- Date: Thu, 29 Jun 2006 11:43:58 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: RCA and Philips manuals 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 29 Jun 2006 15:43:58.0395 (UTC) FILETIME=[D5F7B8B0:01C69B92] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -4 () L_EXCH_MF 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
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Email: mbsmith Name: Marianne B. Smith
Organization: Iowa State University
Title-Subject: [Filtered] Staining technique
Question: Hi, I have been asked to stain slides with Johansen's Safronin O and Fast Green FCF method, according to a recipe from Steve Ruzin's book. My problem:I cannot find a "well-known" supplier of Methyl cellosolve. Does anybody know where (from which company) to buy this chemical? Does it maybe have a different chemical name? I would appreciate your expertise and help. Marianne
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 mbsmith-at-iastate.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mbsmith-at-iastate.edu Name: Marianne B. Smith
Organization: ISU
Title-Subject: [Filtered] Staining technique
Question: Earlier today I asked a question about staining Safronin O and Fast Green FCF, with Johansen's method, out of the book of Ruzin. I cannot remember if I wrote down my complete e-mail address.I wanted to know where to buy Methyl cellosolve. Sorry, and thanks for any help and answer. Marianne
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Email: acruz04-at-cibnor.mx Name: Ariel Cruz
Organization: CIBNOR SC
Title-Subject: [Filtered] Problem with EDX
Question: I really need your help it but quick possible, notice that working with the EDX Inca Oxford 2000 suddenly accent of receiving sign and it marked me overflow, to what tries to restart but it sends me the windows that I also send you (overflow, automation error that it lacks a file MCPDLL.DLL). Deadtime to 99%.
Linear Formula: CH3OCH2CH2OH, Formula Weight: 76.09, CAS Number: 109-86-4
Notice: Be prepared: rather hazardous / toxic substance, cf.: http://dnr.wi.gov/org/aw/air/emission/nr438/pollutants/298.htm
EGME (2-Methoxyethanol; MethylCellosolve) Use information: This is a glycol ether. Coatings represent one of the important applications of gycolether solvents. Glycloethers are used in surface coating formulations, particularly in lacquers, enamels, water-borne coatings, phenolic varnishes, epoxy coatings, specific coatings for styrene and in non-grain-raising wood stains. They improve glosss and flow out in lacquers and the gloss and the gloss retention during baking in synthetic enamels. As a dye solvent in the textile, leather and printing industries, glycolethers increase the dye stability, aid the penetration of dyes and yields brighter colours. Glycolethers are also excellent solvents for insecticides, herbicides, grease and grim in speciality chemicals, nitrocellulose, acrylic, epoxy, polyamide and alkyd resin. As mutual solvent and coupler they are used in soluble oils, metal and glass cleaners, varnich removers and other speciality cleaners. Soap/hydrocarbon systems may be clarified by the addtions of glycolether because it is highley miscible with hydrocarbons, water and soaps. Because of its good thermal stability and excellent viscosity/temperature relationship glycolethers are particularly useful as diluent in hydraulic brake fluids. NR 445, NR407 Pollutant
Would it be possible to get - off list - some informations on the staining technique you have to use?
best wishes and good luck, Wolfgang Muss Salzburg, Austria
Zitat von mbsmith-at-ns.microscopy.com:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } 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 mbsmith as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: mbsmith } Name: Marianne B. Smith } } Organization: Iowa State University } } Title-Subject: [Filtered] Staining technique } } Question: Hi, } I have been asked to stain slides with Johansen's Safronin O and Fast Green } FCF method, according to a recipe from Steve Ruzin's book. My problem:I } cannot find a "well-known" supplier of Methyl cellosolve. Does anybody know } where (from which company) to buy this chemical? Does it maybe have a } different chemical name? I would appreciate your expertise and help. } Marianne } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Thu Jun 29 20:05:10 2006 } 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k5U159O4002826 } 6, 12 -- for {microscopy-at-microscopy.com} ; Thu, 29 Jun 2006 20:05:09 -0500 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110400c0ca29375b20-at-[206.69.208.22]} } 6, 12 -- Date: Thu, 29 Jun 2006 20:05:07 -0500 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: mbsmith-at-ns.microscopy.com (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: Staining technique } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
==============================Original Headers============================== 21, 40 -- From W.Muss-at-salk.at Fri Jun 30 01:01:25 2006 21, 40 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9]) 21, 40 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5U61Ou8006354 21, 40 -- for {microscopy-at-microscopy.com} ; Fri, 30 Jun 2006 01:01:24 -0500 21, 40 -- Received: from localhost (localhost [127.0.0.1]) 21, 40 -- by hermes.salk.at (Postfix) with ESMTP id 53BCCC3832; 21, 40 -- Fri, 30 Jun 2006 08:01:30 +0200 (CEST) 21, 40 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 21, 40 -- Received: from hermes.salk.at ([127.0.0.1]) 21, 40 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 21, 40 -- with ESMTP id GANKolqvN3eU; Fri, 30 Jun 2006 08:01:29 +0200 (CEST) 21, 40 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 21, 40 -- by hermes.salk.at (Postfix) with ESMTP id E4439C382C; 21, 40 -- Fri, 30 Jun 2006 08:01:29 +0200 (CEST) 21, 40 -- Received: from localhost (localhost [127.0.0.1]) 21, 40 -- by hermes.lks.at (Postfix) with ESMTP id D18F35A9026; 21, 40 -- Fri, 30 Jun 2006 08:01:29 +0200 (CEST) 21, 40 -- Received: from hermes.lks.at ([127.0.0.1]) 21, 40 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 21, 40 -- with ESMTP id 61668-03; Fri, 30 Jun 2006 08:01:29 +0200 (CEST) 21, 40 -- Received: from salk.at (webmail.salk.at [192.168.13.209]) 21, 40 -- by hermes.lks.at (Postfix) with ESMTP id 85E265A9020; 21, 40 -- Fri, 30 Jun 2006 08:01:29 +0200 (CEST) 21, 40 -- Received: from m160p027.adsl.highway.telekom.at (m160p027.adsl.highway.telekom.at [62.47.187.251]) 21, 40 -- by webmail.salk.at (IMP) with HTTP 21, 40 -- for {pawma-at-192.168.13.210} ; Fri, 30 Jun 2006 08:01:29 +0200 21, 40 -- Message-ID: {1151647289.44a4be39585db-at-webmail.salk.at} 21, 40 -- Date: Fri, 30 Jun 2006 08:01:29 +0200 21, 40 -- From: Wolfgang Muss {W.Muss-at-salk.at} 21, 40 -- To: mbsmith-at-ns.microscopy.com 21, 40 -- Cc: microscopy-at-microscopy.com, w.muss-at-salk.at 21, 40 -- Subject: [Microscopy]Re: Staining technique 21, 40 -- References: {200606300112.k5U1CrRX024277-at-ns.microscopy.com} 21, 40 -- In-Reply-To: {200606300112.k5U1CrRX024277-at-ns.microscopy.com} 21, 40 -- MIME-Version: 1.0 21, 40 -- Content-Type: text/plain; charset=ISO-8859-1 21, 40 -- Content-Transfer-Encoding: 8bit 21, 40 -- User-Agent: Internet Messaging Program (IMP) 3.2.8 21, 40 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 21, 40 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
----- Weitergeleitete Nachricht von Wolfgang Muss {W.Muss-at-salk.at} ----- Datum: Fri, 30 Jun 2006 08:01:29 +0200 Von: Wolfgang Muss {W.Muss-at-salk.at} Antwort an: Wolfgang Muss {W.Muss-at-salk.at} Betreff: [Microscopy]Re: Staining technique An: mbsmith-at-ns.microscopy.com
Linear Formula: CH3OCH2CH2OH, Formula Weight: 76.09, CAS Number: 109-86-4
Notice: Be prepared: rather hazardous / toxic substance, cf.: http://dnr.wi.gov/org/aw/air/emission/nr438/pollutants/298.htm
EGME (2-Methoxyethanol; MethylCellosolve) Use information: This is a glycol ether. Coatings represent one of the important applications of gycolether solvents. Glycloethers are used in surface coating formulations, particularly in lacquers, enamels, water-borne coatings, phenolic varnishes, epoxy coatings, specific coatings for styrene and in non-grain-raising wood stains. They improve glosss and flow out in lacquers and the gloss and the gloss retention during baking in synthetic enamels. As a dye solvent in the textile, leather and printing industries, glycolethers increase the dye stability, aid the penetration of dyes and yields brighter colours. Glycolethers are also excellent solvents for insecticides, herbicides, grease and grim in speciality chemicals, nitrocellulose, acrylic, epoxy, polyamide and alkyd resin. As mutual solvent and coupler they are used in soluble oils, metal and glass cleaners, varnich removers and other speciality cleaners. Soap/hydrocarbon systems may be clarified by the addtions of glycolether because it is highley miscible with hydrocarbons, water and soaps. Because of its good thermal stability and excellent viscosity/temperature relationship glycolethers are particularly useful as diluent in hydraulic brake fluids.
NB: NR 445, NR407 Pollutant
Would it be possible to get - off list - some informations on the staining technique you have to use?
best wishes and good luck, Wolfgang Muss Salzburg, Austria
Zitat von mbsmith-at-ns.microscopy.com: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } 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 mbsmith as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: mbsmith } Name: Marianne B. Smith } } Organization: Iowa State University } } Title-Subject: [Filtered] Staining technique } } Question: Hi, } I have been asked to stain slides with Johansen's Safronin O and Fast Green } FCF method, according to a recipe from Steve Ruzin's book. My problem:I } cannot find a "well-known" supplier of Methyl cellosolve. Does anybody know } where (from which company) to buy this chemical? Does it maybe have a } different chemical name? I would appreciate your expertise and help. } Marianne } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Thu Jun 29 20:05:10 2006 } 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k5U159O4002826 } 6, 12 -- for {microscopy-at-microscopy.com} ; Thu, 29 Jun 2006 20:05:09 -0500 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110400c0ca29375b20-at-[206.69.208.22]} } 6, 12 -- Date: Thu, 29 Jun 2006 20:05:07 -0500 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: mbsmith-at-ns.microscopy.com (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: Staining technique } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
----- Ende der weitergeleiteten Nachricht -----
----- Ende der weitergeleiteten Nachricht -----
==============================Original Headers============================== 29, 37 -- From W.Muss-at-salk.at Fri Jun 30 01:06:02 2006 29, 37 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9]) 29, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5U661gD009686 29, 37 -- for {microscopy-at-microscopy.com} ; Fri, 30 Jun 2006 01:06:01 -0500 29, 37 -- Received: from localhost (localhost [127.0.0.1]) 29, 37 -- by hermes.salk.at (Postfix) with ESMTP id E19DAC3870; 29, 37 -- Fri, 30 Jun 2006 08:06:17 +0200 (CEST) 29, 37 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 29, 37 -- Received: from hermes.salk.at ([127.0.0.1]) 29, 37 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 29, 37 -- with ESMTP id J9x10tPodziG; Fri, 30 Jun 2006 08:06:17 +0200 (CEST) 29, 37 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 29, 37 -- by hermes.salk.at (Postfix) with ESMTP id 82387C3864; 29, 37 -- Fri, 30 Jun 2006 08:06:17 +0200 (CEST) 29, 37 -- Received: from localhost (localhost [127.0.0.1]) 29, 37 -- by hermes.lks.at (Postfix) with ESMTP id 5B89A5A9046; 29, 37 -- Fri, 30 Jun 2006 08:06:17 +0200 (CEST) 29, 37 -- Received: from hermes.lks.at ([127.0.0.1]) 29, 37 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 29, 37 -- with ESMTP id 62083-09; Fri, 30 Jun 2006 08:06:17 +0200 (CEST) 29, 37 -- Received: from salk.at (webmail.salk.at [192.168.13.209]) 29, 37 -- by hermes.lks.at (Postfix) with ESMTP id 008DD5A9020; 29, 37 -- Fri, 30 Jun 2006 08:06:16 +0200 (CEST) 29, 37 -- Received: from m160p027.adsl.highway.telekom.at (m160p027.adsl.highway.telekom.at [62.47.187.251]) 29, 37 -- by webmail.salk.at (IMP) with HTTP 29, 37 -- for {pawma-at-192.168.13.210} ; Fri, 30 Jun 2006 08:06:16 +0200 29, 37 -- Message-ID: {1151647576.44a4bf58bd0a9-at-webmail.salk.at} 29, 37 -- Date: Fri, 30 Jun 2006 08:06:16 +0200 29, 37 -- From: Wolfgang Muss {W.Muss-at-salk.at} 29, 37 -- To: mbsmith-at-iastate.edu, microscopy-at-microscopy.com 29, 37 -- Subject: Fwd: Fwd: [Microscopy]Re: Staining technique 29, 37 -- MIME-Version: 1.0 29, 37 -- Content-Type: text/plain; charset=ISO-8859-1 29, 37 -- Content-Transfer-Encoding: 8bit 29, 37 -- User-Agent: Internet Messaging Program (IMP) 3.2.8 29, 37 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 29, 37 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
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Email: charlie-at-jtcorp.co.kr Name: Charlie Lee
Organization: JT Corp
Title-Subject: [Filtered] Schottky FE e-gun supplier for SEM
Question: I'm looking for a Schottky FE e-gun manufacturer for SEM. Is there anybody who knows suppliers ?
I appreciate information so much in advance.
Regards,
Charlie Lee JT Corp ---------------------------------------------------------------------------
==============================Original Headers============================== 8, 12 -- From zaluzec-at-microscopy.com Fri Jun 30 07:31:56 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 k5UCVsLx004384 8, 12 -- for {microscopy-at-microscopy.com} ; Fri, 30 Jun 2006 07:31:56 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: (Unverified) 8, 12 -- Message-Id: {p06110400c0caca1910e1-at-[206.69.208.22]} 8, 12 -- Date: Fri, 30 Jun 2006 07:31:53 -0500 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- From: charlie-at-jtcorp.co.kr (by way of MicroscopyListserver) 8, 12 -- Subject: viaWWW: Schottky FE e-gun supplier for SEM 8, 12 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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Title-Subject: [Filtered] RE : [Microscopy] viaWWW: Problem with EDX
Question: I had the same problem and solve it this way: On the back of the blue computer unit of the oxford system and at the bottom there is a fan in front of which stand a filter, take it out and clean it.
Put it back in and restart the machine, it should work out.
Diane
Diane Montpetit Centre de Recherche et de DČveloppement sur les Aliments 3600 Boul. Casavant Ouest, St-Hyacinthe, (QuČbec) Canada J2S 8E3 TČlČphone/Phone: 450-773-1105 TČlČcopieur/ Fax: 450-773-8461 montpetitd-at-agr.gc.ca
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Message d'origine----- DeÝ: acruz04-at-cibnor.mx [mailto:acruz04-at-cibnor.mx] EnvoyČÝ: jeudi 29 juin 2006 21:07 żÝ: Montpetit, Diane ObjetÝ: [Microscopy] viaWWW: Problem with EDX mail: acruz04-at-cibnor.mx Name: Ariel Cruz
Organization: CIBNOR SC
Title-Subject: [Filtered] Problem with EDX
Question: I really need your help it but quick possible, notice that working with the EDX Inca Oxford 2000 suddenly accent of receiving sign and it marked me overflow, to what tries to restart but it sends me the windows that I also send you (overflow, automation error that it lacks a file MCPDLL.DLL). Deadtime to 99%.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both dyel-at-mail.nih.gov as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dyel-at-mail.nih.gov Name: Chip Dye
Organization: NIH
Title-Subject: [Filtered] Flat embedding
Question: Dear ListServers:
Would anyone have any recommendations on the best way to flat embed mouse embryos (12 days old) which have been sectioned on a vibratome? I embedded the embryos in 10% gelatin prior to sectioning and cut 50 micron sections and embedded in Epon. It was during dehydration and infiltration, that my sections started to curl. After polymerizing in a flat mold, my sections looked like curled "potato chips". Would cutting the sections thicker help with this issue? Any assistance as to the best method to keep sections from doing this, would be greatly appreciated.
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Question: We are looking for manuals for an old controller and photomultiplier tube for a fluorescence spectroscopy system. The set up was used in the mid 90s and we are hoping to use it again but cannot find the manuals for these components.
The controller is an Action Research Corporation model number 275. The photomultiplier tube is from Research Inc. in Davers, MA model number R636/0115/0381.
Please let me know if anyone has copies of these manuals and we could pay for copying and shiping.
I used clean and subbed CR-39 plastic or glass 3x1" dlides and Spurr's resin embedding (or occasionally LR White I think). The Spurr's resin was removed by soaking the slides for around 20 minutes in sodium ethoxide (1.5 NaOH in 100cm3 ethanol and aged for 1 day prior to use). Always use freshly made sodium ethoxide (sodium chloride dissolved in ethanol). Never buy the ready made stuff and always make it up fresh after a few days use - although our less descerning histology lab kept there's a few weeks when batching slides through (and lost more sections). I very rarely, if ever lost the tissue section, but I always kept an eye on the slides as they were soaking (the resin starts to sag on the slide surface). Looking at them often helps agitate the solution (although with lung it's diffiicult to see if the section has fallen off until after washing - I can't remember what I washed the slides in but I assume it was neat ethanol, and possibly steps though serial rehydration (I didn't publish that bit - it must be in my lab notebooks in the attic). I used a standard (at the time) carousel plastic slide holder that could hold 50 slides in a large'ish beaker (Just Plastics plc). Our workshop made a PTFE ring to hold the slide carousel down as it had a tendency to float with a few glass or all CR-39 slides. CR-39 slides were used to create 235-UO2 fission frgament autoradiographs and glass slides (the serial section) were stained with H&E. This method doesn't 'etch' Spurr resin - I assume you want to completely remove from the section. We never had any residual Spurr's on the slide (it would have interfered with our 'soft alpha particle' shadowing technique used to visualise the tissue section into the CR-39 plastic and made us cross) .
Our (mainly lung) tissue was always fixed in freon FC-80 dissolved 1% Osmium tetroxide, followed by fixation in 3% gluteraldehyde storage in 0.1M sodium dicodylate buffer (4oC). Although fixing to EM standards for section quality, we viewed under the light microscope with a Magiscan Colour image analyser system.
Keith
------------------------------------------------ Dr Keith J Morris Cell Biology Division Institute of Ophthalmology University College London 11-43 bath Street London EC1V 9EL
Tel: 020 7608 4050
----- Original Message ----- X-from: {mayas003-at-yahoo.com} To: {keith.morris-at-ucl.ac.uk} Sent: Friday, June 30, 2006 5:31 PM
I need to replace my old critical point dryer and was wondering if anyone has any leads on the purchase of a previously owned one. Any assistance is much appreciated. Thanks.
Kelly
-- S. Kelly Sears, Ph.D., B.F.A. Facility for Electron Microscopy Research McGill University
==============================Original Headers============================== 6, 21 -- From sksears-at-eps.mcgill.ca Tue Jul 4 13:37:39 2006 6, 21 -- Received: from torrent.cc.mcgill.ca (torrent.CC.McGill.CA [132.206.27.49]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k64IbdVk015835 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Jul 2006 13:37:39 -0500 6, 21 -- Received: from mailscan5.CC.McGill.CA (mailscan5.CC.McGill.CA [132.216.77.252]) 6, 21 -- by torrent.cc.mcgill.ca (8.12.11/8.12.3) with ESMTP id k64Ibcch008649 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Jul 2006 14:37:38 -0400 6, 21 -- Received: from eps.mcgill.ca (dhcp21643226.Medicine.McGill.CA [132.216.43.226]) 6, 21 -- by mailscan5.CC.McGill.CA (8.12.11/8.12.11) with ESMTP id k64IbQdi031597 6, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 4 Jul 2006 14:37:27 -0400 6, 21 -- Message-ID: {44AAB56B.803-at-eps.mcgill.ca} 6, 21 -- Date: Tue, 04 Jul 2006 14:37:31 -0400 6, 21 -- From: "S. Kelly Sears" {sksears-at-eps.mcgill.ca} 6, 21 -- Organization: Facility for Electron Microscopy Research 6, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 6, 21 -- X-Accept-Language: en-us, en 6, 21 -- MIME-Version: 1.0 6, 21 -- To: Microscopy-at-microscopy.com 6, 21 -- Subject: Needed: Critical Point Dryer 6, 21 -- Content-Type: text/plain; charset=us-ascii; format=flowed 6, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am wondering if hydrofluoric acid technique to remove glass coverslips from cultured cell monolayers is compatible with LR White resin and subsequent immunolabeling. I mostly see in the Listserv archives references to Araldite or Embed 812. Any thoughts would be appreciated.
Best Regards, Kirk
Kirk J. Czymmek, Ph.D. Associate Professor Department of Biological Sciences University of Delaware Newark, DE 19716
==============================Original Headers============================== 5, 18 -- From kirk-at-UDel.Edu Wed Jul 5 08:00:54 2006 5, 18 -- Received: from copland.udel.edu (copland.udel.edu [128.175.13.92]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65D0snr009736 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 08:00:54 -0500 5, 18 -- Received: from [128.175.253.124] (host-253-124.nss.udel.edu [128.175.253.124]) 5, 18 -- by copland.udel.edu (8.13.6/8.13.6) with ESMTP id k65D0rrJ018283 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 09:00:53 -0400 (EDT) 5, 18 -- Message-ID: {44ABBAC1.7030305-at-udel.edu} 5, 18 -- Date: Wed, 05 Jul 2006 09:12:33 -0400 5, 18 -- From: kirk czymmek {kirk-at-UDel.Edu} 5, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.8) Gecko/20050511 5, 18 -- X-Accept-Language: en-us, en 5, 18 -- MIME-Version: 1.0 5, 18 -- To: Microscopy-at-microscopy.com 5, 18 -- Subject: Hydrofluoric acid and LR White 5, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 5, 18 -- Content-Transfer-Encoding: 7bit 5, 18 -- X-Scanned-By: MIMEDefang 2.52 on 128.175.13.92 ==============================End of - Headers==============================
} I am wondering if hydrofluoric acid technique to remove glass coverslips } from cultured cell monolayers is compatible with LR White resin and } subsequent immunolabeling. I mostly see in the Listserv archives references to Araldite or Embed 812. Any thoughts would be appreciated.
Dear Kirk, I just tried that very thing recently, with less-than-stellar results. The block faces were soft and difficult to cut. I got zero labelling, but I'm not sure what that was due to. I do know that my secondary antibodies were fine (I tested them on a known sample). Now, I must say that I do not know if the blocks were soft because of the HF treatment, or because they hadn't polymerized fully in the first place...there were some trapped air bubbles that may have interfered with it. In hindsight, my advise would be to make up a few blank, dummy blocks to test it out.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 4, 23 -- From lcgould-at-med.cornell.edu Wed Jul 5 08:41:16 2006 4, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65DfGEX020361 4, 23 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 08:41:16 -0500 4, 23 -- Received: from mpx2.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 4, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k65DfCRD026497 4, 23 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 09:41:13 -0400 (EDT) 4, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 4, 23 -- by mpx2.med.cornell.edu 4, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 4, 23 -- with ESMTPA id {0J1X00K5DNCN9X10-at-mpx2.med.cornell.edu} for 4, 23 -- microscopy-at-microscopy.com; Wed, 05 Jul 2006 09:41:12 -0400 (EDT) 4, 23 -- Date: Wed, 05 Jul 2006 09:33:40 -0400 4, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 4, 23 -- Subject: Re: [Microscopy] Hydrofluoric acid and LR White 4, 23 -- In-reply-to: {200607051302.k65D26In010993-at-ns.microscopy.com} 4, 23 -- Sender: lcgould-at-med.cornell.edu 4, 23 -- To: kirk-at-UDel.Edu, Microscopy Listserver {microscopy-at-microscopy.com} 4, 23 -- Message-id: {p06230904c0d16e45d76b-at-[140.251.48.23]} 4, 23 -- MIME-version: 1.0 4, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 4, 23 -- References: {200607051302.k65D26In010993-at-ns.microscopy.com} 4, 23 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.7.5.62432 ==============================End of - Headers==============================
I just share with you a trick I got from this list itself some time ago. I tried it with success and now I use it routinely. To obtain perfectly flat surface of resin-embedded cell monolayers, you just have to:
- Prepare open capsules by cutting off the dead end of beam capsules. - Grow your cells on coverslips - When it is time to embed, use just enough resin as to cover the coverslip with a few mm (not ml) resin (it is about 1-1.5 ml for a 3.5cm petri dish). Then return a beam capsule on your coverslip, press firmly so there is no space between the capsule and the coverslip. - Cure again. During the curing, the capsule will be glued in the resin. Next day fill the capsule with fresh resin and cure again (from my own experience doubling the time of curing help detaching the blocks from the coverslips). - When the capsules are well cured, use a pair of forceps to take the capsule off the coverslip. Sometimes some of the capsule surface sticks to the coverslips but you usually always keep enough surface to be ultracut. "overcuring" really helps here. You can also use 4 capsules per coverslip, which gives you complety security (I personally use 2 capsules per coverslip and usually don't need the second). - The surface of the block is perfectly flat, the first section will be good!! (which is interesting if you have very flat cells like me)
Good luck
Stéphane
--- kirk-at-UDel.Edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Microscopy-at-microscopy.com } } Dear Microscopists, } } I am wondering if hydrofluoric acid technique to } remove glass coverslips } from cultured cell monolayers is compatible with LR } White resin and } subsequent immunolabeling. I mostly see in the } Listserv archives } references to Araldite or Embed 812. Any thoughts } would be appreciated. } } Best Regards, Kirk } } Kirk J. Czymmek, Ph.D. } Associate Professor } Department of Biological Sciences } University of Delaware } Newark, DE 19716 } } ==============================Original } Headers============================== } 5, 18 -- From kirk-at-UDel.Edu Wed Jul 5 08:00:54 2006 } 5, 18 -- Received: from copland.udel.edu } (copland.udel.edu [128.175.13.92]) } 5, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k65D0snr009736 } 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 } Jul 2006 08:00:54 -0500 } 5, 18 -- Received: from [128.175.253.124] } (host-253-124.nss.udel.edu [128.175.253.124]) } 5, 18 -- by copland.udel.edu (8.13.6/8.13.6) with } ESMTP id k65D0rrJ018283 } 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 5 } Jul 2006 09:00:53 -0400 (EDT) } 5, 18 -- Message-ID: {44ABBAC1.7030305-at-udel.edu} } 5, 18 -- Date: Wed, 05 Jul 2006 09:12:33 -0400 } 5, 18 -- From: kirk czymmek {kirk-at-UDel.Edu} } 5, 18 -- User-Agent: Mozilla/5.0 (Windows; U; } Windows NT 5.0; en-US; rv:1.7.8) Gecko/20050511 } 5, 18 -- X-Accept-Language: en-us, en } 5, 18 -- MIME-Version: 1.0 } 5, 18 -- To: Microscopy-at-microscopy.com } 5, 18 -- Subject: Hydrofluoric acid and LR White } 5, 18 -- Content-Type: text/plain; charset=us-ascii; } format=flowed } 5, 18 -- Content-Transfer-Encoding: 7bit } 5, 18 -- X-Scanned-By: MIMEDefang 2.52 on } 128.175.13.92 } ==============================End of - } Headers============================== }
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I would assume that overcuring or increased embedding could well interfere with the immunolabelling.
So I can only suggest trying liquid nitrogen to free the coverslips or else using an alternative such as melinex instead of glass coverslips because that can be sectioned if necessary. I cannot speak from experience about their use in immunolabelling but have used both techniques as an alternative to HF.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: nizets2-at-yahoo.com
Hi all
I need a software to build a panorama image, or geant image from a set of overlapping images taken by SEM.
On the web one can find dozen of freewares, sharewares or commercialy one, and I used one which works great, but they are all under windows. The only one I found running under linux is "PanTools" which should work on all OS, but I don't catch how to install and use it. It should work as a plugin for gimp, but... I tried an ImageJ plugin too, but the most part was to be done manually.
So any suggestions are welcome.
-- J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322
-----Original Message----- X-from: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Friday, June 16, 2006 2:07 PM To: lesley.bechtold-at-jax.org
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I know Autostitch doesn't fall under the 'linux' OS, but its been *THE* best far and beyond for putting together images (in my experience).
Well worth the hassle to move to a windows platform in most cases I would guess.
*I do not have any financial/developmental interest in the software, no connection to Matthew Brown. I am just a very satisfied user of the software. Web page is www.cs.ubc.ca/~mbrown/autostitch/autostitch.html
Regards,
Geoff Williams Leduc Bioimaging Facility Manager Brown University
http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/ -----Original Message----- X-from: jacques.faerber-at-ipcms.u-strasbg.fr [mailto:jacques.faerber-at-ipcms.u-strasbg.fr] Sent: Wednesday, July 05, 2006 12:00 PM To: Williams, Geoffrey
Hi all
I need a software to build a panorama image, or geant image from a set of overlapping images taken by SEM.
On the web one can find dozen of freewares, sharewares or commercialy one, and I used one which works great, but they are all under windows. The only one I found running under linux is "PanTools" which should work on all OS, but I don't catch how to install and use it. It should work as a plugin for gimp, but... I tried an ImageJ plugin too, but the most part was to be done manually.
So any suggestions are welcome.
-- J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
On Jul 5, 2006, at 8:59 AM, jacques.faerber-at-ipcms.u-strasbg.fr wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Hi all } } I need a software to build a panorama image, or geant image from a set } of overlapping images taken by SEM. } } On the web one can find dozen of freewares, sharewares or commercialy } one, and I used one which works great, but they are all under } windows. } The only one I found running under linux is "PanTools" which should } work } on all OS, but I don't catch how to install and use it. It should work } as a plugin for gimp, but... } I tried an ImageJ plugin too, but the most part was to be done } manually. } } So any suggestions are welcome. } } } -- } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } } ==============================Original } Headers============================== } 8, 28 -- From jacques.faerber-at-ipcms.u-strasbg.fr Wed Jul 5 } 10:55:31 2006 } 8, 28 -- Received: from mailhost.u-strasbg.fr (mailhost.u- } strasbg.fr [130.79.200.157]) } 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k65FtU0M021246 } 8, 28 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 10:55:31 } -0500 } 8, 28 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr } [130.79.210.2]) } 8, 28 -- by mailhost.u-strasbg.fr (8.13.6/jtpda-5.5pre1) } with ESMTP id k65FtUTD029465 } 8, 28 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 } 17:55:30 +0200 (CEST) } 8, 28 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr } [130.79.54.3]) } 8, 28 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id B76F710000E4 } 8, 28 -- for {Microscopy-at-Microscopy.Com} ; Wed, 5 Jul 2006 } 17:55:22 +0200 (CEST) } 8, 28 -- Message-ID: {44ABE0D2.9070800-at-ipcms.u-strasbg.fr} } 8, 28 -- Date: Wed, 05 Jul 2006 17:54:58 +0200 } 8, 28 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} } 8, 28 -- User-Agent: Mozilla Thunderbird 1.0.8 (X11/20060503) } 8, 28 -- X-Accept-Language: fr, en } 8, 28 -- MIME-Version: 1.0 } 8, 28 -- To: Microscopy-at-microscopy.com } 8, 28 -- Subject: panorama software under linux } 8, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 8, 28 -- Content-Transfer-Encoding: 8bit } 8, 28 -- X-IPCMS-MailScanner: Found to be clean } 8, 28 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr } 8, 28 -- X-Greylist: Sender IP whitelisted, not delayed by milter- } greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.157]); Wed, 05 } Jul 2006 17:55:30 +0200 (CEST) } 8, 28 -- X-Virus-Scanned: ClamAV 0.88.2/1585/Tue Jul 4 22:39:34 } 2006 on mr7.u-strasbg.fr } 8, 28 -- X-Virus-Status: Clean } 8, 28 -- X-Spam-Status: No, score=-1.4 required=5.0 } tests=ALL_TRUSTED,AWL } 8, 28 -- autolearn=disabled version=3.1.1 } 8, 28 -- X-Spam-Checker-Version: SpamAssassin 3.1.1 (2006-03-10) on } mr7.u-strasbg.fr } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 23 -- From Elliott-at-arizona.edu Wed Jul 5 13:39:15 2006 9, 23 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65IdEnG031716 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 13:39:14 -0500 9, 23 -- Received: from localhost (faramir.email.arizona.edu [10.0.0.218]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id E2BF5E95932 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 11:39:02 -0700 (MST) 9, 23 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 268BBE9726A 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 11:38:51 -0700 (MST) 9, 23 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 23 -- In-Reply-To: {200607051559.k65FxFOj026814-at-ns.microscopy.com} 9, 23 -- References: {200607051559.k65FxFOj026814-at-ns.microscopy.com} 9, 23 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 23 -- Message-Id: {A38537E5-E446-4194-8DA4-37EC4DA2FBF1-at-arizona.edu} 9, 23 -- From: David Elliott {Elliott-at-arizona.edu} 9, 23 -- Subject: Re: [Microscopy] panorama software under linux 9, 23 -- Date: Wed, 5 Jul 2006 11:38:50 -0700 9, 23 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 9, 23 -- X-Mailer: Apple Mail (2.752.2) 9, 23 -- X-Virus-Scanned: amavisd-new at email.arizona.edu 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k65IdEnG031716 ==============================End of - Headers==============================
We routinely embed "Vibratome" sections between two pieces of ACLAR cut to fit on a standard 1" X 3" microscope slide (For convenience in handling--smaller or larger pieces can be used). This does not always result in a perfectly flat sample. If that is a problem then a small weight can be placed on top of the sample during polymerization. This may make a very fragile sample so just carefully remove one of the ACLAR pieces (I use a razor blade to separate the two pieces of ACLAR and then gently pull them apart) and add a few drops of epoxy to the sample and polymerize. This will give a sturdy sample that is very flat on one side. If you wish to section a small part of your sample cut out the area with a razor blade and glue (with cyanoacrylate "SuperGlue") or use epoxy resin to attach to a blank specimen block. Leave a little margin around the area of interest when cutting a sample with a razor blade because the edges may curl a bit during the cutting/gluing process.
Another quick solution if you do not have ACLAR on hand: Make some very thin epoxy sheets by spreading epoxy resin very thinly on your embedding molds. You can even use the backside of the molds to get larger pieces. Polymerization need not be complete but, for example, 24 hours of a 48 hour process. Cut out two pieces and embedd your tissue section with a couple drops of resin between them. Polymerize.
Cutting thicker sections will reduce the curling during processing but not eliminate it util you get to one millimeter or so which makes light microscopy and trimming specific areas of tissue a big problem. I have embedded many tissues, from drosophila embryos to one centimeter diameter sections of mamalian brain, with this basic technique. Best wishes Larry
dyel-at-mail.nih.gov wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both dyel-at-mail.nih.gov as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: dyel-at-mail.nih.gov } Name: Chip Dye } } Organization: NIH } } Title-Subject: [Filtered] Flat embedding } } Question: Dear ListServers: } } Would anyone have any recommendations on the best way to flat embed } mouse embryos (12 days old) which have been sectioned on a vibratome? } I embedded the embryos in 10% gelatin prior to sectioning and cut 50 } micron sections and embedded in Epon. It was during dehydration and } infiltration, that my sections started to curl. After polymerizing } in a flat mold, my sections looked like curled "potato chips". Would } cutting the sections thicker help with this issue? Any assistance as } to the best method to keep sections from doing this, would be greatly } appreciated. } } Regards, } } Chip Dye } } Microscopist } Microscopy & Imaging Core, NICHD, NIH } Building 49, Room 5W-14 } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } Phone: 301-496-3627 } E-mail: dyel-at-mail.nih.gov } } } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Sat Jul 1 05:34:49 2006 } 12, 12 -- Received: from [192.169.1.141] (msdvpn24.msd.anl.gov [130.202.238.88]) } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k61AYk3B004150 } 12, 12 -- for {microscopy-at-microscopy.com} ; Sat, 1 Jul 2006 05:34:48 -0500 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p06020401c0cc003743f2-at-[192.169.1.141]} } 12, 12 -- Date: Sat, 1 Jul 2006 05:34:46 -0500 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: dyel-at-mail.nih.gov (by way of MicroscopyListserver) } 12, 12 -- Subject: viaWWW: Flat embedding } 12, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
All: the URL I previous sent has no spaces in it. Must have been a finger mis-poke. http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTable/
r-holdford-at-ti.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Debby and anyone else who wants it: here's a link to download the files: } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTable/ } } Debby Sherman wrote: } } } Gee..would you mind sharing that program with others? I would love to have } } a copy. } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } http://www.agriculture.purdue.edu/microscopy } } } } } } On 6/28/06 6:37 PM, "r-holdford-at-ti.com" {r-holdford-at-ti.com} wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } } } Owen: one of my favorites is EM Periodic Table, a program authored by } } } Scott Walck many years ago. It has a periodic chart layout and a table } } } layout that can be translated to Excel. I got it at a Lehigh short } } } course and I find it very handy. I'd be happy to share it with you. } } } Scott calls this "beerware" meaning when we meet him at conferences, we } } } owe him a beer if we use it. } } } } } } Scott: you can claim your pitcher at M&M in Chicago. This program } } } would be cheap at twice the price. } } } } } } opmills-at-mtu.edu wrote: } } } } } } } } } } ---------------------------------------------------------------------------- } } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } ---------------------------------------------------------------------------- } } } } } } } } Does anyone know where I can find a spreadsheet or text version of x- } } } } ray emission lines? } } } } } } } } Thanks, } } } } } } } } OWen } } } } } } } } } } } } } } } } } } } } Owen P. Mills } } } } Director, Materials Characterization & Fabrication Facilities } } } } Electron Optics Engineer, Applied Chemical & Morphological Analysis } } } } Laboratory } } } } } } } } Materials Science & Engineering } } } } Michigan Technological University } } } } Rm 512 M&M Bldg. } } } } Houghton, MI 49931 } } } } PH 906-369-1875 } } } } FAX 906-487-2934 } } } } mailto:opmills-at-mtu.edu } } } } http://www.mm.mtu.edu/~opmills } } } } } } } } } } } } } } } } ==============================Original Headers============================== } } } } 10, 31 -- From opmills-at-mtu.edu Wed Jun 28 14:59:39 2006 } } } } 10, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) } } } } 10, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } } } k5SJxdNZ021212 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 14:59:39 -0500 } } } } 10, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu } } } } [141.219.69.6]) } } } } 10, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id k5SJxc9N010211 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } } } } 10, 31 -- Received: from node2.edge.dcsint.mtu.edu (node2.mtu.edu } } } } [141.219.69.2]) } } } } 10, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP } } } } id k5SJxcCb016435 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } } } } 10, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) } } } } 10, 31 -- by node2.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id } } } } k5SJxbZH003257 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:38 -0400 } } } } 10, 31 -- (envelope-from opmills-at-mtu.edu) } } } } 10, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu } } } } [141.219.69.6]) } } } } 10, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k5SJxbQP012706 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:37 -0400 } } } } (EDT) } } } } 10, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu } } } } [141.219.192.45]) } } } } 10, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc } } } } v1.0) with ESMTP id k5SJxbbF016425 } } } } 10, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 28 Jun 2006 15:59:37 -0400 } } } } 10, 31 -- Mime-Version: 1.0 (Apple Message framework v750) } } } } 10, 31 -- Content-Transfer-Encoding: 7bit } } } } 10, 31 -- Message-Id: {82FC92F2-DB6B-4300-BE2B-59C95CA02CBB-at-mtu.edu} } } } } 10, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } } } } format=flowed } } } } 10, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} } } } } 10, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} } } } } 10, 31 -- Subject: need x-ray emission table } } } } 10, 31 -- Date: Wed, 28 Jun 2006 15:59:46 -0400 } } } } 10, 31 -- X-Mailer: Apple Mail (2.750) } } } } 10, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.4.0.264935, } } } } Antispam-Data: 2006.6.28.123433 } } } } 10, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 } } } } 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, } } } } __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' } } } } ==============================End of - Headers============================== } } } } } } } } } } } } } } } } } } } } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 22 -- From r-holdford-at-ti.com Wed Jul 5 14:50:15 2006 4, 22 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65JoDD5020806 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 14:50:15 -0500 4, 22 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 22 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k65Jo8CS019260 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 14:50:13 -0500 (CDT) 4, 22 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 22 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k65Jo7GA024590 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 14:50:07 -0500 (CDT) 4, 22 -- Message-ID: {44AC17EF.4050803-at-ti.com} 4, 22 -- Date: Wed, 05 Jul 2006 14:50:07 -0500 4, 22 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 22 -- Organization: SC Packaging Development -- FA Development 4, 22 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 4, 22 -- MIME-Version: 1.0 4, 22 -- To: MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Subject: Re: [Microscopy] need x-ray emission table 4, 22 -- References: {200607051814.k65IEexl011744-at-ns.microscopy.com} 4, 22 -- In-Reply-To: {200607051814.k65IEexl011744-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
The issue of flat-embedding has been addressed by several authors, including Schwartz (1982), Schafer (1989), Nguyen and Pender (1995), Larue and Winer (1996). In my processing I largely follow the approach established by Larue and Winer (1996) with some modifications (Larue DT, Winer JA. Postembedding immunocytochemistry of large sections of brain tissue: an improved flat embedding technique. J Neurosci Methods, 1996 Sep; 68(1):125-132). I work with the 50 micron thick vibratome sections of mouse brain embedded in agar which for me works better then gelatin. Normally, for flat embedding I osmicate and/or dehydrate the vibratome sections placed between two membrane filters disks with diameter at least two-three times exceeding the tissue size. The sections remained flat between two membrane disks due to the surface tension. The severe section distortion during dehydration is avoided or at least minimized.
The main disadvantage is the increased number of fluid changes and extended dehydration times compared to routine dehydration protocols, which makes sample preparation relatively time consuming. Briefly: place disk of filter paper into wetted compartment of Polystyrene multidish plate, transfer fixed vibratome sections onto filter disk and cover with the other one. Carefully add the solution not allowing the top filter to float. The challenge is to keep a certain minimal solution volume to maintain a surface tension between filters and yet sufficient to avoid sample drying. I not always use propylene oxide when working with TAAB embedding and the polystyrene plate works well with ethanol. For the last 100% change I add alcohol in excess to remove top filter disc without breaking the section. Aclar or Kapton films sandwich works very well for the polymerization Hope this helps, Albina
On Sat Jul 01 06:45:28 EDT 2006, dyel-at-mail.nih.gov wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so } when replying } please copy both dyel-at-mail.nih.gov as well as the MIcroscopy } Listserver } --------------------------------------------------------------------------- } } Email: dyel-at-mail.nih.gov } Name: Chip Dye } } Organization: NIH } } Title-Subject: [Filtered] Flat embedding } } Question: Dear ListServers: } } Would anyone have any recommendations on the best way to flat } embed mouse embryos (12 days old) which have been sectioned on a } vibratome? I embedded the embryos in 10% gelatin prior to } sectioning and cut 50 micron sections and embedded in Epon. It } was during dehydration and infiltration, that my sections started } to curl. After polymerizing in a flat mold, my sections looked } like curled "potato chips". Would cutting the sections thicker } help with this issue? Any assistance as to the best method to } keep sections from doing this, would be greatly appreciated. } } Regards, } } Chip Dye } } Microscopist } Microscopy & Imaging Core, NICHD, NIH } Building 49, Room 5W-14 } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } Phone: 301-496-3627 } E-mail: dyel-at-mail.nih.gov } } } } --------------------------------------------------------------------------- } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Sat Jul 1 05:34:49 2006 } 12, 12 -- Received: from [192.169.1.141] (msdvpn24.msd.anl.gov } [130.202.238.88]) } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k61AYk3B004150 } 12, 12 -- for {microscopy-at-microscopy.com} ; Sat, 1 Jul 2006 } 05:34:48 -0500 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p06020401c0cc003743f2-at-[192.169.1.141]} } 12, 12 -- Date: Sat, 1 Jul 2006 05:34:46 -0500 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: dyel-at-mail.nih.gov (by way of } MicroscopyListserver) } 12, 12 -- Subject: viaWWW: Flat embedding } 12, 12 -- Content-Type: text/plain; charset="us-ascii" ; } format="flowed" } ==============================End of - } Headers============================== } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 8, 22 -- From amich-at-ufl.edu Wed Jul 5 15:49:07 2006 8, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65Kn6EB031660 8, 22 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 15:49:07 -0500 8, 22 -- Received: from osgjas03.cns.ufl.edu (osgjas03.cns.ufl.edu [128.227.74.133]) 8, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k65Kn0NW4886700; 8, 22 -- Wed, 5 Jul 2006 16:49:01 -0400 8, 22 -- Message-ID: {2028944878.112041152132540772.JavaMail.osg-at-osgjas03.cns.ufl.edu} 8, 22 -- Date: Wed, 5 Jul 2006 16:49:00 -0400 (EDT) 8, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 8, 22 -- To: dyel-at-mail.nih.gov 8, 22 -- Subject: Re: [Microscopy] viaWWW: Flat embedding 8, 22 -- Cc: microscopy-at-microscopy.com 8, 22 -- MIME-Version: 1.0 8, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 8, 22 -- Content-Transfer-Encoding: 7bit 8, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 8, 22 -- X-Originating-IP: 70.152.43.35 [70.152.43.35] 8, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 8, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 8, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Ritchie, Geoff and others: for some odd reason my email program is thinking there is a space when there isn't one, so the link is not finished. If you copy and paste the URL into your browser it seems to work. Check to see if there is a space in the copy before you press 'enter'. There should be NO spaces in the URL.
Ritchie Sims wrote: } Hi } } Still doesn't work, and looks the same, did you do the same mis-poke? } } please try again } } cheers } } rtch } } } } On 5 Jul 2006 at 14:51, r-holdford-at-ti.com wrote: } } } } } } ---------------------------------------------------------------------- } } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver On-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } ------ } } } } All: the URL I previous sent has no spaces in it. Must have been a } } finger mis-poke. } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } able/ } } } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 23 -- From r-holdford-at-ti.com Wed Jul 5 15:52:30 2006 4, 23 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65KqUUj004901 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 15:52:30 -0500 4, 23 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) 4, 23 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k65Kq75D016380; 4, 23 -- Wed, 5 Jul 2006 15:52:12 -0500 (CDT) 4, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 23 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k65Kq6Pu021536; 4, 23 -- Wed, 5 Jul 2006 15:52:06 -0500 (CDT) 4, 23 -- Message-ID: {44AC2676.1000800-at-ti.com} 4, 23 -- Date: Wed, 05 Jul 2006 15:52:06 -0500 4, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 23 -- Organization: SC Packaging Development -- FA Development 4, 23 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 4, 23 -- MIME-Version: 1.0 4, 23 -- To: Ritchie Sims {r.sims-at-auckland.ac.nz} , mcauliff-at-umdnj.edu, 4, 23 -- MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 23 -- Subject: Re: [Microscopy] Re: need x-ray emission table 4, 23 -- References: {44ACC831.12352.4E962-at-localhost} 4, 23 -- In-Reply-To: {44ACC831.12352.4E962-at-localhost} 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Just as an FYI, the first and second tries worked fine for me... I didn't have any problem with the url sent.
dj
On Wed, 5 Jul 2006, r-holdford-at-ti.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Ritchie, Geoff and others: for some odd reason my email program is } thinking there is a space when there isn't one, so the link is not } finished. If you copy and paste the URL into your browser it seems to } work. Check to see if there is a space in the copy before you press } 'enter'. There should be NO spaces in the URL. } } Ritchie Sims wrote: } } Hi } } } } Still doesn't work, and looks the same, did you do the same mis-poke? } } } } please try again } } } } cheers } } } } rtch } } } } } } } } On 5 Jul 2006 at 14:51, r-holdford-at-ti.com wrote: } } } } } } } } } } ---------------------------------------------------------------------- } } } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } } } of America To Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserver On-Line Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } } } ------ } } } } } } All: the URL I previous sent has no spaces in it. Must have been a } } } finger mis-poke. } } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } } able/ } } } } } } } } } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-995-2360 } 972-648-8743 (pager) } SC Packaging FA Development } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } ==============================Original Headers============================== } 4, 23 -- From r-holdford-at-ti.com Wed Jul 5 15:52:30 2006 } 4, 23 -- Received: from reloaded.ext.ti.com (reloaded.ext.ti.com [192.94.94.6]) } 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65KqUUj004901 } 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 5 Jul 2006 15:52:30 -0500 } 4, 23 -- Received: from dlep31.itg.ti.com ([157.170.170.35]) } 4, 23 -- by reloaded.ext.ti.com (8.13.4/8.13.4) with ESMTP id k65Kq75D016380; } 4, 23 -- Wed, 5 Jul 2006 15:52:12 -0500 (CDT) } 4, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) } 4, 23 -- by dlep31.itg.ti.com (8.12.11/8.12.11) with ESMTP id k65Kq6Pu021536; } 4, 23 -- Wed, 5 Jul 2006 15:52:06 -0500 (CDT) } 4, 23 -- Message-ID: {44AC2676.1000800-at-ti.com} } 4, 23 -- Date: Wed, 05 Jul 2006 15:52:06 -0500 } 4, 23 -- From: Becky Holdford {r-holdford-at-ti.com} } 4, 23 -- Organization: SC Packaging Development -- FA Development } 4, 23 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) } 4, 23 -- MIME-Version: 1.0 } 4, 23 -- To: Ritchie Sims {r.sims-at-auckland.ac.nz} , mcauliff-at-umdnj.edu, } 4, 23 -- MSA ListServer {Microscopy-at-MSA.Microscopy.Com} } 4, 23 -- Subject: Re: [Microscopy] Re: need x-ray emission table } 4, 23 -- References: {44ACC831.12352.4E962-at-localhost} } 4, 23 -- In-Reply-To: {44ACC831.12352.4E962-at-localhost} } 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 23 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 5, 19 -- From dljones-at-bestweb.net Wed Jul 5 16:01:40 2006 5, 19 -- Received: from vms048pub.verizon.net (vms048pub.verizon.net [206.46.252.48]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k65L1erQ019493 5, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 16:01:40 -0500 5, 19 -- Received: from localhost ([70.107.112.47]) 5, 19 -- by vms048.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 5, 19 -- 9 2005)) with ESMTPA id {0J1Y00CB77Q24RD9-at-vms048.mailsrvcs.net} for 5, 19 -- Microscopy-at-microscopy.com; Wed, 05 Jul 2006 16:01:19 -0500 (CDT) 5, 19 -- Date: Wed, 05 Jul 2006 17:12:32 -0400 (Eastern Standard Time) 5, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 5, 19 -- Subject: Re: [Microscopy] need x-ray emission table 5, 19 -- In-reply-to: {200607052056.k65KuXxo016061-at-ns.microscopy.com} 5, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 5, 19 -- To: r-holdford-at-ti.com 5, 19 -- Cc: Microscopy-at-microscopy.com 5, 19 -- Message-id: {Pine.WNT.4.64.0607051711330.4076-at-H-F1} 5, 19 -- MIME-version: 1.0 5, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 5, 19 -- References: {200607052056.k65KuXxo016061-at-ns.microscopy.com} ==============================End of - Headers==============================
The fatty and relatively soft brain tissue I embed in 3- 5% low melting point agarose (Sigma Chemical Co., cat# A-9414). I prefer not to trim the block too close to tissue: it seems that excess agarose helping prevent sections from curling.
} What percentage agar do you use when making the brain sections? } Is this regular bacteriological agar or low melting agarose? I } am about to embark on a similar project. I was thinking of } putting the sections into mesh biopsy bags to keep them flat. } } amich-at-ufl.edu wrote: } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } The issue of flat-embedding has been addressed by several } } authors, including Schwartz (1982), Schafer (1989), Nguyen and } } Pender (1995), Larue and Winer (1996). In my processing I } } largely follow the approach established by Larue and Winer } } (1996) with some modifications (Larue DT, Winer JA. } } Postembedding immunocytochemistry of large sections of brain } } tissue: an improved flat embedding technique. J Neurosci } } Methods, 1996 Sep; 68(1):125-132). I work with the 50 micron } } thick vibratome sections of mouse brain embedded in agar which } } for me works better then gelatin. Normally, for flat embedding I } } osmicate and/or dehydrate the vibratome sections placed between } } two membrane filters disks with diameter at least two-three } } times exceeding the tissue size. The sections remained flat } } between two membrane disks due to the surface tension. The } } severe section distortion during dehydration is avoided or at } } least minimized. } } } } The main disadvantage is the increased number of fluid changes } } and extended dehydration times compared to routine dehydration } } protocols, which makes sample preparation relatively time } } consuming. Briefly: place disk of filter paper into wetted } } compartment of Polystyrene multidish plate, transfer fixed } } vibratome sections onto filter disk and cover with the other } } one. Carefully add the solution not allowing the top filter to } } float. The challenge is to keep a certain minimal solution } } volume to maintain a surface tension between filters and yet } } sufficient to avoid sample drying. I not always use propylene } } oxide when working with TAAB embedding and the polystyrene plate } } works well with ethanol. For the last 100% change I add alcohol } } in excess to remove top filter disc without breaking the section. } } Aclar or Kapton films sandwich works very well for the } } polymerization } } Hope this helps, } } Albina } } } } On Sat Jul 01 06:45:28 EDT 2006, dyel-at-mail.nih.gov wrote: } } } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society } } } of America } } } To Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } } } This Question/Comment was submitted to the Microscopy Listserver } } } using the WWW based Form at } } } http://microscopy.com/MicroscopyListserver/MLFormMail.html } } } --------------------------------------------------------------------------- } } } Remember this posting is most likely not from a Subscriber, so } } } when replying } } } please copy both dyel-at-mail.nih.gov as well as the } } } MIcroscopy Listserver } } } --------------------------------------------------------------------------- } } } } } } Email: dyel-at-mail.nih.gov } } } Name: Chip Dye } } } } } } Organization: NIH } } } } } } Title-Subject: [Filtered] Flat embedding } } } } } } Question: Dear ListServers: } } } } } } Would anyone have any recommendations on the best way to flat } } } embed mouse embryos (12 days old) which have been sectioned on } } } a vibratome? I embedded the embryos in 10% gelatin prior to } } } sectioning and cut 50 micron sections and embedded in Epon. It } } } was during dehydration and infiltration, that my sections } } } started to curl. After polymerizing in a flat mold, my } } } sections looked like curled "potato chips". Would cutting the } } } sections thicker help with this issue? Any assistance as to } } } the best method to keep sections from doing this, would be } } } greatly appreciated. } } } } } } Regards, } } } } } } Chip Dye } } } } } } Microscopist } } } Microscopy & Imaging Core, NICHD, NIH } } } Building 49, Room 5W-14 } } } 49 Convent Drive, MSC 4495, Bethesda, MD, 20892-4495 } } } Phone: 301-496-3627 } } } E-mail: dyel-at-mail.nih.gov } } } } } } } } } } } } --------------------------------------------------------------------------- } } } ==============================Original } } } Headers============================== } } } 12, 12 -- From zaluzec-at-microscopy.com Sat Jul 1 05:34:49 2006 } } } 12, 12 -- Received: from [192.169.1.141] (msdvpn24.msd.anl.gov } } } [130.202.238.88]) } } } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } } ESMTP id k61AYk3B004150 } } } 12, 12 -- for {microscopy-at-microscopy.com} ; Sat, 1 Jul 2006 } } } 05:34:48 -0500 } } } 12, 12 -- Mime-Version: 1.0 } } } 12, 12 -- X-Sender: (Unverified) } } } 12, 12 -- Message-Id: {p06020401c0cc003743f2-at-[192.169.1.141]} } } } 12, 12 -- Date: Sat, 1 Jul 2006 05:34:46 -0500 } } } 12, 12 -- To: microscopy-at-microscopy.com } } } 12, 12 -- From: dyel-at-mail.nih.gov (by way of } } } MicroscopyListserver) } } } 12, 12 -- Subject: viaWWW: Flat embedding } } } 12, 12 -- Content-Type: text/plain; charset="us-ascii" ; } } } format="flowed" } } } ==============================End of - } } } Headers============================== } } } } } } } } } } } } } } -- } } MIKHAYLOVA,ALBINA, PhD } } Post Doctoral Research Associate } } Materials Science and Engineering } } University of Florida } } } } ==============================Original } } Headers============================== } } 8, 22 -- From amich-at-ufl.edu Wed Jul 5 15:49:07 2006 } } 8, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu } } [128.227.74.149]) } } 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k65Kn6EB031660 } } 8, 22 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 } } 15:49:07 -0500 } } 8, 22 -- Received: from osgjas03.cns.ufl.edu } } (osgjas03.cns.ufl.edu [128.227.74.133]) } } 8, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id } } k65Kn0NW4886700; } } 8, 22 -- Wed, 5 Jul 2006 16:49:01 -0400 } } 8, 22 -- Message-ID: } } {2028944878.112041152132540772.JavaMail.osg-at-osgjas03.cns.ufl.edu} } } 8, 22 -- Date: Wed, 5 Jul 2006 16:49:00 -0400 (EDT) } } 8, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} } } 8, 22 -- To: dyel-at-mail.nih.gov } } 8, 22 -- Subject: Re: [Microscopy] viaWWW: Flat embedding } } 8, 22 -- Cc: microscopy-at-microscopy.com } } 8, 22 -- MIME-Version: 1.0 } } 8, 22 -- Content-Type: text/plain; format=flowed; } } charset=us-ascii } } 8, 22 -- Content-Transfer-Encoding: 7bit } } 8, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) } } 8, 22 -- X-Originating-IP: 70.152.43.35 [70.152.43.35] } } 8, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED } } 8, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, } } tests=ALL_TRUSTED } } 8, 22 -- X-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } 8, 22 -- X-UFL-Scanned-By: CNS Open Systems Group } } (http://open-systems.ufl.edu/services/smtp-relay/) } } ==============================End of - } } Headers============================== } } } } -- Greg Erdos } 5410 SE 185th Ave. } Micanopy, FL 32667 } }
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 7, 21 -- From amich-at-ufl.edu Wed Jul 5 21:10:02 2006 7, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k662A1pn001133 7, 21 -- for {microscopy-at-microscopy.com} ; Wed, 5 Jul 2006 21:10:02 -0500 7, 21 -- Received: from osgjas03.cns.ufl.edu (osgjas03.cns.ufl.edu [128.227.74.133]) 7, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k6629vkq1925370; 7, 21 -- Wed, 5 Jul 2006 22:09:58 -0400 7, 21 -- Message-ID: {4879372.115621152151797443.JavaMail.osg-at-osgjas03.cns.ufl.edu} 7, 21 -- Date: Wed, 5 Jul 2006 22:09:57 -0400 (EDT) 7, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 7, 21 -- To: gwe-at-ufl.edu, microscopy-at-microscopy.com 7, 21 -- Subject: Re: [Microscopy] Re: viaWWW: Flat embedding 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 7, 21 -- X-Originating-IP: 68.214.118.120 [68.214.118.120] 7, 21 -- X-Spam-Status: hits=0, required=5, tests= 7, 21 -- X-UFL-Spam-Status: hits=0, required=5, tests= 7, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 7, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Has anybody ever Experimentally tried this inside Wehnelt gold coating and measured or noticed a difference in the false peak response as an increase or decrease in the peak-to-peak amplitude of the false peak as seen on an older SEM's CRT waveform? (All other factors being equal.)
Has anyone ever Experimentally tried it with a thick carbon coating and noticed the peak-peak amplitude response of the false peak?
Paul
At 09:07 AM 6/24/06 -0500, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 25 -- From beaurega-at-westol.com Thu Jul 6 01:58:48 2006 7, 25 -- Received: from smtp-gateway-1.winbeam.com (smtp-gateway-1.winbeam.com [64.84.97.66]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k666wmmI020291 7, 25 -- for {microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 01:58:48 -0500 7, 25 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 7, 25 -- by smtp-gateway-1.winbeam.com (8.13.1/8.12.8) with SMTP id k666whiT003030 7, 25 -- for {microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 02:58:45 -0400 7, 25 -- Received: (qmail 31079 invoked by uid 89); 6 Jul 2006 06:58:41 -0000 7, 25 -- Received: from pitts-69-72-13-120.dynamic-dialup.coretel.net (HELO millenium) (69.72.13.120) 7, 25 -- by mail.winbeam.com with SMTP; 6 Jul 2006 06:58:41 -0000 7, 25 -- Message-Id: {3.0.6.32.20060706030001.007d46b0-at-pop3.norton.antivirus} 7, 25 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 7, 25 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 7, 25 -- Date: Thu, 06 Jul 2006 03:00:01 -0400 7, 25 -- To: microscopy-at-microscopy.com 7, 25 -- From: Beaurega {beaurega-at-westol.com} 7, 25 -- Subject: Re: [Microscopy] What would happen if ...... sputter coating 7, 25 -- Wehnelt assembly 7, 25 -- Mime-Version: 1.0 7, 25 -- Content-Type: text/plain; charset="us-ascii" 7, 25 -- X-Winbeam-MailScanner-Information: smtp-gateway-1.winbeam.com - Please contact Technical Support for more information 7, 25 -- X-Winbeam-MailScanner: Found to be clean [smtp-gateway-1.winbeam.com] (courtesy of Winbeam) 7, 25 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, score=0, 7, 25 -- required 6, autolearn=disabled) 7, 25 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
I am seriously considering investing in a microwave for our cell monolayers processing for TEM. I would like to have some comments from people already using this technique (advantages/disadvantages, worth the investment...) and also an idea of the money I would have to spend for it.
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
We use microwaves as our standard processing method now, both for decreasing our processing times and for improved quality of ultrastructure. We find that microwave processing requires much less time than conventional processing and is less extractive of cell contents, as a rule.
Cell layers usually require less time than more bulky samples, but even so I would expect that you will save time using MW processing. A disadvantage is that MW requires the user to be there pretty much constantly, since the steps are so short that it's difficult to wander off and do other things while your specimens sit in solutions for 15 minutes to an hour. Another interesting effect is that you may notice less apparent contrast in your stained grids, probably due to less extraction of cellular constituents leading to more even staining across structures (i.e., staining still works fine, but there is more to stain, leading to less apparent difference between adjacent components.)
Expense can be an issue, since a programmable laboratory microwave with variable wattage (almost a necessity these days), a vacuum chamber (highly recommended), a circulating water bath to prevent hot spots (really highly recommended) can set you back over $10,000US (about 8000 euros or so?). One could do MW processing with an ordinary kitchen microwave, using water and ice loads and neon bulbs to track hot spots, but that is, like, so last century. Not to mention far less repeatable and very inconvenient.
My feeling is that once you go MW, you will not go back, at least for the majority of your samples.
Hope this helps. Feel free to contact me if you have questions.
All the best, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Thursday, July 06, 2006 4:51 AM To: Tindall, Randy D.
Dear listers,
I am seriously considering investing in a microwave for our cell monolayers processing for TEM. I would like to have some comments from people already using this technique (advantages/disadvantages, worth the investment...) and also an idea of the money I would have to spend for it.
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Does anyone know where I can obtain an older model Edwards Pirani gauge part number PRCT10-P?
Thank you in advance,
Mick Thomas
------------------------------------------------- Mick Thomas UHV-STEM / FESEM Laboratory E-1 and F-3 Clark Hall (Lab) 212 Clark Hall (mail) Cornell University Ithaca, NY 14853
This is a very useful little program ... But more useful if it also included absrption edges.
genuinely :o) michael shaffer
SEM/MLA Research Coordinator Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
} All: the URL I previous sent has no spaces in it. Must have } been a finger mis-poke. } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTable/ } } r-holdford-at-ti.com wrote: } } } ---------------------------------------------------------------------- } } ------ The Microscopy ListServer -- CoSponsor: The } Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } } ------ } } } } Debby and anyone else who wants it: here's a link to } download the files: } } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } able/ } } } } Debby Sherman wrote: } } } } } Gee..would you mind sharing that program with others? I } would love } } } to have a copy. } } } } } } Debby } } } } } } Debby Sherman, Manager Phone: 765-494-6666 } } } Life Science Microscopy Facility FAX: 765-494-5896 } } } Purdue University E-mail: dsherman-at-purdue.edu } } } S-052 Whistler Building } } } 170 S. University Street } } } West Lafayette, IN 47907 } } } http://www.agriculture.purdue.edu/microscopy } } } } } } } } } On 6/28/06 6:37 PM, "r-holdford-at-ti.com" {r-holdford-at-ti.com} wrote: } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } } -------- The Microscopy ListServer -- CoSponsor: The Microscopy } } } } Society of America To Subscribe/Unsubscribe -- } } } } http://www.microscopy.com/MicroscopyListserver } } } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------------- } } } } -------- } } } } } } } } Owen: one of my favorites is EM Periodic Table, a } program authored } } } } by Scott Walck many years ago. It has a periodic chart } layout and a } } } } table layout that can be translated to Excel. I got it } at a Lehigh } } } } short course and I find it very handy. I'd be happy to } share it with you. } } } } Scott calls this "beerware" meaning when we meet him at } conferences, } } } } we owe him a beer if we use it. } } } } } } } } Scott: you can claim your pitcher at M&M in Chicago. } This program } } } } would be cheap at twice the price. } } } } } } } } opmills-at-mtu.edu wrote: } } } } } } } } } } } } } } ------------------------------------------------------------------- } } } } } --------- The Microscopy ListServer -- CoSponsor: The } Microscopy } } } } } Society of America To Subscribe/Unsubscribe -- } } } } } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help
==============================Original Headers============================== 5, 20 -- From Michael-at-Shaffer.net Thu Jul 6 12:07:14 2006 5, 20 -- Received: from ws6-1.us4.outblaze.com (ws6-1.us4.outblaze.com [205.158.62.196]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k66H7D2Q010236 5, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 12:07:13 -0500 5, 20 -- Received: (qmail 27077 invoked from network); 6 Jul 2006 17:07:09 -0000 5, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 5, 20 -- by ws6-1.us4.outblaze.com with SMTP; 6 Jul 2006 17:07:08 -0000 5, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 5, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 20 -- Subject: RE: [Microscopy] Re: need x-ray emission table 5, 20 -- Date: Thu, 6 Jul 2006 14:37:07 -0230 5, 20 -- Message-ID: {000a01c6a11e$9d7e0390$8d829986-at-roamingwolf} 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="us-ascii" 5, 20 -- Content-Transfer-Encoding: 7bit 5, 20 -- X-Mailer: Microsoft Office Outlook 11 5, 20 -- In-Reply-To: {200607051950.k65JonqE021573-at-ns.microscopy.com} 5, 20 -- Thread-Index: AcagbFPUO3EWLVX2ScWRsy+z/ZarYgAsfvoQ 5, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
Michael, The program does have the absorption edges in it. That's what an EELS line corresponds to. Check out Si for example. You will find the K-a line for Si at 1.74 in the XEDS mode. Then go to EELS mode and click on Si again and you will see that the Si K edge is 1839 eV.
BTW, to address the original question, the data files that come with the program are CSV (comma separated values) files and will open directly into Excel or another spreadsheet program. Two are XEDS data, one in keV and one it eV. The other file is the EELS data only and is only in eV. Before you copy it to use it, please look at the credits to Nestor, Noran, and EmiSpec for the data in the help menu of the program.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: michael-at-Shaffer.net [mailto:michael-at-Shaffer.net] Sent: Thursday, July 06, 2006 10:12 AM To: Walck-at-SouthBayTech.com
This is a very useful little program ... But more useful if it also included absrption edges.
genuinely :o) michael shaffer
SEM/MLA Research Coordinator Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
} All: the URL I previous sent has no spaces in it. Must have } been a finger mis-poke. } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicTab le/ } } r-holdford-at-ti.com wrote: } } } ---------------------------------------------------------------------- } } ------ The Microscopy ListServer -- CoSponsor: The } Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } } ------ } } } } Debby and anyone else who wants it: here's a link to } download the files: } } } http://www.amc.anl.gov/ANLSoftwareLibrary/EMMPDL(old)/Eels/EMPeriodicT } } able/ } } } } Debby Sherman wrote: } } } } } Gee..would you mind sharing that program with others? I } would love } } } to have a copy. } } } } } } Debby } } } } } } Debby Sherman, Manager Phone: 765-494-6666 } } } Life Science Microscopy Facility FAX: 765-494-5896 } } } Purdue University E-mail: dsherman-at-purdue.edu } } } S-052 Whistler Building } } } 170 S. University Street } } } West Lafayette, IN 47907 } } } http://www.agriculture.purdue.edu/microscopy } } } } } } } } } On 6/28/06 6:37 PM, "r-holdford-at-ti.com" {r-holdford-at-ti.com} wrote: } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } } -------- The Microscopy ListServer -- CoSponsor: The Microscopy } } } } Society of America To Subscribe/Unsubscribe -- } } } } http://www.microscopy.com/MicroscopyListserver } } } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------------- } } } } -------- } } } } } } } } Owen: one of my favorites is EM Periodic Table, a } program authored } } } } by Scott Walck many years ago. It has a periodic chart } layout and a } } } } table layout that can be translated to Excel. I got it } at a Lehigh } } } } short course and I find it very handy. I'd be happy to } share it with you. } } } } Scott calls this "beerware" meaning when we meet him at } conferences, } } } } we owe him a beer if we use it. } } } } } } } } Scott: you can claim your pitcher at M&M in Chicago. } This program } } } } would be cheap at twice the price. } } } } } } } } opmills-at-mtu.edu wrote: } } } } } } } } } } } } } } ------------------------------------------------------------------- } } } } } --------- The Microscopy ListServer -- CoSponsor: The } Microscopy } } } } } Society of America To Subscribe/Unsubscribe -- } } } } } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help
==============================Original Headers============================== 5, 20 -- From Michael-at-Shaffer.net Thu Jul 6 12:07:14 2006 5, 20 -- Received: from ws6-1.us4.outblaze.com (ws6-1.us4.outblaze.com [205.158.62.196]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k66H7D2Q010236 5, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 12:07:13 -0500 5, 20 -- Received: (qmail 27077 invoked from network); 6 Jul 2006 17:07:09 -0000 5, 20 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 5, 20 -- by ws6-1.us4.outblaze.com with SMTP; 6 Jul 2006 17:07:08 -0000 5, 20 -- From: "michael shaffer" {michael-at-Shaffer.net} 5, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 20 -- Subject: RE: [Microscopy] Re: need x-ray emission table 5, 20 -- Date: Thu, 6 Jul 2006 14:37:07 -0230 5, 20 -- Message-ID: {000a01c6a11e$9d7e0390$8d829986-at-roamingwolf} 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="us-ascii" 5, 20 -- Content-Transfer-Encoding: 7bit 5, 20 -- X-Mailer: Microsoft Office Outlook 11 5, 20 -- In-Reply-To: {200607051950.k65JonqE021573-at-ns.microscopy.com} 5, 20 -- Thread-Index: AcagbFPUO3EWLVX2ScWRsy+z/ZarYgAsfvoQ 5, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
==============================Original Headers============================== 16, 24 -- From walck-at-southbaytech.com Thu Jul 6 13:27:01 2006 16, 24 -- Received: from ylpvm01.prodigy.net (ylpvm01-ext.prodigy.net [207.115.57.32]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k66IR0fZ021802 16, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 13:27:00 -0500 16, 24 -- X-ORBL: [64.169.193.90] 16, 24 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 16, 24 -- by ylpvm01.prodigy.net (8.13.7 out spool5000 dk/8.13.7) with ESMTP id k66IQuY5000387; 16, 24 -- Thu, 6 Jul 2006 14:26:58 -0400 16, 24 -- From: "Scott Walck" {walck-at-southbaytech.com} 16, 24 -- To: {michael-at-Shaffer.net} 16, 24 -- Cc: {Microscopy-at-microscopy.com} 16, 24 -- Subject: RE: [Microscopy] need x-ray emission table 16, 24 -- Date: Thu, 6 Jul 2006 11:27:46 -0700 16, 24 -- Message-ID: {003201c6a129$e232a9e0$7801a8c0-at-dynamicbl8uno3} 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-Type: text/plain; 16, 24 -- charset="us-ascii" 16, 24 -- Content-Transfer-Encoding: 7bit 16, 24 -- X-Priority: 3 (Normal) 16, 24 -- X-MSMail-Priority: Normal 16, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 16, 24 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 16, 24 -- Importance: Normal 16, 24 -- In-Reply-To: {200607061712.k66HCE9x016726-at-ns.microscopy.com} ==============================End of - Headers==============================
I am looking for a side mounted camera assembly for a FEI Tecnai 12. Philips CM or 400 series TEM. While a complete assembly, including camera, would be ideal, I have some digital cameras I can mount if I can get the ports and the yag/prism sidearm assembly.
Please contact me offline in this regard if you have a unit you are willing to part with
Steve
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu
==============================Original Headers============================== 6, 18 -- From sbarlow-at-sunstroke.sdsu.edu Thu Jul 6 14:32:17 2006 6, 18 -- Received: from sciences.sdsu.edu (sciences.sdsu.edu [130.191.140.33]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k66JWH49000947 6, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 14:32:17 -0500 6, 18 -- Received: from [146.244.225.122] ([146.244.225.122]) 6, 18 -- by sciences.sdsu.edu (8.12.11.20060614/8.12.10/SCEC-032005-9-50) with ESMTP id k66JWFDp026671 6, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 12:32:16 -0700 (PDT) 6, 18 -- Mime-Version: 1.0 6, 18 -- Message-Id: {p06110405c0d3154fc4d8-at-[146.244.225.122]} 6, 18 -- Date: Thu, 6 Jul 2006 12:32:08 -0700 6, 18 -- To: Microscopy-at-microscopy.com 6, 18 -- From: Steve Barlow {sbarlow-at-sunstroke.sdsu.edu} 6, 18 -- Subject: (TEM) used side mounted camera assembly 6, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 18 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-1.6 (sciences.sdsu.edu [130.191.140.35]); Thu, 06 Jul 2006 12:32:16 -0700 (PDT) 6, 18 -- X-COS-MailScanner: Found to be clean 6, 18 -- X-MailScanner-SpamCheck: not spam, SpamAssassin (score=0, required 8) 6, 18 -- X-MailScanner-From: sbarlow-at-sunstroke.sdsu.edu ==============================End of - Headers==============================
Listers, I am trying to fix an old LBK IV ultramicrotome Model 2128. The metal band that connects the drive motor to the specimen arm has broken. I can probably carry out the repairs myself but need to locate a replacement part. I think that there is no official parts source these days but wondered if anyone had already cannibalized such an instrument or had an unused one suitable for spares. Any help much appreciated.
Best regards
Chris
Christopher J Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.A04 UT Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-9039 Phone +1 214 648 2827 Fax +1 214 648 6408
==============================Original Headers============================== 6, 21 -- From christopher.gilpin-at-utsouthwestern.edu Thu Jul 6 17:43:51 2006 6, 21 -- Received: from swlx166.swmed.edu (swlx166.swmed.edu [199.165.152.166]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k66MhpKo014100 6, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 6 Jul 2006 17:43:51 -0500 6, 21 -- Received: from [129.112.148.58] (helo=cgdesktop) 6, 21 -- by swlx166.swmed.edu with esmtp (Exim 4.44) 6, 21 -- id 1FycZZ-0002YK-Kh 6, 21 -- for Microscopy-at-microscopy.com; Thu, 06 Jul 2006 17:43:51 -0500 6, 21 -- From: "Christopher Gilpin" {christopher.gilpin-at-utsouthwestern.edu} 6, 21 -- To: {Microscopy-at-microscopy.com} 6, 21 -- Date: Thu, 6 Jul 2006 17:43:58 -0500 6, 21 -- Message-ID: {001301c6a14d$ab4d2030$3a947081-at-cgdesktop} 6, 21 -- MIME-Version: 1.0 6, 21 -- Content-Type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-Transfer-Encoding: 7bit 6, 21 -- X-Mailer: Microsoft Office Outlook 11 6, 21 -- Thread-Index: AcahTasR/3FlEY2zSBiQQoQxao6KCA== 6, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 6, 21 -- X-Scan-Signature: caec9e7ca10ae3625b33f2c31db97675 6, 21 -- Subject: Parts and repair for LKB IV ultramicrotome ==============================End of - Headers==============================
It gets pretty hot near the filament. I suspect that the sputtered on coating would flake off due to the differential thermal expansion of the two metals since conventionally sputtered metals are sitting lightly on the surface. On the other hand, electroplated metals would be more stable. For example, we have an ancient Wehnelt from an RCA that is gold plated. It was stable for 30 or more years. BUT, then, apparently, someone tried to clean it too vigorously and removed some of the gold. Now the cleaned area is tarnished.
Perhaps Steve Chapman and Chuck Garber might want to chime in here as they are experienced in scope maintenance and metalurgy, respectively.
JB -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
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Email: shapiro-at-unbc.ca Name: Aaron Shapiro
Organization: University of Northern British Columbia
Title-Subject: [Filtered] Human CNS Atlas
Question: I was wondering if anyone was aware of a good histology atlas which shows TEM images of human or rat central nervous system - particularly the granule cell layer of the cerebellum.
you can also try MA-Table for X-ray emission lines, wisible with EDS. There are more lines in L- and M-series than in other data basis. This is the manual and there you'll find also the download: http://microanalyst.mikroanalytik.de/manual.html
Regards
Frank
opmills-at-mtu.edu schrieb:
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==============================Original Headers============================== 7, 19 -- From eggert-at-mikroanalytik.de Fri Jul 7 00:57:32 2006 7, 19 -- Received: from Mail1.KONTENT.De (mail1.kontent.de [81.88.34.36]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k675vW2f026780 7, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 00:57:32 -0500 7, 19 -- Received: from mikroanalytik.de (p54BDD829.dip.t-dialin.net [84.189.216.41]) 7, 19 -- by Mail1.KONTENT.De (Postfix) with ESMTP id 290C3108FAFB; 7, 19 -- Fri, 7 Jul 2006 07:57:33 +0200 (CEST) 7, 19 -- Message-ID: {44ADF8B4.7090102-at-mikroanalytik.de} 7, 19 -- Date: Fri, 07 Jul 2006 08:01:24 +0200 7, 19 -- From: Frank Eggert {eggert-at-mikroanalytik.de} 7, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; de-DE; rv:1.4) Gecko/20030619 Netscape/7.1 (ax) 7, 19 -- X-Accept-Language: de, de-de, en 7, 19 -- MIME-Version: 1.0 7, 19 -- To: opmills-at-mtu.edu, Microscopy-at-microscopy.com 7, 19 -- Subject: Re: [Microscopy] need x-ray emission table 7, 19 -- References: {200606282002.k5SK2YYY024728-at-ns.microscopy.com} 7, 19 -- In-Reply-To: {200606282002.k5SK2YYY024728-at-ns.microscopy.com} 7, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
We are happy to announce the first release of the 2dx software.
2dx provides a graphical user interface for the computer processing of 2D crystal images. So far, the program 2dx_image is available for the processing of individual images of tilted or non-tilted 2D crystals. Merging will be implemented in the near future.
2dx assists running c-shell scripts that call other stand-alone programs. In the current release we provide a set of MRC programs, which were slightly adapted to interact with the 2dx software, and we added a few functions trying to implement automatic determination of all required parameters. It is easy to adapt or replace the provided scripts with your own scripts and/or to make use of other back-end programs (Spider, bsoft, etc.) or your own MRC software.
2dx including the source codes is freely available, and we provide compiled versions for Linux x86_64bit and Mac-PPC (G4 (slow), G5) and the new Mac-Intel computers, see here: http://2dx.org The installer hopefully take care of everything (all goes into /usr/ local/2dx), and installs also the required modified MRC programs. The only thing that is required in addition is CCP4, as described on the server. But CCP4 is only used in the last script for the generation of a map. The rest should also work without CCP4. On the server are also some test data, in MRC-image2000 file format for big and small endian machines, together with the corresponding config files (2dx_image.cfg).
A manuscript that describes this software is under review at JSB, but we don't have written the manual for 2dx yet. However, hopefully, most features in 2dx are self-explanatory. Just try the right mouse button over all the different items, or double-clicking.
Bryant, Xiangyan and Henning.
Henning Stahlberg, Molecular & Cellular Biology, Briggs Hall 5, University of California at Davis, 1 Shields Ave., Davis, CA 95616, USA Tel: +1-530-752 8282 (office), +1-530-754 8285 (lab) Fax: +1-530-752 3085 mailto:HStahlberg-at-ucdavis.edu http://stahlberglab.ucdavis.edu AIM:HStahlberg-at-mac.com
Workshop on Electron Crystallography, August 7-11, 2006, at UC Davis. http://2dx.org/workshop
2dx Software Version 1.0.0 now Available http://2dx.org/download/2dx-software/ _____________________________________________________________
==============================Original Headers============================== 18, 16 -- From HStahlberg-at-ucdavis.edu Fri Jul 7 15:40:07 2006 18, 16 -- Received: from pop23.ucdavis.edu (pop23.ucdavis.edu [169.237.105.13]) 18, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k67Ke7ol026329 18, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 15:40:07 -0500 18, 16 -- Received: from [169.237.214.65] ([169.237.214.65]) 18, 16 -- by pop23.ucdavis.edu (8.13.6/8.13.1/it-std-5.2.0) with ESMTP id k67Ke7a7014362 18, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 13:40:07 -0700 (PDT) 18, 16 -- Mime-Version: 1.0 (Apple Message framework v752.2) 18, 16 -- Content-Transfer-Encoding: 7bit 18, 16 -- Message-Id: {EACF3DDD-9442-4698-98F9-9B64934B101C-at-ucdavis.edu} 18, 16 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 18, 16 -- To: Microscopy-at-microscopy.com 18, 16 -- From: Henning Stahlberg {HStahlberg-at-ucdavis.edu} 18, 16 -- Subject: 2dx - image processing for 2D membrane protein crystals 18, 16 -- Date: Fri, 7 Jul 2006 13:40:01 -0700 18, 16 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
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I have a question regarding image-diffraction pattern rotation in TEM. While in some TEMs this rotation is compensated, it is still possible to have a 180deg rotation between a TEM image and the corresponding diffraction pattern. What are the procedures to measure this rotation with or without special calibration samples?
I have a user who wants to know how thick a metal coating has to be to clog up the pores in a nucleopore filter.
I know, why not just try different amounts until it works, but this is an engineer type who wants to know the real numbers.
For this project he is trying to restrict the number of open pores in the filter to just a few in the center of his 1 cm dia. filter. The plan to do this for now is to put a 1 mm x 1 mm mask on the filter and evaporate or sputter some metal over the rest.
We were thinking gold or gold/paladium, but if you have a better idea, we're game for that.
The pores he wants to clog are 10 nm, 30 nm, 50 nm or 80 nm in a standard polycarbonate nucleopore filter. Objective is to stop fluid from going through the clogged pores, while still keeping the central 1 mm square pores open. The clogging stuff does not have to be electrically conductive, metal coating was just our first guess at a way to do it. Other strategies OK if they will work.
Thanks
Jon
--
Jonathan Krupp Microscopy & Imaging Lab C230 Earth & Marine Science University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-ucsc.edu
==============================Original Headers============================== 12, 21 -- From jmkrupp-at-ucsc.edu Fri Jul 7 17:38:22 2006 12, 21 -- Received: from smtp-prod-mx1.ucsc.edu (smtp-prod-mx1.ucsc.edu [128.114.125.43]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k67McLFb015788 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 17:38:21 -0500 12, 21 -- Received: from ucsc.edu (cruzmail-fe1.ucsc.edu [128.114.125.5]) 12, 21 -- by smtp-prod-mx1.ucsc.edu (8.13.7/8.13.7) with ESMTP id k67McH9r018876 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 15:38:17 -0700 (PDT) 12, 21 -- Received: from [128.114.25.141] (account jmkrupp-at-ucsc.edu [128.114.25.141] verified) 12, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 12, 21 -- with ESMTPA id 62079501 for microscopy-at-microscopy.com; Fri, 07 Jul 2006 15:38:17 -0700 12, 21 -- Mime-Version: 1.0 12, 21 -- Message-Id: {p06230906c0d4903dcf6c-at-[128.114.25.141]} 12, 21 -- Date: Fri, 7 Jul 2006 15:38:16 -0700 12, 21 -- To: microscopy-at-microscopy.com 12, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 12, 21 -- Subject: Thickness of metal coating needed 12, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 12, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 12, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 12, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 12, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
Go to convergent beam mode and go to cross over with the condenser lens. Then decrease the lens strength on the condenser (CCW) until you see the shadow image of the sample in the bright field disk. It should agree with the image orientation in the mag mode. (I think all microscopes have the knobs go clockwise to increase lens strength and CCW to decrease lens strength on the lenses.)
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: gradecak-at-fas.harvard.edu [mailto:gradecak-at-fas.harvard.edu] Sent: Friday, July 07, 2006 2:54 PM To: Walck-at-SouthBayTech.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both gradecak-at-fas.harvard.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
I have a question regarding image-diffraction pattern rotation in TEM. While in some TEMs this rotation is compensated, it is still possible to have a 180deg rotation between a TEM image and the corresponding diffraction pattern. What are the procedures to measure this rotation with or without special calibration samples?
We recently are having serious embedding problems using EPON generic (LX-112) formulation that we have used for years. Most of the problems seem to be in high pressure frozen samples including cultured mammalian cells and single cell cyanobacteria. Both samples embedded just fine a few months ago as did plant samples in the same resin formulation. Current cells are cryo-protected with dextran (~10% final concentration) and freezing looks great.
Cells themselves are well infiltrated but problems result in sections not spreading well and resin pulling away from cell walls (cyanobacteria). The samples go through a very long (5day) gradual infiltration, gentle agitation, and attention is paid to keeping everything dry, etc...all the things we have been doing for years with never a problem.
Any suggestions would be appreciated. I am about to revert back to Spurr resin even though we may have problems with the new VCD replacement. We went to the EPON formulation to get better contrast. Suggestions as to other resins or resin mixtures would also be appreciated.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 7, 21 -- From dsherman-at-purdue.edu Fri Jul 7 21:50:59 2006 7, 21 -- Received: from 1061exfe03.adpc.purdue.edu (1061exfe03.adpc.purdue.edu [128.210.63.225]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k682ox3o006266 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Jul 2006 21:50:59 -0500 7, 21 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe03.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 21 -- Fri, 7 Jul 2006 22:50:59 -0400 7, 21 -- Received: from 74.132.211.81 ([74.132.211.81]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 7, 21 -- Sat, 8 Jul 2006 02:50:56 +0000 7, 21 -- User-Agent: Microsoft-Entourage/11.2.4.060510 7, 21 -- Date: Fri, 07 Jul 2006 22:50:55 -0400 7, 21 -- Subject: Embedding problems 7, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 7, 21 -- Message-ID: {C0D495CF.3EC2%dsherman-at-purdue.edu} 7, 21 -- Thread-Topic: Embedding problems 7, 21 -- Thread-Index: AcaiOVUHk4GIvg4sEdu0zgAKlcoUxg== 7, 21 -- Mime-version: 1.0 7, 21 -- Content-type: text/plain; 7, 21 -- charset="US-ASCII" 7, 21 -- Content-transfer-encoding: 7bit 7, 21 -- X-OriginalArrivalTime: 08 Jul 2006 02:50:59.0628 (UTC) FILETIME=[57C93EC0:01C6A239] ==============================End of - Headers==============================
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Email: mdyousuf-at-qu.edu.qa Name: Mohammed Yousuf
Organization: Qatar University
Title-Subject: [Filtered] Philips CM12 - digital cameras
Question: Dear All, Please advice regarding purchase of a new digital camera for our CM12 scope. As of now the scope is in average health given the time that it has served since its instillation in 1992. My question is - is it adviseable to go in for a digital camera to this instrument? If yes, what are the options available out there. I need information from places who have had a recent instillation. Also need to know the price/ performance index for various options (individual brands/models and their resolution). Kindly respond to me direct.
Thanks in advance.
Mohammed Yousuf EM unit, Central Research Laboratories Qatar University P.O.Box 2713 Doha, State of Qatar mdyousuf-at-qu.edu.qa mdyousuf99-at-yahoo.com
This Question was submitted to Ask-A-Microscopist by (lasvegassierra-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, July 9, 2006 at 07:52:01 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lasvegassierra-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I found this site kinda by chance. I am excited at the possiblility of sharing some findings of mine. I also hope that maybe one of the scientist can help me to identify this thing. I dont know if anyone is willing to try or how I would send you a copy. Thru regular email is one thing but this program is different. Is it possible for you to view my microscopic views with me? Thank you for your time. Amie
I am a bit puzzled by your questions because you used methanol on other materials and it appeared to work for you. Anyway, the questions you asked are probably of general use. You asked: 1) does methanol evaporation cause particle agglomeration on the grid? I think you mean flocculation and not agglomeration. Yes, pure methanol does cause this problem and so do other solvents. It depends on the nature of your particles as far as the materials they are made up of and whether they are hydrophobic or hydrophilic as well. These are not the only variables by a long shot.
} 2) does methanol use cause morphological changes of the particles? Not to the primary particles or primary structural units, unless they dissolve in MeOH.
} 3) are particles size-fractionated on the TEM grid during the } evaporation process? Absolutely. Particles do not 'drop out' of solution by gravity onto a grid. The particles move around in a dynamic way. So they can be segregated. Some of this can be stopped to get good representative dispersions. Furthermore, dropping the particle suspension onto a grid on a piece of filter paper should be avoided unless you want to skew the size results to get a biased result. This is done in some patents and is a 'hidden' bias in one ASTM TEM method on carbon blacks. Flocculation on a grid can also combine aggregate structural units to make you think the 'particle sizes' are larger.
} 4) does methanol dissolve some phases? I guess the technical answer is yes, if the phase is even slightly soluble in methanol. I would not think this applies to your 'flyash' samples that should have things like glassy particles and unburned coal or carbon in them.
} My question to the Listserver is - should I avoid methanol? Not if you need a slightly polar solvent to disperse the particles of interest. } which solvents should I use and why? Try different solvent systems. Use the one you think works the best for your sample situation.
General comments: Don't get tunnel vision when dispersing powders, aggregates, or primary particles. Try different solvents and look at them in a TEM. Look at how they behave. Examine the whole grid to see the different patterns you see as a dried dispersion.
There's no magic bullet single dispersing system here. That's certainly true of colored ink jet pigment dispersions. The number of factors that come into play in dispersing powders is not a single dimensional variable called a solvent. Here's a pointed example of tunnel vision. I was told that a sample of some plasma generated nanoparticles was hydrophobic. The newly hired chemist's sonicated settling tests showed it would not disperse in water based systems. What would you use? I was told which organic solvent to use and I used that one. Then I dispersed the hydrophobic sample in a water based system. Both preps gave equivalent flocculation free dispersions on TEM grids. This was done by manipulating the other factors important in dispersion besides the type of solvent system.
In a previous posting from Leslie at IBM the following summary item was detailed. } There were two (who) mentioned dry loading the grid, which is dumping some of } the powder on the grid and tapping off the excess. This is the method I } tried first as it was the easiest, but I have not looked at the grid yet } to see if there is enough material to do EELS on. This (DeGussa-Germany?) method relies on the 'adhesion' of the finest aggregates to a grid film. They are attached simply by static charge. My experience with silica and plasma nanoparticles was that the populations were usually too light for me to use. The aggregate size distribution was skewed towards smaller sizes. Another problem is that the powder aggregates get on both sides of the grid without special handling precautions. This causes some aggregates on the wrong side of the grid, to be over focused. Yet, dry preps are faster to perform than solvent preps or mulling. I used dry preps when speed was critical, and the structure or size was not of interest.
Recently, the change in the size of ppt'd silica particles was blamed on a certain dispersing solvent. A dry prep would have shown that the size increased without that solvent ever being used. A dry prep is a good referee technique. For me, high speed high-res imaging of silica dry preps showed the presence of single digit nanometer scale salt on the surfaces of primary particles that accelerates this above mentioned "poached egg" look but it would happen even without the trace salts, given enough beam time. So dry preps have a place in a TEM lab and are useful.
Disclaimers: I performed dispersions on all types of powders and examined thin sections of the interiors of agglomerates, large aggregates, coatings of all types, and silica tire cleats for over 20 years. Your samples might perform differently than stated here. There are at least 10 factors or variables that apply to dispersions of powders and aggregates. That's too many to discuss here. Obtaining a good dispersion is a learned skill and takes practice.
Paul Beauregard Senior Research Associate Greensburg, PA 724-834-2247
At 09:10 PM 6/25/06 -0500, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 13, 24 -- From beaurega-at-westol.com Mon Jul 10 08:54:47 2006 13, 24 -- Received: from smtp-gateway-5.winbeam.com (smtp-gateway-5.winbeam.com [64.84.97.70]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6ADskA1016864 13, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Jul 2006 08:54:47 -0500 13, 24 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 13, 24 -- by smtp-gateway-5.winbeam.com (8.13.2/8.12.8) with SMTP id k6ADsTX9028600 13, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 10 Jul 2006 09:54:29 -0400 13, 24 -- Received: (qmail 26830 invoked by uid 89); 10 Jul 2006 13:54:27 -0000 13, 24 -- Received: from pitts-69-72-110-171.dynamic-dialup.coretel.net (HELO millenium) (69.72.110.171) 13, 24 -- by mail.winbeam.com with SMTP; 10 Jul 2006 13:54:27 -0000 13, 24 -- Message-Id: {3.0.6.32.20060710095550.007f8560-at-pop3.norton.antivirus} 13, 24 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus (Unverified) 13, 24 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 13, 24 -- Date: Mon, 10 Jul 2006 09:55:50 -0400 13, 24 -- To: mgb-at-ansto.gov.au, Microscopy-at-microscopy.com, lkrupp-at-us.ibm.com 13, 24 -- From: Beaurega {beaurega-at-westol.com} 13, 24 -- Subject: Re: [Microscopy] TEM powder sample preparation 13, 24 -- Mime-Version: 1.0 13, 24 -- Content-Type: text/plain; charset="us-ascii" 13, 24 -- X-Winbeam-MailScanner-Information: - Please contact Technical Support for more information 13, 24 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) 13, 24 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, score=0, 13, 24 -- required 6, autolearn=disabled) 13, 24 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
One of our users is asking for references (books, articles etc.) concerning the accuracy of SEM measurements for particles. I explained to him the principles (difficulties with edge effect etc.), but he needs printed references for his article.
If anyone has an article name/book chapter, I would greatly appreciate it.
Thanks, Daniel Salamon Technical Officer, Electron Microscopy
National Institute for Nanotechnology, NRC 11421 Saskatchewan Dr. Edmonton, AB. T6G 2M9
Phone: (780) 641-1663 Fax: (780) 641-1601
==============================Original Headers============================== 7, 23 -- From Daniel.Salamon-at-nrc-cnrc.gc.ca Mon Jul 10 10:18:12 2006 7, 23 -- Received: from nrccenexf2.nrc.ca (nrccenexf2.nrc.ca [132.246.15.83]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6AFIBRM028823 7, 23 -- for {microscopy-at-microscopy.com} ; Mon, 10 Jul 2006 10:18:12 -0500 7, 23 -- Received: from nrccenexb2.nrc.ca ([132.246.15.98]) by nrccenexf2.nrc.ca with Microsoft SMTPSVC(6.0.3790.1830); 7, 23 -- Mon, 10 Jul 2006 11:18:06 -0400 7, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 23 -- Content-class: urn:content-classes:message 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-Type: text/plain; 7, 23 -- charset="us-ascii" 7, 23 -- Subject: SEM magnification accuracy references 7, 23 -- Date: Mon, 10 Jul 2006 11:18:05 -0400 7, 23 -- Message-ID: {923687253229634E96E808B8D3124C839789C3-at-nrccenexb2.nrc.ca} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: SEM magnification accuracy references 7, 23 -- Thread-Index: AcakNAquO+NWUArVRvi14ZEbnys6cQ== 7, 23 -- From: "Salamon, Daniel" {Daniel.Salamon-at-nrc-cnrc.gc.ca} 7, 23 -- To: {microscopy-at-microscopy.com} 7, 23 -- X-OriginalArrivalTime: 10 Jul 2006 15:18:06.0202 (UTC) FILETIME=[0B551DA0:01C6A434] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6AFIBRM028823 ==============================End of - Headers==============================
X-from cross over, under- or over-focusing the condenser lens will make both make it parallel. In the analytical machines that have more than two condenser lenses (as in the last 20+) years, you obtain the parallel condition faster (by less turns of the knob) by over-focusing the C2 lens than going under-focus. In machines that only have the two condenser system, you under focus the condenser to go to parallel condition faster. In either case, you never get to a parallel condition, you get to a very small convergence angle.
With respect to my previous answer, I believe that you can check the technique that I gave with a GaAs [011] CBED image since it is a non-centrosymmetric crystal.
As far as I know, TEM's still have the condition that CW and CCW still relate to the strength in the lens. You can watch the current or voltage monitor of the lenses when you turn the knob. Some manufacturer's of SEM's have changed that, especially with the objective lens. Now they relate the CW turn of the focus know with increasing the working distance, which of course, is decreasing the strength of the objective lens.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: Dusha [mailto:a.chuvilin-at-microscopist.ru] Sent: Sunday, July 09, 2006 8:36 AM To: walck-at-southbaytech.com
Don't to forget about dimensional calibration of the SEM. If this is not correct, all bets are off.
gary g.
At 08:20 AM 7/10/2006, you wrote:
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My first thought would be to use this filter exactly as he is intended to: to filter! Masking the central part, you can filter a suspension of particles which will eventually block the filter pores. I am confident that precipitates likes calcium carbonate or calcium oxalate would block the filter. Of course it all depends on the solution you have to filter (and its pH). Any particulate suspension will block the filter. Chemically neutral particles like fine mineral powder (silicate?) should work well too. It very much depends on what is available to you.
Regards,
Stephane
--- jmkrupp-at-ucsc.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi: } } I have a user who wants to know how thick a metal } coating has to be } to clog up the pores in a nucleopore filter. } } I know, why not just try different amounts until it } works, but this } is an engineer type who wants to know the real } numbers. } } For this project he is trying to restrict the number } of open pores in } the filter to just a few in the center of his 1 cm } dia. filter. The } plan to do this for now is to put a 1 mm x 1 mm mask } on the filter } and evaporate or sputter some metal over the rest. } } We were thinking gold or gold/paladium, but if you } have a better } idea, we're game for that. } } The pores he wants to clog are 10 nm, 30 nm, 50 nm } or 80 nm in a } standard polycarbonate nucleopore filter. Objective } is to stop fluid } from going through the clogged pores, while still } keeping the central } 1 mm square pores open. The clogging stuff does not } have to be } electrically conductive, metal coating was just our } first guess at a } way to do it. Other strategies OK if they will work. } } Thanks } } Jon } } } -- } } } Jonathan Krupp } Microscopy & Imaging Lab } C230 Earth & Marine Science } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-ucsc.edu } } ==============================Original } Headers============================== } 12, 21 -- From jmkrupp-at-ucsc.edu Fri Jul 7 17:38:22 } 2006 } 12, 21 -- Received: from smtp-prod-mx1.ucsc.edu } (smtp-prod-mx1.ucsc.edu [128.114.125.43]) } 12, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k67McLFb015788 } 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Jul 2006 17:38:21 -0500 } 12, 21 -- Received: from ucsc.edu } (cruzmail-fe1.ucsc.edu [128.114.125.5]) } 12, 21 -- by smtp-prod-mx1.ucsc.edu (8.13.7/8.13.7) } with ESMTP id k67McH9r018876 } 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 } Jul 2006 15:38:17 -0700 (PDT) } 12, 21 -- Received: from [128.114.25.141] (account } jmkrupp-at-ucsc.edu [128.114.25.141] verified) } 12, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP } 4.3.7) } 12, 21 -- with ESMTPA id 62079501 for } microscopy-at-microscopy.com; Fri, 07 Jul 2006 15:38:17 } -0700 } 12, 21 -- Mime-Version: 1.0 } 12, 21 -- Message-Id: } {p06230906c0d4903dcf6c-at-[128.114.25.141]} } 12, 21 -- Date: Fri, 7 Jul 2006 15:38:16 -0700 } 12, 21 -- To: microscopy-at-microscopy.com } 12, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} } 12, 21 -- Subject: Thickness of metal coating needed } 12, 21 -- Content-Type: text/plain; } charset="us-ascii" ; format="flowed" } 12, 21 -- X-UCSC-CATS-Information: This message was } scanned by the ITS MailScanner } 12, 21 -- X-UCSC-CATS-MailScanner: Found to be clean } 12, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: } 12, 21 -- X-UCSC-CATS-MailScanner-From: } jmkrupp-at-ucsc.edu } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Tue Jul 11 01:41:38 2006 9, 20 -- Received: from web37403.mail.mud.yahoo.com (web37403.mail.mud.yahoo.com [209.191.87.56]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6B6fcNL010706 9, 20 -- for {microscopy-at-microscopy.com} ; Tue, 11 Jul 2006 01:41:38 -0500 9, 20 -- Received: (qmail 15945 invoked by uid 60001); 11 Jul 2006 06:41:38 -0000 9, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 20 -- s=s1024; d=yahoo.com; 9, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 9, 20 -- b=1K8znUBb9IFipbyStKpnQyeM1FsMVyXD0bwftsmJjKQJVkto7Fj94hKsZB/mvEn4qn/zVWNlo1e4HGl/mOwueLj8XMVgt7sfOD3qv7+7fPUMZxXHoGAvUIM6Y+t2X0kHKaIwfGaREg9ShXGabEXLHGvbMeqpxs5s2/EOeE5NoU8= ; 9, 20 -- Message-ID: {20060711064138.15943.qmail-at-web37403.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37403.mail.mud.yahoo.com via HTTP; Mon, 10 Jul 2006 23:41:38 PDT 9, 20 -- Date: Mon, 10 Jul 2006 23:41:38 -0700 (PDT) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] Thickness of metal coating needed 9, 20 -- To: jmkrupp-at-ucsc.edu 9, 20 -- Cc: m